Neoplasma

EGFR/STAT3 signaling mediates the upregulation of CD47 in HPV-positive cervical cancer by activating p65 and exosome transporter RAB31

In cervical cancer, the regulatory mechanisms of CD47 and its enrichment in exosomes have not been fully elucidated. In this study, we aim to explore the mechanisms of how EGFR/STAT3 signaling regulates CD47 expression in cervical cancer, and whether p65 or RAB31 is involved in this process. RNA interference was used to knock down the fragment of HPV in cervical cancer cells. EGFR and RAB31 phosphorylation were detected by western blot, and exosomal CD47 was determined by ELISA. EGFR or STAT3 was then knocked down and western blot was used to detect the expression or phosphorylation of EGFR, STAT3, p65, CD47, RAB31, and its related proteins. ChIP-qPCR was used to investigate p65 enrichment in the CD47 promoter region. Our results showed that HPV fragment knockdown reduced the phosphorylation of EGFR and RAB31, and exosomal CD47 expression. Knocking down EGFR inhibited phosphorylation of STAT3 and p65, and expression of RAB31-related proteins. Phosphorylated p65 was bound to the P3 and P7 promoter regions of CD47. EGFR/STAT3 signaling upregulated CD47 expression by phosphorylating p65 and enhanced exosomal CD47 by activated RAB31. Thus, a dual regulatory role of EGFR/STAT3 signaling on CD47 expression and secretion involving transcriptional factor p65 and exosome transporter RAB31 was demonstrated.

Hypoxia-induced miR-9 expression promotes ovarian cancer progression via activating PI3K/AKT/mTOR/GSK3β signaling pathway

Ovarian cancer (OC) is one of the most prevalent malignant tumors affecting women's life and health. Since OC has a poor prognosis due to extensive metastasis, there is a need to explore a new mechanism of OC metastasis. microRNAs (miRs) are single-stranded, non-coding RNAs. miR-9 has been reported to promote cancer and may provide a new strategy for OC diagnosis. The purpose of this study was to examine the function and underlying mechanism of miR-9 in OC. RT-qPCR was used to assess miR-9 expression levels. Transwell assays were used to determine the number of migrating and invading OC cells. The protein expression levels of the PI3K/AKT/mTOR/GSK3β signaling pathway were examined using western blotting. The results informed that, when compared to normal ovarian tissues, miR-9 was remarkably expressed in OC tissues, and hypoxia might lead to overexpression of miR-9-5p while inhibiting miR-9 notably suppressed the migrating and invading cell numbers in OC cells. In vivo, miR-9-5p knockdown inhibited tumor growth in a subcutaneous nude mice model of SKOV3 cells. Our findings suggest that miR-9 could be an underlying oncogene in OC, opening up new avenues for OC diagnosis and treatment of OC by targeting miR-9.

TRIM11 regulated by m6A modification promotes the progression of cervical cancer by PHLPP1 ubiquitination

Cervical cancer (CC) is a common cancer in women and a serious threat to women's lives. TRIM11 has been confirmed as a carcinogen in multiple cancers. Here, we will excavate the detailed mechanism of TRIM11 in CC. CC cell lines and nude mice were experimental subjects in this study. The abundance of genes and proteins was detected using qRT-PCR, western blot, and IHC. Cell proliferation, migration, and invasion were determined by CCK-8 assay, wound healing assay, and Transwell, respectively. The interactions among METTL14, TRIM11, and PHLPP1 were confirmed using RIP and co-IP, respectively. The stability of TRIM11 mRNA was examined by qRT-PCR with actinomycin D treatment. The m6A level of TRIM11 was detected by MeRIP assay. Results showed that TRIM11 levels were elevated in CC cells. TRIM11 depletion attenuated the proliferation, migration, and invasion of Hela and SiHa cells. Additionally, TRIM11 was modified with m6A, which was mediated by METTL14, and the stability of TRIM11 mRNA was enhanced by IGF2BP1 depending on the level of m6A modification. TRIM11 ubiquitinated PHLPP1 and led to reduced PHLPP1 expression at the protein level. PHLPP1 could further result in the dephosphorylation of AKT and inhibit AKT signaling. PHLPP1 knockdown neutralized TRIM11 silencing-mediated repression of malignant phenotypes of CC cells. TRIM11 mediated by the METTL14-IGF2BP1 axis promotes the AKT pathway to accelerate CC progression by mediating the ubiquitination of PHLPP, which might provide novel therapeutic targets for CC treatment.

Xenogeneic fibroblasts inhibit the growth of the breast and ovarian cancer cell lines in co-culture

Cell-based therapies cure some hematologic malignancies, although little information exists on solid cancer cell responses. The study objective was to test the hypothesis that xenogeneic fibroblasts can inhibit the growth of human cancer cell lines in vitro. Seven human cell lines (pancreatic cancer HPAF II; brain cancer U-87 MG; fibrosarcoma; ovarian cancer OVCAR3 and SKOV3; and breast cancer MCF7 and MDA-MB231) were co-cultured with two xenogeneic fibroblast cell lines (CV-1; monkey, Cercopithecus aethiops and DF-1; chicken, Gallus gallus) in a Transwell culture system. Cancer cell proliferation was assessed colorimetrically. Different concentrations of breast and ovarian cancer cells were tested. Gene expression induced by DF-1 xenogeneic fibroblasts was assessed by RNAseq of MCF7 breast cancer cells. The proliferation of the majority of the cancer cell lines was altered by co-culture with xenogeneic fibroblasts. Cell proliferation was increased (4-17%) by CV-1; DF-1 increased brain cancer cell proliferation (16%), decreased breast and ovarian cancer cell growth (15 and 26% respectively) but did not affect fibrosarcoma and pancreatic cancer cells. When the initial cancer cell concentrations were lowered 4-fold, growth inhibition of breast and ovarian cancer increased more than 2-fold. DF-1 fibroblasts induced significant differential expression of 484 genes in MCF7 breast cancer cells; 285 genes were downregulated and 199 genes were upregulated compared to control. Genes involved in the immune response were the major downregulated entities. RNAseq results were validated by qRT-PCR of 12 genes. The results show that xenogeneic fibroblasts can alter the growth and gene expression of cancer cells in vitro. This suggests a potentially novel investigational approach to the control of cancer cell growth.

Circular RNA_0000326 accelerates breast cancer development via modulation of the miR-9-3p/YAP1 axis

Circular RNA (circ)_0000326 has been reported in bladder cancer and cervical cancer and is concerned to be involved with the development of cancerous cells. Whereas, there have been no reports concentrating on the influences of circ_0000326 in breast cancer (BC). Therefore, the latent modulatory mechanisms of circ_0000326 in BC are researched. circ_0000326 expression in BC tissues and correlative cells was evaluated via RT-qPCR, and the relevance between circ_0000326 expression and overall survival and the clinicopathological feature was also investigated. After a series of transfection, the effects of circ_0000326, microRNA-9-3p (miR-9-3p), and Yes-associated protein 1 (YAP1) in BC cell growth, invasion, and stemness were studied by CCK-8, flow cytometry, Transwell, and sphere-forming assays. The binding sites and correlation of circ_0000326, miR-9-3p, and YAP1 were certified via starBase website, luciferase reporter assay, and Pearson's χ2 test. The in vivo experiment was evaluated by establishing a subcutaneous tumorigenesis model. High-expressed circ_0000326 in BC tissues and cells was discovered, which was connected with an undesirable prognosis. Silencing of circ_0000326 visibly inhibited MCF-7 and BT549 cell growth, invasion, stemness, meanwhile declining the protein levels of SRY-related high-mobility group box gene 2 (SOX2) and octamer binding transcription factor 4 (OCT4). miR-9-3p was a sponger of circ_0000326, which was negatively regulated by circ_0000326. Moreover, YAP1 was confirmed as a target gene of miR-9-3p. circ_0000326 affected BC cell behaviors via mediating miR-9-3p and YAP1. Furthermore, circ_0000326 silencing prohibited tumor growth of BC in vivo. The research uncovered that circ_0000326 facilitated BC development via mediating the miR-9-3p/YAP1 axis.

Semaphorin 3A inhibits tumor progression via the downregulation of Lin28B in ovarian cancer

Semaphorin 3A (Sema3A) has recently been proven to play an essential role in tumorigenesis. Here, the role of Sema3A in ovarian cancer is explored. The prognostic value of Sema3A was evaluated using the Kaplan-Meier plotter database, and stable expression cells were established by the delivery of lentivirus harboring SEMA3A cDNA or shRNA into OVCA433 and SKOV3 cells, respectively. Then CCK-8 assay, colony-formation assay, wound-healing assay, and Transwell assay were utilized to verify the effect of Sema3A on tumorigenesis. Co-cultures of ovarian cancer cells (OVCA433 and SKOV3) with a conditional medium collected from the established cells were further utilized to confirm the function of Sema3A. Then, the RNA-seq assay was adopted to explore the underlying mechanism. The results demonstrated that low expression of Sema3A was predictive of poor overall survival in patients with ovarian cancer. Functional experiments revealed that Sema3A inhibited proliferation, migration, and invasion in ovarian cancer cells. Secreted Sema3A in a conditioned culture medium also exhibited an anti-tumor effect in ovarian cancer cells. RNA-seq assay suggested that focal adhesion and Lin28B were involved in regulating Sema3A. Rescue assays further verified that Lin28B/ROCK1 axis was vital in the regulation of Sema3A and Lin28B significantly upregulated ROCK1 through let-7g microRNA. The presented data indicate that Sema3A inhibits proliferation and metastasis via the downregulation of Lin28B/ROCK1 in ovarian cancer.

Tumor suppressor miR-424-5p abrogates ferroptosis in ovarian cancer through targeting ACSL4

Ovarian cancer is the most lethal gynecological cancer. In spite of recent advances, clinical outcomes remain poor, urgently needing novel therapeutic approaches. Growing evidence indicates that microRNAs play crucial roles in ovarian carcinogenesis and progression and that ferroptosis serves as a novel tumor suppressor. However, the molecular mechanisms of miRNA-mediated ferroptosis regulation in ovarian cancer are still largely unknown. In the present study, we show that miR-424-5p negatively regulates ferroptosis by directly targeting ACSL4 in ovarian cancer cells. Upregulation of miR-424-5p suppressed ACSL4 by directly binding to its 3'-UTR, which subsequently reduced erastin- and RSL3-induced ferroptosis. Meanwhile, knockdown of miR-424-5p increased the sensitivity of ovarian cancer cells to erastin and RSL3. Furthermore, ACSL4 was upregulated in ovarian cancer tissues, and high ACSL4 expression predicted worse prognosis and sensitized ovarian cancer cells to erastin- and RSL3-induced ferroptosis. Importantly, decreases in lipid peroxides and ferroptotic cell death mediated by miR-424-5p could be abrogated by ACSL4 overexpression. Taken together, our findings demonstrate that miR-424-5p regulates ferroptosis by targeting ACSL4 in ovarian cancer cells and suggest a potential therapeutic approach for ovarian cancer.

Overexpression of BMP9 promotes ovarian cancer progression via Notch1 signaling

Cell proliferation and migration play important parts in ovarian cancer progression. BMP9, as one of the members of the TGF-β superfamily and BMP family, plays a diverse and significant array of biological roles, including cell differentiation, proliferation, apoptosis, tumorigenesis, and metabolism. However, the role and mechanism of BMP9 in ovarian cancer progression remains uncertain. We found that the expression of BMP9 was increased in human ovarian cancer cell lines, which induced Notch1 intracellular domain (NICD1) accumulation. And we also found the expression abundance of BMP9 is low in ovarian cancer cells. Thus, we generated recombinant adenoviruses overexpressing BMP9 to perform the research. We found that overexpression of BMP9 promoted ovarian cancer cell proliferative viability, cell cycle progression, cell migration in vitro, and accelerated subcutaneous tumor growth in vivo, which was inhibited by dominant-negative mutant Notch1 recombinant adenoviruses. Besides, we also demonstrated that silencing of BMP9 by recombinant adenoviruses inhibited ovarian cancer cell viability and migration in vitro. Additionally, BMP9-induced ovarian cancer cell progression also involved the elevation of HES2, c-Myc, MMP9, and Cyclin D1, as well as repressed expression of p27. Together, these results revealed that BMP9 acts as a promoting factor in ovarian cancer progression, and overexpression of BMP9 promotes ovarian cancer progression and growth via Notch1 signaling. Thereby our research may provide new insight into the pathogenesis of ovarian cancer and BMP9-Notch1 signaling may serve as a novel therapeutic target axis for ovarian cancer treatment.

miR-485-5p improves the progression of ovarian cancer by targeting SRC in vitro and in vivo

miR-485-5p is involved in many tumors, but its role in ovarian cancer has been rarely reported. This paper mainly studied the expression and mechanism of miR-485-5p in ovarian cancer. The expression of miR-485-5p in ovarian cancer was compared with that in adjacent tissues, and the expression of miR-485-5p in various ovarian cancer cell lines was detected by RT-PCR. miR-485-5p mimics and pcDNA plasmid or pcDNA-SRC were transfected into SKOV3 cells. Cell proliferation was detected by CCK-8, cell cycle and apoptosis were detected by flow cytometry, and cell migration and invasion were detected by wound assay and transwell assay. Nude mice were inoculated with SKOV3 cells to detect the effect of miR-485-5p on xenograft. SRC and its downstream proteins were detected by western blot. Our data suggested that the expression of miR-485-5p was low in ovarian cancer tissues and ovarian cancer cell lines. miR-485-5p mimics can inhibit the proliferation, migration, invasion, and the induction of cycle arrest and apoptosis of SKOV3 cells in vivo and in vitro, and pcDNA-SRC can reverse the effect of miR-485-5p mimics on ovarian cancer. Our findings suggest that miR-485-5p could inhibit the progression of ovarian cancer by targeting SRC, which might be a new target for ovarian cancer therapy as the SRC inhibitor.

miR-193a-5p suppresses cell proliferation and induces cell apoptosis by regulating HOXA7 in human ovarian cancer

Ovarian cancer is one of the most common malignancies in women in the world. MicroRNAs (miRNAs) were identified as a group of regulators that played important roles in the progression of cancer development. The main purpose of this study was to investigate the functional mechanism of microRNA-193a-5p (miR-193a-5p) in human ovarian cancer. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the RNA levels of miR-193a-5p and homeobox genes A7 (HOXA7). Western blot assay was performed to determine the protein level of HOXA7. The interaction between miR-193a-5p and HOXA7 was predicted by online software starBase v3.0, and then verified by the dual luciferase reporter assay. The cell proliferation and apoptosis rate were examined by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) and colony formation assay as well as flow cytometry analysis. We found out that the expression level of miR-193a-5p was decreased in human ovarian cancer tissues and cells. The overexpression of miR-193a-5p inhibited cell proliferation and induced apoptosis in human ovarian cancer. Interestingly, miR-193a-5p reduced the expression of HOXA7 by binding to 3'-untranslated region (3'-UTR) of HOXA7 mRNA. As expected, the knockdown of HOXA7 also suppressed cell proliferation and promoted apoptosis in human ovarian cancer. Besides, the upregulation of HOXA7 reversed the effect of miR-193a-5p on human ovarian cell proliferation and apoptosis. Our findings confirmed that miR-193a-5p inhibited cell proliferation and induced apoptosis through the downregulation of HOXA7 in human ovarian cancer, providing a theoretical value for the therapy of human ovarian cancer.

LncRNA CDKN2B-AS1 promotes the progression of ovarian cancer by miR-143-3p/SMAD3 axis and predicts a poor prognosis

Long noncoding RNAs (LncRNAs) show great potential as the therapeutic targets attributing to their implication in the progression of various human cancers, including ovarian cancer (OC). Here, we aimed to explore the biological function of lncRNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) in OC and its mechanism of action. The abundances of CDKN2B-AS1, miR-143-3p, and SMAD3 mRNA were determined by quantitative real-time polymerase chain reaction (qRT-PCR). Cell Counting Kit-8 (CCK8) was performed to analyze cell proliferation. Cell apoptosis was assessed by flow cytometry and western blot analyses. Transwell assay was utilized to analyze cell migration and invasion abilities. Tumor xenograft was performed to confirm the role of CDKN2B-AS1 in ovarian tumor growth in vivo. The protein level of SMAD3 was examined by western blot assay. The interaction between CDKN2B-AS1 and miR-143-3p, or miR-143-3p and SMAD3 was demonstrated by bioinformatic, luciferase reporter, qRT-PCR and western blot analyses. CDKN2B-AS1 was upregulated in OC and correlated with clinicopathologic features. The knockdown of CDKN2B-AS1 hampered the development of OC, as reflected by the suppression of cell proliferation, migration, and invasion, and the enhancement of cell apoptosis, whereas the effects could be rescued by the overexpression of SMAD3. The absence of CDKN2B-AS1 blocked tumor growth in vivo. CDKN2B-AS1 served as a molecular sponge for miR-143-3p, leading to the derepression of miR-143-3p target SMAD3, which eventually triggered the progression of OC. In conclusion, CDKN2B-AS1 promoted tumor growth, invasion, and migration of OC by regulation of miR-143-3p/SMAD3 axis, hinting that CDKN2B-AS1 might be a potential biomarker for OC diagnosis and treatment.

Endostar blocks the metastasis, invasion and angiogenesis of ovarian cancer cells

Endostar (ES) inhibits metastasis in some tumors, but its role in ovarian cancer invasion has not been elucidated. In this study, the effects of ES on ovarian cancer cells were further analyzed, to excavate an effective strategy for treating ovarian cancer. Ovarian cancer cell lines (SKOV3 and HO-8910PM) were treated with different concentrations of ES. Cell activity and half-maximal inhibitory concentration (IC50) detected by MTT were used for subsequent experiments. The migration and invasion abilities of treated cells were detected by wound healing and Transwell assays. The expressions of epithelial-mesenchymal transition (EMT)-related proteins in treated cells were determined by western blot analysis. Moreover, in vitro angiogenesis, the expressions of related proteins in treated cells and STAT3, and PD-L1 expressions were determined. We found that with the increase of ES concentrations, the cell activity showed a decreasing trend, and that the compositive IC50 of SKOV3 and HO-8910PM was 50 μg/ml. Moreover, ES observably inhibited migration, invasion, and EMT of ovarian cancer cell lines. In angiogenesis experiments, the angiogenesis ability and the expressions of related proteins in ovarian cancer cell lines were downregulated after ES treatment. Furthermore, ES reduced the expression of PD-L1 and suppressed the phosphorylation of STAT3 in ovarian cancer cell lines. ES blocked the metastasis, invasion, and angiogenesis of ovarian cancer cells by suppressing the activation of PD-L1 and STAT3, which might be considered as the potential mechanism of ES in the treatment of ovarian cancer.

Quality of life and ability to work in ovarian cancer patients in Slovakia

The aim of this paper was to find out the association of relevant factors on health-related quality of life (HRQOL) among ovarian cancer patients and their ability to work. Analyzed data were prospectively collected on 123 ovarian cancer patients enrolled across multiple oncology practices in Slovakia. We examined knowledge about the disease, negative perceptions related to health care, ability to work and social and economic ranking. HRQOL measurements included quality of life based on a numeric scale (1-worst, 10-best) and selected aspects from QoL-Ov28 questionnaire. We have used non-parametric Friedman and Dunne pairwise comparison tests to detect differences in HRQOL and the ability to work. Spearman correlation was used to measure the strength of association between variables. With hindsight, patients identified first signs of disease 3.6 months prior to diagnosis, with median duration of disease being 3.1 years. HRQOL was significantly different at various points during cancer journey; between current state and at diagnosis (4.19), between current state and at time without cancer or at time in full health (8.94, 9.52 respectively). Similarly, significant differences were noted in patients' current work ability (WA) compared to WA at diagnosis, or at time without cancer or in full health (4.2, 9.07, 9.58). The highest correlation of HRQOL was found in relation to current ability to work (r = 0.87) and in impact of cancer treatment (r = 0.66). Medium correlation was noted with visits to oncology clinics, knowledge about cancer, salary, future expectations or perceived quality of life of relatives (r < 0.51). Low correlation (r < 0.3) was found with other aspects related to healthcare (nursing care, general practitioner appointments) or demographics (age, number of children) and others. Patients were willing to pay monthly for curative treatment €191.84 from an average monthly salary €470.84 (41%). Ovarian cancer diagnosis has a significant impact on HRQOL and WA and both are positively highly correlated. Ovarian cancer patients are willing to give significant share of their monthly salary for treatment leading to cure.

MicroRNA-506-3p inhibits proliferation and promotes apoptosis in ovarian cancer cell via targeting SIRT1/AKT/FOXO3a signaling pathway

Ovarian cancer (OC) is one of the most common tumors in females. Growing evidence shows that microRNA-506-3p (miR-506-3p) is downregulated in OC tissues. The purpose of this study was to investigate the mechanism of miR-506-3p in modulating OC. Quantitative reverse transcriptase PCR (qRT-PCR) was employed to investigate the expression of miR-506-3p and its target in OC tissues or cell lines. CCK-8 or colony formation assay was used to examine cell viability or proliferation, respectively. Flow cytometry was demonstrated to detect cell apoptosis. Western blot was then applied to analyze underlying mechanisms. The potential target of miR-506-3p was examined via luciferase reporter assay. MiR-506-3p was significantly downregulated in both human OC tissues and cell lines. Overexpression of miR-506-3p not only decreased cell viability of OC cell lines but also promoted cell apoptosis, thus inhibiting OC progression. Moreover, SIRT1 (Sirtuin 1) was found to be a direct target of miR-506-3p, and SIRT1 expression was negatively regulated by miR-506-3p in OC cell lines. Further investigation revealed that overexpression of SIRT1 could promote cell viability as well as inhibit cell apoptosis, showing the reversed effect on OC progression compared to miR-506-3p. Lastly, AKT (Protein kinase B) /FOXO3a (Forkhead box O3) signaling pathway was inactivated by miR-506-3p while activated by SIRT1, relating to regulation of miR-506-3p on OC progression. Our results revealed a novel mechanism by which miR-506-3p inhibited proliferation while promoted apoptosis of OC via inactivation of SIRT1/AKT/FOXO3a signaling pathway, suggesting that miR-506-3p might be a potential target for OC.

MiR-34a-5p/PD-L1 axis regulates cisplatin chemoresistance of ovarian cancer cells

Ovarian cancer is the most lethal gynecologic malignancy in women with an increasing number of cases worldwide. Chemoresistance is the main obstacle for ovarian cancer treatment during clinical therapy. Previous studies found that programmed cell death 1 ligand 1 (PD-L1) was associated with chemoresistance of cancer. However, there were little reports about the function of PD-L1 involved in chemoresistance of ovarian cancer. In our study, cisplatin (DDP)-resistant SKOV3 and A2780 ovarian cancer cell lines (SKOV3/DDP and A2780/DDP) were established. We found that the expression of PD-L1 was increased and miR-34a-5p was decreased in DDP-resistant cells. PD-L1 silencing inhibited chemoresistance of DDP-resistant ovarian cancer cells to DDP, as evidenced by decreased proliferation, G1-phase cell cycle arrest and increased apoptosis. Western blot assay showed that in the presence of DDP, PD-L1 silencing decreased multidrug resistance protein 1 and Cyclin D1 protein levels, whereas increased cleaved-caspase-3 and cleaved-PARP protein levels in these cells. Moreover, we demonstrated that miR-34a-5p negatively regulated the expression of PD-L1 by targeting its 3'-untranslated region. The effects of miR-34a-5p mimic on DDP-treated SKOV3/DDP cells were reversed by the overexpression of PD-L1. Moreover, the tumorigenicity of DDP-resistant ovarian cancer cells in nude mice treated with DDP was attenuated by miR-34a-5p in vivo. The combined data indicate that miR-34a-5p/PD-L1 axis regulates DDP chemoresistance of ovarian cancer cells, providing a deeper insight into the treatment for ovarian cancer.

High GNG13 expression is associated with poor survival in epithelial ovarian cancer and breast cancer

Recently, change in the GNG13 expression has been shown to result in multiple congenital malformations and sexual reversal, and it was also found in the brain. The aim of this study was to measure the expression levels in epithelial ovarian cancer (EOC) and breast cancer (BC) and assess their value as a potential prognostic marker. The correlation of GNG13 protein expression was detected by immunohistochemistry (IHC) in 119 EOC and 125 BC tissues. Assessment of the associations between GNG13 levels and various clinicopathological features was identified, the relationship between GNG13 and prognosis in BC and EOC patients was analyzed using online resources of Oncomine and Kaplan-Meier plotter. Protein expression levels of GNG13 were both significantly lower in BC and EOC compared with normal tissues (p<0.0001 and p<0.001, respectively). Among the clinicopathological characteristics of BC, tumor grade (p=0.001) and TNM stage (p=0.001) were significantly associated with low expression of GNG13. While in EOC, low expression of GNG13 was significantly related to FIGO stage (p=0.001), presence of metastasis (p=0.001), and CA125 (p=0.001). Our data suggest that GNG13 expression maybe as a new inhibitor, which can strongly inhibit metastasis and partially attenuates tumor growth in EOC and BC.

High-grade serous ovarian carcinoma and detection of inactivated BRCA genes from biopsy material of Slovak patients

Ovarian cancer is the leading cause of mortality among all gynecological cancers in developed countries and its most common and most lethal type is the high-grade serous ovarian carcinoma (HGSC). At the molecular level, nearly half of all HGSCs exhibit ineffective homologous DNA recombination and disruption of DNA damage/repair pathway inactivation caused often by BRCA1 and BRCA2 gene mutation. Recently, the detection of BRCA1/2 mutations became important for personalized treatment of HGSC patients with the PARP-inhibitors in the defined clinical setting of relapse after positive adjuvant platinum-based chemotherapeutic response. Based on the selection of patients by regional oncologists, we attempted to verify the possibilities of BRCA1/2 mutation testing on archival formalin-fixed paraffin-embedded (FFPE) biopsy material from regional hospitals. In the study we used: a/ FFPE tumor resections of 97 patients sent to our laboratory, originally stored in archives of regional departments for a period of 1-3 years and retrieved on the principle to contain a maximum of non-necrotic tumor tissue, b/ next-generation sequencing (NGS) assay covering all known mutations in the BRCA1/2 genes on MiSeq (Illumina® platform), and c/ Sophia DDM® bioinformatics platform. After processing of FFPE samples, 5 cases were excluded due to the insufficient genomic DNA quantity. Bioinformatics results of NGS analyses of 92 patients' samples indicated 17.39% pathogenic mutations and 32.61% potentially pathogenic mutations in genes BRCA1/2. Overall, 50% pathogenic and potentially pathogenic mutations were detected in the patient's cohort. The relatively high incidence of BRCA1/2 mutations in our series may be influenced by various indicators including the selection of patients based on adjuvant therapy response as well as regional or population heterogeneity in their frequency. Based on the interdisciplinary cooperation, the use of archival biopsy material processed primarily and stored for a longer period in different laboratories without uniformly defined pre-analytical conditions allows identifying the HGSC patients who might better respond to the PARP-inhibition therapy.

CCAAT enhancer binding protein α suppresses proliferation, metastasis, and epithelial-mesenchymal transition of ovarian cancer cells via suppressing the Wnt/β-catenin signaling

CCAAT enhancer-binding protein alpha (CEBPA, also known as C/EBPα) is a transcription factor that plays an essential role in regulating terminal differentiation and cell proliferation of many tissues. The objective of this study was to explore the potential function of CEBPA in ovarian cancer. The expression of CEBPA in ovarian cancer samples and adjacent normal tissues was evaluated by qRT-PCR. The putative role of CEBPA in ovarian cancer cells was evaluated by immunohistochemistry, western blot, cell viability assay, BrdU incorporation assay, soft agar colony formation assay, Transwell cell migration and invasion assay, tumor xenograft formation, and lung metastasis model. We found that CEBPA was downregulated in ovarian cancer samples and predicted a poor prognosis. CRISPR/Cas9-mediated CEBPA knockout promoted proliferation, anchorage-independent growth, migration, invasion, and EMT of ovarian cancer cells, while enforced CEPBA expression suppressed proliferation, anchorage-independent growth, migration, invasion, EMT, tumor xenograft growth, and lung metastasis of ovarian cancer cells. Furthermore, we found that the knockout of CEBPA activated Wnt/β-catenin signaling in ovarian cancer cells, while CEBPA overexpression suppressed Wnt/β-catenin activation. Our data indicated that CEBPA acted as a tumor suppressor in ovarian cancer, and might be a potential prognostic marker for ovarian cancer treatment.

The results of multigene panel sequencing in Slovak HBOC families

Hereditary breast and ovarian cancer (HBOC) is primarily associated with mutations in the BRCA1/2 genes. However, causal variants in other high, moderate, and low penetrance genes proportionally increase the risk of breast/ovarian cancer. This study aims to provide data about the mutation spectrum of HBOC-associated genes in Slovak HBOC families and estimate the ratio of BRCA versus non-BRCA causal variants. We used panel sequencing containing 22 high/moderate-risk susceptibility genes and parallel MLPA analysis of BRCA1/2, CHEK2 genes, to analyze 94 individuals with a strong family/personal history of breast and/or ovarian cancer. The analyzed group consisted of 80 patients diagnosed with cancer (85.1%) and 14 healthy individuals (14.9%) with a positive family history of HBOC syndrome. In total, we have identified 22 causal DNA variants (23.4%) showing 15 primary findings in BRCA1/2 genes (68.2%) and 7 positive secondary findings in CHEK2, PALB2, CDH1, and MUTYH genes (31.8%). The most frequent pathogenic alterations were BRCA1 mutations c.181T>G and CNV variant (c.5573-?_c.5701+?)del, known as deletion of exons 21-22. Besides known mutations, the BRCA1 variant c.2794del (p.Val932Leufs*68) and variant c.2480dup (p.Tyr827*) in the CDH1 gene represent the novel, previously unpublished variants that might be population-specific. In conclusion, we provide the first report of multigene panel testing in Slovak HBOC families demonstrating that almost one-third of pathogenic mutations are situated in susceptibility genes other than BRCA1/2. Although multigene panel testing requires precise data filtration and interpretation, it might bring the relevant data for clinical management of the patients.

LncRNA HOTAIR regulates anoikis-resistance capacity and spheroid formation of ovarian cancer cells by recruiting EZH2 and influencing H3K27 methylation

This study aims to investigate the role of the long non-coding RNA (lncRNA) HOX transcript antisense RNA (HOTAIR) in the regulation of anoikis resistance of ovarian cancer cells, a prerequisite for metastasis and chemoresistance in ovarian cancer cells. Ovarian cancer SKOV3 cells were cultured in an ultra-low attachment system to establish an anoikis model. The relationship between cellular anoikis capability and HOTAIR expression level was studied by flow cytometry and RT-PCR. The ability of spheroid formation, migration, and invasion of the suspended cells was assessed following the knockdown of HOTAIR expression. The expression of EZH2, H3K27me3, representative targets of EZH2, and anoikis-related biomarkers was also detected. An increase in the duration of suspension culture time rendered the SKOV3 cells anoikis-resistant with a significantly lower apoptotic rate compared to the adherent cells. HOTAIR expression in the suspension cells increased significantly, while that in the adherent cells did not. Following small interfering RNA (siRNA)-mediated knockdown of HOTAIR expression, the abilities of anoikis resistance, migration, and invasion decreased in the suspension cells. Knockdown of HOTAIR levels also reduced the spheroid forming ability of the tumor cells in continuous suspension cultures. Moreover, EZH2 expression correlated with HOTAIR expression, thus regulating the expression of miR-193a and DOK2 via introducing H3K27me3. Western blot analysis of anoikis-related markers showed that N-cadherin, ZEB1, and TWIST1 were downregulated following inhibition of HOTAIR, while E-cadherin and ErbB3 were upregulated. In conclusion, HOTAIR enhances the anoikis resistance and spheroid forming ability of ovarian cancer cells by recruiting EZH2 and influencing H3K27 methylation that may contribute to migration, invasion, and chemoresistance of ovarian cancer cells.

Outcomes of icotinib combined with concurrent chemoradiotherapy in locally advanced cervical cancer

Concurrent chemoradiotherapy (CRT) based on cisplatin is recognized as the current standard treatment for locally advanced cervical cancer. The treatment of cervical cancer has reached a plateau in the last 20 years. Previous studies have proven that the epidermal growth factor receptor is correlated with chemo- and radioresistance and treatment failure. Hence, the purpose of this study was to investigate the efficacy and safety of icotinib combined with CRT in the treatment of locally advanced cervical cancer. Eligibility criteria included patients treated in the radiotherapy department of Taizhou Central Hospital of Zhejiang Province for stage IIB to IIIB cervical cancers who had not received anti-tumor treatment before and a performance status of 0 to 2. Patients were given icotinib 125 mg three times a day for 6 weeks, which was one week before the start of radiotherapy (500 centigrays in 28 fractions) and chemotherapy (40 mg/m2 administered weekly for 3-5 cycles). There were 29 patients who completed the I+CRT treatment, and it was tolerated well. The median follow-up time was 50 months and 27 patients (93.10%) achieved complete responses. The 5-year cumulative overall survival rate and disease-free survival rate were 58.4% and 60.9%, respectively. The treatment with I+CRT is safe and effective for locally advanced cervical cancer. As far as we know, this is the first study to report the 5-year survival rate of locally advanced cervical cancer with targeted therapy combined with chemoradiotherapy.

LncRNA CBR3-AS1 predicts a poor prognosis and promotes cervical cancer progression through the miR-3163/LASP1 pathway

LncRNA carbonyl reductase antisense RNA 1 (CBR3-AS1) is increased in cervical cancer and predicts poor prognosis. This study aims to investigate the underlying mechanism of lncRNA CBR3-AS1 in cervical cancer. LncRNA CBR3-AS1 and LASP1 expressions were significantly elevated in cervical cancer tissue and cells, whereas miR-3163 expression was significantly decreased in cervical cancer tissue and cells. High lncRNA CBR3-AS1 expression and LASP1 expression showed a lower overall survival rate, whereas high miR-3163 expression showed a higher overall survival rate. Correlation between clinicopathological parameters of cervical cancer patients and lncRNA CBR3-AS1, miR-3163, LASP1 expressions indicated that the expressions of lncRNA CBR3-AS1, miR-3163, and LASP1 were closely related with distant metastasis and lymphatic metastasis of cervical cancer. LncRNA CBR3-AS1 knockdown suppressed cervical cancer cell viability and inhibited cancer stem cell-like properties. Besides, we identified that lncRNA CBR3-AS1 interacted with miR-3163, and miR-3163 targeted to LASP1. Moreover, the correlation between lncRNA CBR3-AS1 and miR-3163, as well as the correlation between miR-3163 and LASP1 was confirmed. Finally, lncRNA CBR3-AS1 knockdown inhibited tumor growth and suppressed cancer stem cell-like properties of cervical cancer in vivo. Taken together, high expression of lncRNA CBR3-AS1 predicts poor prognosis in cervical cancer, and the lncRNA CBR3-AS1/miR-3163/LASP1 pathway plays a vital function in the modulation of cervical cancer cell proliferation and cancer stem cell-like properties.

EFEMP2 increases the invasion ability of cervical cancer cells by promoting EMT via the Raf/MEK/ERK signaling pathway

EFEMP2 has been reported as a candidate oncogene. To investigate the role of EFEMP2 in cervical cancer cell proliferation and invasion, the mRNA and protein expressions of EFEMP2 in 5 different cervical cancer cell lines were detected. And then the effects of up- or down-regulation of EFEMP2 expression on the biological behavior of cervical cancer cells were further investigated by transfection experiments and cell function assays in vitro and in vivo. The results revealed that EFEMP2 was highly expressed in highly invasive Ca Ski cells and lowly expressed in less invasive HT-3 cells. When EFEMP2 was knocked down, the proliferation and invasion ability of cervical cancer cells were also reduced, accompanied by the decreased expression of MMP-1, MMP-13, MMP-3, and MMP-10, meanwhile, the EMT process was blocked and the Raf/MEK/ERK signaling pathway was inhibited. On the contrary, the upregulation of EFEMP2 could promote the proliferation and invasion of cervical cancer cells by inducing EMT and activating the Raf/MEK/ERK pathway. In conclusion, EFEMP2 could increase the invasion ability of cervical cancer cells by upregulating the expression of MMP-1, MMP-13, MMP-3, and MMP-10 and inducing the EMT process through the Raf/MEK/ERK pathway. EFEMP2 played a promoting role in the development of cervical cancer and provided a potential therapeutic target for inhibiting the invasion and metastasis of cancer cells and improving the prognosis of cervical cancer patients.

Vitamin D supplementation in cancer prevention and the management of cancer therapy

EGLN2 attenuates ovarian cancer malignancy via ferroptosis activation

Growing evidence indicates that ferroptosis is pivotal in the development and progression of ovarian cancer (OC). However, the function of EGLN2 in OC remains unclear. In this study, we observed that EGLN2 is expressed at low levels in OC tissues and is associated with a favorable prognosis in early-stage patients based on data from TCGA and GTEx public databases. Compared with those in normal ovarian epithelial cell lines, EGLN2 mRNA and protein levels were significantly lower in OC cell lines. In vitro functional experiments revealed that EGLN2 overexpression reduced proliferation, increased the intracellular levels of iron ions, reactive oxygen species, lipid peroxidation, and mitochondrial oxidative phosphorylation, and inhibited the Warburg effect. Mechanistically, EGLN2 inhibited the expression of HIF-1α, which binds to the promoter of ceruloplasmin (CP), reducing the proliferation of and inducing ferroptosis in OC cells. A subcutaneous xenograft model in nude mice demonstrated that EGLN2 overexpression inhibited HIF-1α, promoted ferroptosis, and inhibited OC cell growth. In summary, EGLN2 suppressed CP transcription and increased ferroptosis in OC cells. These findings provide new insights into OC development and open avenues for innovative therapies.

miR-124 delivered by BM-MSCs-derived exosomes targets MCT1 of tumor-infiltrating Treg cells and improves ovarian cancer immunotherapy

Metabolic rewiring of tumor cells leads to an enrichment of lactate in the tumor microenvironment (TME). This lactate-rich environment of solid tumors has been reported to support tumor-infiltrating regulatory T (Treg) cells. Therefore, agents that modify the lactate metabolism of Treg cells have therapeutic potential. Monocarboxylate transporter 1 (MCT1), which Treg cells predominantly express, plays an essential role in the metabolism of tumor-infiltrating Treg cells. In this study, we show that miR-124 directly targets MCT1 and reduces lactate uptake, eventually impairing the immune-suppressive capacity of Treg cells. Particularly, exosomal miR-124 derived from bone marrow mesenchymal stromal cells (BM-MSCs) slows tumor growth and increases response to PD-1 blockade therapy. These data indicate a potential treatment strategy for improving immune checkpoint blockade therapy using miR-124-carried BM-MSCs-derived exosomes.

CCL18 derived from M2‐polarized tumor‐associated macrophages promotes endometrial cancer progression by activating the TWIST1/HMGA1 axis

CCL18, originating from M2-polarized tumor-associated macrophages (M2-TAMs) is recognized for its vital role in endometrial cancer (EC) development. Nonetheless, its precise mechanisms remain largely undefined. The primary objective of this research was to elucidate the underlying mechanism of M2-TAM-isolated CCL18 in EC progression. TWIST1 and HMGA1 expressions were assessed in EC tissues and cells by qRT-PCR or western blotting. M2 macrophages were differentiated from human monocyte THP-1 cells, and characterized via flow cytometry and western blotting. CCL18 levels were evaluated using western blotting and ELISA assay. CCK8, Transwell, and wound healing assays were employed to assess EC cell vitality, invasion, and migration, respectively, while western blotting was utilized to measure related protein markers. The binding relationship between TWIST1 and HMGA1 was validated via ChIP and dual-luciferase reporter assays. TWIST1 and HMGA1 were increased in EC tissues and cells. After being treated with M2-TAM-isolated CCL18, EC cell vitality, migration, and invasion were enhanced. Additionally, CCL18 derived from M2-TAM upregulated TWIST1 levels in EC cells. Further mechanistic analyses unveiled that TWIST could positively regulate HMGA1 in EC cells. Notably, HMGA1 knockdown restrained the malignancy of EC cells, which was reversed by TWIST1 overexpression. M2-TAM-isolated CCL18 facilitated EC progression by activating the TWIST1/HMGA1 axis. These observations might offer new directions for developing targeted curative interventions for EC.

TRIM44 facilitates ovarian cancer proliferation, migration, and invasion by inhibiting FRK

Ovarian cancer (OC) is the leading cause of gynecologic cancer-related death in the world. Accumulating evidence indicated the important role of TRIM44 in cancer development. However, how TRIM44 displays in OC and the underlying mechanism remained unclear. TRIM44 and FRK expression in OC tissues and cell lines were investigated by western blot and RT-qPCR. Histotype of tissue samples and patients' data were analyzed. Kaplan-Meier Curve was performed to validate the effect of TRIM44. Colony formation assay, MTT assay, Transwell assay, and wound-healing assay were applied to elucidate the function of TRIM44 in OC cells. CHIP assay was used to explore the association between TRIM44 and FRK. Finally, we performed SKOV3 xenografts in Balb/c nude mice to further confirm the involvement of TRIM44 in OC development. We found TRIM44 highly expressed while FRK displayed low expression in OC cell lines and tissues. Moreover, analysis of histotype of tissues and patients' data and Kaplan-Meier Curve implied the important role of TRIM44 and FRK in tumor progression. Further in vitro study suggested that knocking down TRIM44 inhibited OC cells proliferation, migration, and invasion. Besides, FRK was identified as the target gene of TRIM44 in OC, and TRIM44 promoted OC cells proliferation, migration, and invasion by inhibiting FRK. Finally, in vivo animal experiment further confirmed the promotive effect of TRIM44 on OC progression. Our findings demonstrated that TRIM44 facilitated OC proliferation, migration, and invasion by inhibiting FRK, providing new insights for theoretical research and therapy of OC.

The impact of neo/adjuvant treatment choices on prognosis for surgically treated small-cell neuroendocrine carcinoma of the cervix

Small-cell neuroendocrine carcinoma of the cervix (SCNCC) is a rare and aggressive tumor with a poor prognosis. Surgical resection followed by adjuvant therapy is the standard treatment for early-stage disease but the influence of different neo/adjuvant treatment approaches remains unclear. Retrospectively, we collected patients' characteristics and treatments in two medical centers. Disease status and survival outcomes were renewed through follow-up. Statistics analysis mainly included Kaplan-Meier methods for survival curve estimation, log-rank test for survival curve comparison, and Cox proportional hazards models for independent prognostic factors prediction. Finally, 51 patients treated by radical surgery between January 2010 and April 2020 were enrolled with a median age of 50 years (range: 32-68). 12 (23.5%) patients were at stage IIIC1 according to the International Federation of Gynecology and Obstetrics (FIGO) 2018 staging systems and the rest were at the early stage. The mean tumor size was 3.6±1.3 cm. Pathological examination found 24 cases with pure SCNCC and 27 cases with admixed SCCC. 29 (56.9%) patients had deep stromal infiltration and 19 (37.3%) patients had lymphovascular space invasion. 34 (66.7%) patients received neo/adjuvant chemotherapy and pelvic radiation was conducted in 41 (80.39%) patients with a median dose of 46 Gy (range: 40-50.4 Gy). The median follow-up time was 25.0 months. The median disease-free survival (DFS) time was 23.0 months. 27 (52.9%) patients developed distant metastasis and 14 (27.5%) experienced local failure. The median overall survival (OS) was 32.0 months. Univariate and multivariate analysis showed neoadjuvant chemotherapy as negative (HR=2.081, 95% CI 1.030-4.203, p=0.041) and adjuvant chemotherapy (HR=0.409, 95% CI 0.213-0.784, p=0.020) as positive independent prognostic factor for DFS. For OS, only lymph node metastasis was confirmed as an independent prognostic factor in both univariate analysis (HR=1.528, 95% CI 1.011-2.308, p=0.044) and multivariate analysis (HR=1.697, 95% CI 1.041-2.768, p=0.034). In conclusion, for surgically treated SCNCC, adjuvant chemotherapy showed a positive influence on DFS while neoadjuvant chemotherapy harmed DFS. OS was unaffected by either treatment choice.

Resveratrol reduces lactate production and modifies the ovarian cancer immune microenvironment

Tumor cells show deregulated metabolism leading to an enrichment of lactate in the tumor microenvironment (TME). This lactate-rich environment has been reported to impair effector T cells. However, T-regulatory cells (Tregs) show metabolic advantages in lactate-rich TME that maintain a strong suppression of effector T cells, which leads to tumor immune evasion. Therefore, the glycolytic process of tumors could represent a therapeutic target, and agents that modify the energy metabolism of tumor cells have therapeutic potential. Resveratrol is a naturally occurring polyphenol that has been confirmed to suppress tumor cells' glycolytic metabolism. In this study, we show that resveratrol induces metabolic reprogramming in ovarian cancer cells. Resveratrol increases oxidative and decreases glycolysis, in association with decreased lactate production both in vitro and in vivo. Lactate reduction in TME weakens the suppressive function of Tregs, and subsequently restores anti-tumor immunity. Significantly, combined resveratrol and PD-1 blockade promote anti-tumor efficacy. These data suggest that resveratrol's anti-tumor actions in ovarian cancer could be explained, in part, through modification of the anti-tumor immunity, and indicate a novel treatment strategy for improving immune checkpoint blockade therapy using resveratrol.

The potential of ciRS-7 for predicting onset and prognosis of cervical cancer

Considering the potency of circRNAs in manifesting neoplastic progression, we attempted to explore the feasibility of applying ciRS-7 for diagnosis and prognosis estimation of cervical cancer (CC). Here 352 CC patients, 204 cervical intraepithelial neoplasia (CIN) patients and 227 healthy controls were recruited. The Kaplan-Meier survival curves were fitted to estimate associations of ciRS-7 expression with CC prognosis, and we also adopted receiver operating characteristic (ROC) curves to assess the diagnostic performance of ciRS-7 for CC. Meanwhile, endometrial stromal cell line (ESC) and 4 human CC cell lines (i.e. Hela, CaSki, C33A and SiHa) were also gathered. After transfection of pcDNA3.1-ciRS-7, impacts of overexpressed ciRS-7 on proliferation, apoptosis, invasion and migration of CC cells were assessed through performing colony formation assay, flow cytometry, transwell assay and wound healing assay. The results demonstrated that CC patients that highly expressed ciRS-7 were associated with high odds of large tumor size, advanced FIGO stage, deep invasion, metastatic lymph nodes and HPV infection (p<0.05). Furthermore, ciRS-7 excelled in differentiating CC patients from healthy controls and CIN patients than CEA and CA125 (p<0.05), and ciRS-7 combined with CA125 and CEA generated an optimum efficacy in diagnosing CC patients and CIN patients from healthy controls (p<0.05). Concerning in vitro experiments, elevating ciRS-7 expression significantly intensified proliferation and epithelial-mesenchymal transition (EMT) of CC cells (p<0.05), and also markedly suppressed apoptosis of the cells (p<0.05). In conclusion, ciRS-7 possessed great potential in clinical diagnosis of CC, given its involvement in modulating the activity of CC cells.

MCM2 promotes the stemness of endometrial cancer cells via the Akt/β-catenin pathway

Minichromosome maintenance complex component 2 (MCM2) is a member of the MCM family and is involved in various cancers. However, the role of MCM2 in endometrial cancer (EC) remains unclear. In this study, we aim to determine the biological function of MCM2 in EC cells and identify the potential underlying mechanisms. MCM2 expression and prognostic significance were analyzed in TCGA-UCEC datasets. Combining bioinformatics analyses and experiments, stemness-related molecules and phenotypes were examined to evaluate the impact of MCM2 on stemness in EC cells. The major findings of these analyses are as follows: 1) MCM2 is expressed at higher levels in EC tissues than in normal endometrial tissues. High expression of MCM2 is related to the characteristics of poorly differentiated EC. High MCM2 expression is correlated with poor overall survival in EC patients; 2) MCM2 knockdown was found to decrease sphere formation ability, downregulate the expression of stemness-related molecules, and reduce the proportion of CD133+ cells, while MCM2 overexpression elicited the opposite effect in EC cells; 3) MCM2-mediated stemness features are dependent on the activation of Akt/β-catenin signaling pathways; and 4) MCM2 knockdown increases cisplatin sensitivity in EC cells. MCM2 regulates stemness by regulating the Akt/β-catenin signaling pathway in EC cells.

miRNA-329-3p suppresses proliferation and metastasis of endometrial carcinoma through downregulating E2F1

Existing evidences have revealed the crucial roles of E2 promoter binding factor-1 (E2F1) during the tumorigenesis and progression process of multiple human tumors. However, the expression patterns, biological functions, as well as the underlying molecular mechanism of E2F1 in endometrial carcinoma yet remain largely unclear. The expression patterns and clinical prognostic value of E2F1 in endometrial carcinoma were evaluated using bioinformatics methods. Protein and mRNA, miRNA expression levels in tissues and cells were measured using immunohistochemistry, western blotting, and qRT-PCR assays. Cell viability and cell cycle distribution were examined using CCK-8 assay and flow cytometry, respectively. Scratch healing assay and Transwell assay were applied to measure cell migration and invasion ability. Bioinformatic analysis and luciferase reporter assays were conducted to confirm the targeting relationship between E2F1 and miR-329-3p. Moreover, a series of in vitro and in vivo functional experiments were employed to evaluate the effect of the miR-329-3p/E2F1 axis on cell growth and metastasis. Clinically, E2F1 was aberrantly expressed in endometrial carcinoma tissues and was correlated with advanced FIGO stage, histological type, p53 mutation, poor survival, and degree of tumor cell differentiation. ROC curves analysis also reveals that E2F1 has a high AUC value (up to 0.952, 95% CI: 0.915-0.988), indicating the promising diagnostic value of E2F1 level in endometrial carcinoma. In addition, in vitro gain and loss-of-functional experiments verified that high E2F1 can promote cell proliferation, cell cycle, migration, invasion, and EMT process. In-depth mechanism studies revealed that E2F1 was a downstream target gene of miR-329-3p, and miR-329-3p overexpression could effectively abrogate its promotion of cell malignant biological behavior. Collectively, our findings suggested that the miR-329-3p/E2F1 axis plays a crucial role in the progression of endometrial carcinoma, indicating that E2F1 can be considered a promising diagnostic and prognostic biomarker for endometrial carcinoma patients.

Optimization of endometrial cancer organoids establishment by cancer-associated fibroblasts

Most endometrial cancers (EC) are diagnosed at an early stage with a favorable prognosis. However, for patients with advanced or recurrent disease, the chemotherapy response rate and overall survival remain poor. A novel in vitro model, tumor organoids, has important value in providing a more individualized treatment plan for tumor patients. However, the slow growth of the established EC organoid seriously hinders the application of EC organoids. Cancer-associated fibroblasts (CAFs), the main component of tumor stroma, have been reported to promote the proliferation of endometrial cancer cell lines and primary endometrial cancer cells in vivo and in vitro. Therefore, we optimized the current endometrial cancer organoid by introducing CAFs isolated from EC lesions. Here we developed long-term expandable organoids from endometrial cancer lesions, which show disease-associated traits and cancer-linked mutations. Based on the co-culture of CAFs and endometrial cancer organoids, we found that CAFs could promote the growth of endometrial cancer organoids, might by secreting factors according to the result that CAFs could also promote the growth. Our research provided a more promising model for the basic and preclinical study of endometrial cancer.

A metabolomic signature of FIGO stage I and II endometrial cancer

Endometrial cancer (EC) is a malignant tumor of the female reproductive tract. Due to its rapid growth and invasiveness, EC is currently the only gynecological neoplasm with rising incidence and mortality rates. It is of great significance to explore the metabolomics signature of stage I and II EC for the diagnosis and treatment. A mass spectrometry-based untargeted metabolomics approach was used to explore preoperative serum metabolites in the normal and stage I and II EC patients. The metabolites were mapped to the Ingenuity pathway analysis (IPA) database to determine the potential biomarkers and metabolic pathways that differ between EC patients and healthy controls. The top analysis-ready molecules of upregulated D-glucose thiamine and downregulated cholesterol, arachidonic acid, palmitic acid, oleic acid, stearic acid, linoleic acid may be the most related metabolites. These potential biomarkers have essential functions in regulating vital metabolic pathways associated with stage I and II EC. Additionally, our pathway analysis revealed five significantly related pathways according to the metabolite differentials. Finally, the disease and function prediction of the initial pathway analysis suggested that small molecule biochemistry, lipid metabolism, and organismal injury and abnormalities were associated with EC cases. Over 25 metabolites were differentially expressed in stage I and II EC. In addition, the six most significant metabolites were related to stage I and stage II EC cases. Ingenuity pathway analysis revealed potential biomarkers and metabolic pathways revolved to EC. In this paper, candidate endogenous biomarkers were defined as the basis for disease diagnosis and individualized treatment monitoring and revealed the mechanism of EC occurrence and development.