Yan Zhang

p53 inhibits OTUD5 transcription to promote GPX4 degradation and induce ferroptosis in gastric cancer

AbstractBackgroundGastric cancer is one of the most prevalent malignant tumors within the digestive system, and ferroptosis playing a crucial role in its progression. Glutathione peroxidase 4 (GPX4), a key negative regulator of ferroptosis, is highly expressed in gastric cancer and contributes to tumor growth. Targeting the regulation of GPX4 has emerged as a promising approach to induce ferroptosis and develop effective therapy for gastric cancer.MethodsTo confirm that OTUD5 is a deubiquitinase of GPX4 and regulates ferroptosis, we performed Western blotting, Co‐IP, immunofluorescence, quantitative real‐time PCR, Ub assay and flow cytometry experiments. To explore the physiological function of OUTD5, we knocked out the Otud5 gene in the mouse gastric cancer cell line (MFC) using CRISPR‐Cas9 and eatablished the subcutaneous tumour model. Immunohistochemistry (IHC) analysis was used to inveatigate the pathological correlation in human gastric cancer.ResultsWe report that ovarian tumor domain‐containing 5 (OTUD5) interacts with, deubiquitylates and stabilizes GPX4. OTUD5 depletion destabilizes GPX4, promotes lipid peroxidation and sensitizes gastric cancer cells to ferroptosis. Moreover, the p53 activator nutlin‐3a suppresses OTUD5 transcription, leading to GPX4 degradation and ferroptosis of gastric cancer cells. Notably, only wild‐type p53 has the capacity to inhibit OTUD5 transcription, while p53 mutations or deficiencies correlate with increased OTUD5 expression, promoting gastric cancer progression. Additionally, OTUD5 silencing and nutlin‐3a‐induced GPX4 degradation enhances the sensitivity of gastric cancer cells to ferroptosis in vivo. Subsequently, the p53/OTUD5/GPX4 axis is confirmed in clinical gastric cancer samples.ConclusionCollectively, these findings elucidate a mechanism whereby p53 inactivation upregulates OTUD5 transcription to deubiquitylate and stablize GPX4, resulting in ferroptosis inhibition and gastric cancer progression. This discovery highlights the potential therapeutic value of targeting OTUD5 to promote ferroptosis in p53‐inactivated gastric cancer.Key points OTUD5 mediates GPX4 deubiquitination to regulate its stability. Deletion of OTUD5 promotes ferroptosis and inhibits tumor growth. Wild type p53 inhibits OTUD5 transcription, thereby promoting GPX4 degradation and inhibiting the development of gastric cancer. OTUD5, GPX4 expression and p53 activity are highly correlated and correlates with clinical progression in STAD.

Clinicopathological and prognostic significance of the microcystic elongated and fragmented pattern in endometrial cancer: a systematic review and meta-analysis

Objective The presence of the microcystic elongated and fragmented (MELF) pattern, distinguished by its microcystic, elongated and fragmented attributes, constitutes a common manifestation of myometrial invasion (MI) within endometrial carcinoma. However, the prognostic significance of this pattern has not been definitively established. Consequently, this research aimed to clarify the prognostic implications of the MELF pattern for individuals diagnosed with endometrial carcinoma. Design Systematic review and meta-analysis of observational clinical studies. Data sources An extensive review of the literature was conducted using reputable databases such as PubMed, Embase, Web of Science and the Cochrane Library, covering the period from January 2003 to October 2023. Search terms encompassed endometrial cancer and the MELF pattern. Eligibility criteria for selecting studies The inclusion criteria were patients who had undergone hysterectomy and whose pathology confirmed endometrial endometrioid carcinoma, with or without MELF infiltration. Data extraction and synthesis Two reviewers performed data extraction separately. The quality of the included studies was assessed using the Newcastle-Ottawa Scale (NOS). Stata V.17.0 software was used for statistical analysis. Results The meta-analysis incorporated 16 retrospective cohort studies. Employing a fixed-effects approach, the analysis demonstrated an association of the MELF pattern with reduced overall survival (HR 2.21, 95% CI 1.50–3.25, p=0.000) and lower disease-free survival rates among patients with endometrial cancer (HR 1.72, 95% CI 1.17 to 2.55, p=0.006). Furthermore, aggregated data revealed a linkage between the MELF pattern and significant MI, nodal metastasis, involvement of the lymphovascular space, penetration of the cervical stroma and progression to advanced stages of endometrial carcinoma. Conclusion The MELF pattern serves as a significant adverse prognostic factor in endometrial cancer, warranting increased attention.

Aberrant expression of UBE2C in endometrial cancer and its correlation to epithelial mesenchymal transition

Ubiquitin-conjugating enzyme E2C (UBE2C), its overexpression promotes tumor progression, is a key component of the ubiquitin conjugating proteasome complex. Epithelial-mesenchymal transition, which is lost epithelial features and gained mesenchymal features in some epithelial cancers, is involved in epithelial cancers’ invasiveness and metastasis. The aim of this study is to detect the expression of UBE2C, WNT5α, and E-cad in endometrial cancer (EC) and their clinical significance. The expression of UBE2C, WNT5α, and ZEB1 in 125 cases EC tissues were detected by immunohistochemistry. Patients clinicopathological, demography, and follow-up data were also collected. Positive rates of expression of UBE2C and ZEB1 were significantly higher in EC tissues when compared with the control tissues. The positive expression of UBE2C and ZEB1 were positively associated with tumor stages, local lymph node metastasis, and International Federation of Gynecology and Obstetrics (FIGO) stages. The positive rate of expression of WNT5a was significantly lower in EC tissues when compared with the control tissues. And positive expression of E-cad was inversely related to tumor stages, lymph node metastasis stages, and FIGO stages. Kaplan–Meier analyses demonstrated that positive expression of UBE2C or ZEB1 for EC patients had unfavorably overall survival time when compared with patients with negative expression of UBE2C or ZEB1. And EC patients with positive expression of WNT5a had favorably overall survival time when compared with EC patients with negative expression of WNT5a. Multivariate analysis demonstrated that positive expression UBE2C, WNT5α, and ZEB1, as well as FIGO stages were independent prognostic factors for EC patients. UBE2C, ZEB1, and WNT5a should be considered promising biomarkers for EC patients’ prognosis.

Molecular classification and adjuvant treatment in endometrioid endometrial cancer with microcystic elongated and fragmented (MELF) invasion pattern

To assess the characteristics, molecular classification, and role of adjuvant treatment in patients with endometrioid endometrial cancer (EEC) and microcystic elongated and fragmented (MELF) myometrial invasion pattern. This study included patients who were diagnosed with EEC with a MELF invasion pattern and underwent surgery from January 2019 to December 2023. We analyzed molecular classification, clinicopathological characteristics, and prognostic outcomes, including recurrence and adjuvant therapy. Out of 529 patients, 84 (15.9%) exhibited the MELF pattern, with 1 (1.2%) classified as POLE-mutation, 19 (22.6%) as mismatch repair deficient, 53 (63.1%) as no specific molecular profile, and 11 (13.1%) as p53-mutant subtype. Fifty (59.5%) patients with MELF invasion pattern received adjuvant chemotherapy (CT) ± external beam radiation therapy (EBRT), 19 (22.6%) received EBRT only, and 15 (17.9%) received no adjuvant treatment. Receiving adjuvant therapy was significantly associated with the risk level defined by the ESMO guideline for endometrial cancer (p = 0.002). With a median follow-up of 26 months (range: 1-59), the progression-free survival at 3-years for the MELF invasion patients was 91%. Seven patients with the MELF pattern experienced recurrence, of whom one was in stage IA (low risk, local recurrence) and did not receive any additional treatment, two were in stage IB (intermediate / high-intermediate risk, distant recurrence) and received EBRT only and the remaining four were in stage III to IV and had distant recurrence despite receiving adjuvant chemotherapy with or without EBRT. Among 43 intermediate- and high-intermediate-risk EEC patients, receiving CT ± EBRT was significantly associated with better DFS than without CT (p = 0.047). The presence of the MELF pattern in EEC should be incorporated into decision-making regarding adjuvant therapy. The use of adjuvant treatment should be tailored based on histology and molecular type.

A first-in-class POLRMT specific inhibitor IMT1 suppresses endometrial carcinoma cell growth

AbstractExploring novel molecularly-targeted therapies for endometrial carcinoma is important. The current study explored the potential anti-endometrial carcinoma activity by a first-in-class POLRMT (RNA polymerase mitochondrial) inhibitor IMT1. In patient-derived primary human endometrial carcinoma cells and established lines, treatment with IMT1 potently inhibited cell viability, proliferation, cell-cycle progression and motility, while inducing robust caspase-apoptosis activation. Treatment with the PLORMT inhibitor impaired mitochondrial functions, leading to mtDNA (mitochondrial DNA) transcription inhibition, mitochondrial membrane potential decline, reactive oxygen species formation, oxidative stress and ATP loss in the endometrial carcinoma cells. Similarly, POLRMT depletion, through shRNA-induced silencing or CRISPR/Cas9-caused knockout (KO), inhibited primary endometrial carcinoma cell proliferation and motility, and induced mitochondrial dysfunction and apoptosis. Importantly, IMT1 failed to induce further cytotoxicity in POLRMT-KO endometrial carcinoma cells. Contrarily, ectopic overexpression of POLRMT further augmented proliferation and motility of primary endometrial carcinoma cells. In vivo, oral administration of a single dose of IMT1 substantially inhibited endometrial carcinoma xenograft growth in the nude mice. mtDNA transcription inhibition, oxidative stress, ATP loss and apoptosis were detected in IMT1-treated endometrial carcinoma xenograft tissues. Together, targeting PLORMT by IMT1 inhibited endometrial carcinoma cell growth in vitro and in vivo.

Deep Learning for Intelligent Recognition and Prediction of Endometrial Cancer

The aim of the study was to investigate the intelligent recognition of radiomics based on the convolutional neural network (CNN) in predicting endometrial cancer (EC). In this study, 158 patients with EC in hospital were selected as the research objects and divided into a training group and a test group. All the patients underwent magnetic resonance imaging (MRI) before surgery. Based on the CNN, the imaging model of EC prediction was constructed according to the characteristics. Besides, the comprehensive prediction model was established through the clinical information and imaging parameters. The results showed that the area under the working characteristic curve (AUC) of the radiomics model and comprehensive prediction model was 0.897 and 0.913 in the training group, respectively. In addition, the AUC of the radiomics model was 0.889 in the test group and that of the comprehensive prediction model was 0.897. The comprehensive prediction model was established through specific imaging parameters and clinical pathological information, and its prediction performance was good, indicating that radiomics parameters could be applied as noninvasive markers to predict EC.

Immune Predictors of Radiotherapy Outcomes in Cervical Cancer

ABSTRACT The immune microenvironment influences the sensitivity of patients to radiotherapy (RT), yet determinants of therapeutic resistance remain elusive. This study integrates single‐cell transcriptomics and machine learning to delineate immune predictors of RT outcomes. Comprehensive analysis reveals reduced epithelial cell numbers, accompanied by enhanced apoptosis, complement activation, and inflammatory responses. RT triggers macrophage accumulation, particularly an RT‐responsive M1‐like HSPA1B+ subset with elevated antigen‐presenting capacity. While T and NK cell cytotoxicity increases, their exhaustion markers (e.g., PDCD1, TIGIT) are exacerbated. CellChat analysis identifies robust epithelial‐myeloid crosstalk mediated by the C3/C3AR1 axis. In murine models, C3AR1 antagonism diminishes RT efficacy, impairing macrophage infiltration and M1 polarization. Leveraging 25 single‐cell‐derived immune features, an 8‐feature multilayer perceptron model: Cervical Cancer Radiotherapy Immune‐Response Model (CCRTIM) is developed. CCRTIM robustly predicts prognosis (AUC = 0.76) and exhibits risk stratification. These findings unveil dynamic immune remodeling post‐RT and establish actionable biomarkers for precision radiotherapy strategies.

PRDM1-driven SLC30A9 overexpression contributes to the malignant phenotype of cervical cancer cells via promoting mitochondrial hyperfunction

Abstract Mitochondrial hyperfunction is important for cervical cancer progression. Solute carrier family 30 member 9 (SLC30A9) is a member of the solute carrier family 30, specifically a zinc transporter that plays a critical role in mitochondrial zinc homeostasis and maintaining mitochondrial function. We investigated the expression, function, and underlying mechanisms of SLC30A9 in the context of cervical cancer. Single-cell RNA sequencing analysis revealed a marked overexpression of SLC30A9 within the malignant epithelial cell population of cervical squamous cell carcinoma. This elevated SLC30A9 expression was further corroborated in clinical specimens from local patients and across various established and primary cervical cancer cells. SLC30A9 shRNA or knockout (via CRISPR/Cas9 method) significantly impeded the viability, proliferation, cell cycle progression and migration, and triggered apoptosis in cervical cancer cells. SLC30A9 depletion disrupted mitochondrial function, inhibiting mitochondrial respiration, mitochondrial membrane potential, mitochondrial complex I activity, and ATP production, also caused mitochondrial Zn 2+ accumulation, reactive oxygen species (ROS) production and oxidative injury. Conversely, overexpression of SLC30A9 in cervical cancer cells demonstrated enhanced mitochondrial complex I activity, increased ATP production, and augmented cellular proliferation and migration. Bioinformatic analysis, coupled with functional validation, identified PRDM1 (PR Domain Containing 1) as a key transcription factor regulating SLC30A9 expression. Silencing or knockout of PRDM1 resulted in a significant reduction in SLC30A9 promoter activity, as well as decreased SLC30A9 mRNA and protein levels in primary cervical cancer cells. Chromatin immunoprecipitation (ChIP) assays confirmed increased PRDM1 binding to the SLC30A9 promoter region in cervical cancer tissues. In vivo studies showed that SLC30A9 knockdown led to a remarkable decrease in the growth of xenografts formed by primary cervical cancer cells. These SLC30A9-silenced xenografts exhibited mitochondrial dysfunction, proliferation inhibition and apoptosis induction. These findings collectively suggest that PRDM1-driven SLC30A9 overexpression significantly contributes to the malignant phenotype of cervical cancer, possibly through promoting mitochondrial hyperfunction.

Prediction of Transcription Factors Binding Events Based on Epigenetic Modifications in Different Human Cells

E6 hijacks KDM5C/lnc_000231/miR‐497‐5p/CCNE1 axis to promote cervical cancer progression

Abstract Emerging evidence suggests that long non‐coding RNA (lncRNA) plays an important role in disease development, particularly in cancers. Recent studies with genome‐wide sequencing on cervical squamous cell carcinoma and matched adjacent non‐tumour tissues showed that a newly identified lncRNA‐lnc_000231 was highly expressed in cervical cancers. However, the underlying mechanism through which it is activated and its role in cervical cancer progression is still unclear. In this study, first, we confirmed that lnc_000231 is up‐regulated in cervical cancer cells and tumour tissues. Mechanically, we demonstrated that E6 up‐regulates lnc_000231 expression through promoting its promoter region H3K4me3 modification by destabilizing KDM5C. In vitro and in vivo results showed that lnc_000231 promotes cervical cancer cell proliferation and tumour formation by acting as miR‐497‐5p sponge and maintaining cyclin E1 (CCNE1) expression. Thus, our studies identified a new signalling pathway through which E6 promotes cervical cancer progression. E6 hijacked KDM5C/lnc_000231/miR‐497‐5p/CCNE1 signalling pathway is a promising target for cervical cancer treatment in the future.

Long noncoding RNA HCG11 inhibited growth and invasion in cervical cancer by sponging miR‐942‐5p and targeting GFI1

AbstractLong noncoding RNAs (lncRNAs) act as essential regulators in cancer tumorigenesis. Our study aimed to explore the underlying mechanism of lncRNA human leukocyte antigen complex group 11 (HCG11) in cervical cancer (CC) progression. Long noncoding RNA HCG11 was downregulated in CC. Functional assays demonstrated that lncRNA HCG11 inhibited CC cell proliferation and invasion. Then, we confirmed that lncRNA HCG11 could directly bind to miR‐942‐5p. Moreover, inhibition of miR‐942‐5p suppressed the growth and invasion of CC cells, and growth factor‐independent transcription repressor 1 (GFI1) gene was the target gene of miR‐942‐5p. Long noncoding RNA HCG11 increased the expression of GFI1 and suppressed cell proliferation and invasion by acting as a miR‐942‐5p sponge. Finally, the overexpression of lncRNA HCG11 suppressed the proliferation and metastasis of CC cells in vivo.

LINC00641 hinders the progression of cervical cancer by targeting miR‐378a‐3p/CPEB3

AbstractBackgroundLINC00641 was found to act in anti‐tumor manner in several types of cancers. Nonetheless, the detailed functions of LINC00641 have not been determined in cervical cancer (CC).MethodsThe expression of LINC00641, miR‐378a‐3p and CPEB3 was examined using a quantitative reverse transcriptase‐polymerase chain reaction. The relationships between LINC00641 and its downstream mechanism were illustrated by RNA pull‐down and luciferase reporter experiments.ResultsLINC00641 was found to be under‐expressed in CC cell lines. By overexpressing LINC00641, cell proliferative, migratory and invasive abilities, as well as epithelial mesenchymal transition (EMT) characteristics, were inhibited, whereas the rate of apoptosis was increased. Next, a starBase search (http://starbase.sysu.edu.cn) was applied to select microRNAs that had binding sequences with LINC00641. By up‐regulating LINC00641 expression, miR‐378a‐3p expression displayed the strongest decline. Moreover, miR‐378a‐3p was found to be up‐regulated in CC cell lines. In addition, LINC00641 hindered the progression of CC by decreasing miR‐378a‐3p expression. CPEB3 was discovered as a downstream target of miR‐378a‐3p and was under‐expressed in CC cells. Furthermore, knockdown of CPEB3 could counter the influence of an overexpression of LINC00641 with respect to CC progression.ConclusionsLINC00641 suppressed the progression of CC by targeting miR‐378a‐3p/CPEB3, suggesting that LINC00641 may have positive therapeutic impact for treatment for CC.

Utility of a Metabolic-Associated Nomogram to Predict the Recurrence-Free Survival of Stage I Cervical Cancer

MiR-1254 suppresses the proliferation and invasion of cervical cancer cells by modulating CD36

Abstract Background This study aimed to elucidate the roles of miR-1254 in cervical cancer progression and to explore the underlying mechanisms. Methods The expression levels of miR-1254 in normal-cancer cervical tissues and cells were measured using quantitive real-time polymerase chain reaction (qRT-PCR). The invasive and proliferative abilities of cervical cancer cell lines transfected with negative control (NC) mimic or miR-1254 mimic were measured using transwell, CCK-8, and colony formation assays. The binding sites between CD36 and miR-1254 were determined using luciferase reporter assays. The correlation of CD36 and miR-1254 with cervical cancer development was re-confirmed by co-transfection of miR-1254 mimic and CD36 overexpression using CCK-8, colony formation, transwell and western blot assays. Results MiR-1254 was expressed at significantly lower levels in the cervical cancer cell lines and tissues than in the controls. The functional assays revealed that upregulation of miR-1254 inhibited the invasion and proliferation of cervical cancer cells. The luciferase reporter assays demonstrated that CD36 messenger RNA and miR-1254 bound to one another. CD36 overexpression reversed the inhibitory effects of upregulated miR-1254 in the cervical cancer cells, suggesting that miR-1254 regulates cervical cancer progression by modulating CD36. Conclusion miR-1254 attenuated the invasion and proliferation of cervical cancer cells by modulating the expression levels of CD36.

Primary ewing sarcoma in the uterine cervix with multiple bone metastases

A 45-year-old female (G1, P1) with HPV-52 infection and low-grade cervical cytology presented to our clinic with an asymptomatic cervical mass. She suffered amenorrhea for the past 3 years due to medication for thyroid cancer. Magnetic resonance imaging revealed a mass sized 7.1 × 6.3 × 5.7 cm in the cervix with local invasion, and multiple lesions in the left ilium and femur. Histologically blue small round cells, positive for CD99 and FLI-1, obtained by cervical biopsy and the EWSR1 gene rearrangement detected by Fluorescence in situ hybridization confirmed the diagnosis of Ewing sarcoma. However, prior to the first cycle of systemic chemotherapy, the patient developed a sudden onset of lower extremities paraplegia, with PET/CT detecting extensive metastatic lesions in multiple bones including cervical 4 and thoracic 10 vertebral bodies. Although we performed emergency spinal canal decompression and excision of a thoracic 10 metastasis, the patient's condition deteriorated rapidly and died three months later.

Role of extracellular vesicle in human papillomavirus-associated cervical cancer

Cervical cancer is a gynecological malignant tumor and a serious threat to women's health. Although human papillomavirus (HPV) infection and the occurrence of cervical cancer are known to be closely related, the underlying carcinogenic mechanism of HPV is not fully understood. Extracellular vesicles (EVs) are found in a variety of body fluids and play an important role in both intercellular communication and cancer progression. Furthermore, the presence of EVs makes liquid biopsy of cervical cancer possible. The study of EVs in cervical cancer can provide clinical ideas for the diagnosis and treatment of the disease. The purpose of this article is to summarizes the role of EV contents in HPV-associated cervical cancer and discusses the possible clinical application of EVs in cervical cancer treatment. The search terms included the following: HPV with cervical cancer and extracellular vesicles. The initial literature search ended on March 1, 2023. In HPV-positive cervical cancer, EV contents are changed due to the presence of HPV. HPV-positive cervical cancer affects the cell microenvironment and other surrounding cells through the secretion of EVs.

USF1 regulated circPRDM4 modulates tumorigenesis and immune escape in chemoresistant cervical cancer

AbstractCervical cancer (CC) represents a major global health concern, characterized by chemoresistance and immune evasion mechanisms. Circular RNAs (circRNAs), which play a crucial role in cancer pathogenesis, particularly in the case of CC, have gained significant attention. The primary objective of this study was to investigate the functional significance of circRNAs in chemoresistant CC. A significant upregulation of circPRDM4 expression in chemoresistant CC cells. To investigate the functional consequences, we conducted circPRDM4 knockdown experiments, which resulted in the effective blockade of immune escape mechanisms employed by chemoresistant CC cells. Furthermore, circPRDM4 knockdown demonstrated a significant suppression of tumorigenesis in CC cells, highlighting its contribution to the oncogenic potential of CC. Investigating the regulatory mechanisms involved, we found that the transcriptional factor upstream stimulatory factor 1 (USF1) acts as an inducer of circPRDM4 expression. Remarkably, USF1 was found to effectively modulate CC cell immune escape via its interaction with circPRDM4. Moreover, our results revealed that USF1 is intricately involved in CC cell tumorigenesis through the regulation of circPRDM4. Collectively, our study elucidates the significant roles of circPRDM4 and its upstream regulator USF1 in chemoresistant CC cells. These findings underscore the importance of circRNAs in CC pathogenesis and provide valuable insights into the mechanisms underlying immune escape and tumorigenesis.