Journal of Reproductive Immunology

Possible roles of HLA-G regulating immune cells in pregnancy and endometrial diseases via KIR2DL4

Major histocompatibility complex (MHC) molecules bind peptides originated from cellular synthesis and present them at the cell surface for recognition by receptors on immune cells like T lymphocytes, natural killer (NK) cells or mast cells. Such recognition plays a crucial part in autoimmunity, anti-bacterial, anti-viral and anti-tumor responses. Human leukocyte antigen G (HLA-G) is a member of non-classical HLA class I molecules which has been studied deeply in recent years into its role in pregnancy and endometrial diseases, including endometrial tumor, endometriosis and adenomyosis, etc. Understanding the mechanism of the maintenance of pregnancy and immune escape of endometrial diseases in a HLA-G dependent way is of current interest. Perception from studies in the expression of HLA-G and possible pathways is a vital part of understanding mechanisms related to immune escape.

RETRACTED: Differential expression of miR-130a-3p modulate ovarian epithelial carcinoma (OEC) cell development and could be a biomarker for OEC

A lot of research investigations, conducted in the recent years, establishes that microRNAs (miRNAs) have an important role in keeping both growth and metastasis of Ovarian Epithelial Carcinoma (OEC) under control. However, the clinical and functional role of miR-130a-3p in OEC is yet to be explored. Through quantitative reaction i.e., qRT-PCR, the expression of miR-130a-3p was assessed in tissues and cell lines of OEC patients. This analysis determined the relationship between the expression of miR-130a-3p and its clinicopathology with the overall survival rate of OEC patients. The author made use of cell counting analysis (CCK8) and in vitro flow cytometry to understand the functional and biological impact of miR-130a-3p expression. In comparison with neighboring normal tissues, the expression of miR-130a-3p was found to be lower in 7 OEC samples. Few reasons are cited for this scenario i.e., low expression of miR-130a-3p, such as low overall OEC patient survival rate, incidence of FIGO and metastasis of lymph nodes. miR-130a-3p has been found as an independent candidate for predicting the prognosis of OEC patients, as per Multivariate Cox research. When miR-130a-3p is over-expressed, as per the enhanced mechanism, it prevents both cell proliferation and cell cycle production in OEC. The current study findings emphasize that miR-130a-3p can be leveraged as a biomarker of prognosis and possibly as a target in the treatment of OEC.

RETRACTED: MiR-374b increases the CIK expression and mediates biological function changes in cervical cancer cells by targeting the PD-1/PD-L1 signaling pathway

To investigate the role of miR-374b in medicating biological function changes in cervical cancer cells by increasing the cytokine-induced killer (CIK) expression. Venous blood of 62 cervical cancer patients and 58 healthy individuals including Human cervical cancer cell line (HeLa) and normal human uterine smooth muscle cells (HUSMC) were tested for expression of miR-374b, PD-1, and PD-L1. sh-PD-1, si-PD-1, NC, miR-374b-inhibitor, and miR-374b-mimics were transfected into HeLa cells, and expression of miR-374b, PD-1, and PD-L1 was determined by a qRT-PCR assay, and the proliferation and apoptosis of the cells were detected using a MTT assay and flow cytometry, respectively. PD-1 was highly expressed in cervical cancer, while miR-374b is lowly expressed in it, and the area-under-the-curve (AUC) of both PD-1 and miR-374b was larger than 0.8. The dual luciferase reporter assay confirmed relationship between PD-1 and miR-374b. Expression of PD-1 in HeLa cells was significantly down-regulated after transfection of miR-374b-mimics. Compared with the CIK + NC group, the CIK + miR-374b-mimics group and the CIK + siRNA-PD-1 group showed a significant decrease in the relative mRNA expression of PD-1, compared with other group showed significantly lowered activity of HeLa cells, and the two groups showed significantly reduced tumor volume. MiR-374b increases the CIK expression and mediates biological function changes in cervical cancer cells by targeting the PD-1/PD-L1 signaling pathway, so it is expected to be a potential therapeutic target for cervical cancer.

RETRACTED: G-5555 synergized miR-485-5p to alleviate cisplatin resistance in ovarian cancer cells via Pi3k/Akt signaling pathway

The present study was meant for the discovery of the underlying functions of miR-485-5p in ovarian cancer concerning cisplatin resistance in vitro. RT-qPCR assessed the miR-485-5p expression in ovarian cancer cell lines, normal cells and cisplatin-resistant Cell line OVCA433-CR. After OVCA433-CR treated with 0,3,5umol/L cisplatin, miR-485-5p expressions were determined. MTT observed the cell cytotoxicity in OVCA433-CR after regulation of miR-485-5p. Targets can predicted the putative binding between miR-485-5p and PAK1 and Luciferase Assay verified this. RT-qPCR decided the inhibitory effect in between. MTT tested the cytotoxicity in different combinations of miR-485-5p and PAK1. Western Blot tested the phosphorylation of Pi3k and Akt in response to miR-485-5p and PAK1 interplay. We evaluated the role of Pi3k/Akt signaling in regulation of miR-485-5p and cisplatin resistance with Wortmannin. miR-485-5p was lower expressed in ovarian cancer cells than normal ones and even lower in OVCA433-CR than OVCA433. As the cisplatin concerntration increased, miR-485-5p decreased. miR-485-5p mimics induced lower cisplatin resistance while miR-485-5p inhibitor caused higher resistance. PAK1 targeted miR-485-5p and inhibited miR-485-5p. PAK1 inhibitor helped to lower the resistance to cisplatin caused by miR-485-5p upregulation. miR-485-5p mimics silenced Pi3k/Akt signaling and PAK1 inhibitor aggravated the silencing. Inhibition of Pi3k/Akt signaling increased miR-485-5p, thereby decreasing the cisplatin-resistance in OVCA433-CR. miR-485-5p decreased cisplatin resistance in ovarian cancer cells via Pi3k/Akt signaling, suggesting that miR-485-5p upregulation might alleviate the cisplatin resistance in ovarian patients.

Exosome's role in ovarian disease pathogenesis and therapy; Focus on ovarian cancer and failure

In the eukaryotic system, exosomes are categorized as unique extracellular vesicles with dimensions ranging from 30 to 150 nm. These vesicles contain a variety of endogenous molecules, such as proteins, DNA, mRNA, microRNA, and circular RNA. They are essential for a wide range of metabolic events and have the potential to be used as therapeutic or diagnostic targets for a number of diseases, including ovarian diseases. By inducing changes in the surrounding environment, the donor exosomes transfer their contents to the receiving cells, so demonstrating the biological implications of major interactions between cells. Mesenchymal stem cells (MSCs) have produced exosomes have shown promise as a treatment for premature organ failure (POF or POI). Furthermore, exosomal transport has many complexities, and contributes to the pathophysiology of ovarian cancer by affecting cell growth, migration, metastastsis and etc. Owing to these facts, in this paper, we present the progress developed in the understanding of exosomes as a viable therapeutic avenue and indisputable prognostic targets in ovarian disorders.

Potential tools for predicting response to chemotherapy in OC: Assessment of immune dysbiosis, participant’s self-rated health and microbial dynamics

Epithelial ovarian cancer (OC) is the deadliest female reproductive cancer; an estimated 13,270 women will die from OC in 2023. Platinum-based chemotherapy resistance mechanisms contribute to poor OC 5-year survival rates. Peripheral inflammation is linked to various disease states and we previously identified unique peritoneal microbial features predictive of OC. We hypothesized that unique peripheral immune profiles and peritoneal microbial features may be predictive of disease-free interval (time to recurrence) and response to chemotherapy in participants with OC. We also investigated self-rated health (SRH) scores in the context of peripheral inflammation as a potential screening tool for OC. Blood and peritoneal fluid were collected from participants with OC or a benign adnexal mass (BPM). Lymphocyte populations were analyzed using Fluorescence Activated Cell Sorting, serum cytokine levels were analyzed using the Human Th17 Magnetic Bead Panel assay and peritoneal fluid microbial features were analyzed using Next Generation Sequencing (NGS). Participants completed a standardized questionnaire on self-rated physical and emotional health. Participants were classified into three chemotherapy response categories: platinum-refractory, platinum-resistant or platinum-sensitive. A significant positive correlation was found between elevated inflammatory status on the day of surgery and longer disease-free interval. SRH measures did not correlate with immune status in participants with OC or a BPM. We identified a correlation between peritoneal microbial features and chemotherapy response. We conclude that immune dysbiosis may be useful in predicting OC recurrence. The immune findings reported here set the framework for additional studies utilizing immune profiles to predict platinum-based chemotherapy responsiveness in OC.

HLA-G - evolvement from a trophoblast specific marker to a checkpoint molecule in cancer, a narrative review about the specific role in breast- and gynecological cancer

Combination of IDO1high and CCL19low expression in the tumor tissue reduces survival in HPV positive cervical cancer

The over expression of Indoleamine 2, 3-Dioxygenase (IDO1), an immune checkpoint inhibitor, is well known in cervical cancer. However, its association with chemokine signals promoting cellular infiltration in the cervical tumor microenvironment, is unknown. In the current study, we evaluated the expression and enzymatic activity of IDO1. We also profiled the expression of chemokine ligand-receptors- CCR4-CCL22, CXCR3-CXCL10, CXCR4-CXCL12, and CCR7-CCL19 using immunohistochemistry (IHC), and studied their association with IDO1, statistically. After getting an informed consent, punch biopsy samples were obtained from 105 patients diagnosed with cervical cancer. HPV typing by Sanger sequencing, realtime PCR for quantifying IDO1 mRNA expression, HPLC for determining the K/T ratio and IHC for all the above chemokine receptor-ligand pairs along with IDO1 were performed. We found a significant increase in the expression of IDO1 and K/T levels in early and locally advanced stages when compared to Stage IV disease. Among the chemokine ligand -receptor pairs profiled, we found that high CCL19 marker expression was a good prognostic indicator of patients' disease-free (p = 0.013) and overall survival (p = 0.043). Although we could not identify IDO1 as an independent prognostic factor, we found that high levels of IDO1 expression may further reduce survival outcomes in patients with low CCL19 expression. This could be vital for designing immuno therapeutic interventions targeting IDO1.

Promising anti- cervical carcinoma and inflammatory agent, Resveratrol targets poly (ADP-ribose) polymerase 1 (PARP-1) induced premature ovarian failure with a potent enzymatic modulatory activity

Radioprotective effects of Resveratrol is well known in normal cells exposed to the damaging effects of ionizing radiation however, its potential radioprotective effect on ovarian follicle formation and development is still uncertain. Astonishingly, it has been reported that PARP contributed to the pathogenesis of immune-mediated ovarian injury. In this paper, Resveratrol was tested for its inflammatory, anti-cervical carcinoma activity, and checked its targets poly (ADP-ribose) polymerase 1 (PARP-1) induced premature ovarian failure with a potent enzymatic modulatory activity. Through high-throughput virtual screening method, Resveratrol was screened to find its target. That the compound strongly inhibited cervical carcinoma HT-3 cell. The cell proliferation was evaluated by an CCK-8 assay, and the cell apoptosis was assessed by a flow cytometry. Rat model of premature ovarian failure was used to introduce resveratrol preparation and rtPCR was done to measure expression of apoptosis related markers. We report resveratrol as a potential target for PARP-1 and its modulator from a high-throughput virtual screening method. Resveratrol was measured its anti-cervical carcinoma activity by using an CCK-8 assay, which suggested that the compound strongly inhibited HT-3 cell proliferation, the IC

Moesin affects the plasma membrane expression and the immune checkpoint function of CD47 in human ovarian clear cell carcinoma

Among major histological subtypes of epithelial ovarian cancer, a higher incidence of ovarian clear cell carcinoma (OCCC) is observed in East Asian populations, particularly in Japan. Despite recent progress in the immune checkpoint inhibitors for a wide variety of cancer cell types, patients with OCCC exhibit considerably low response rates to these drugs. Hence, urgent efforts are needed to develop a novel immunotherapeutic approach for OCCC. CD47, a transmembrane protein, is overexpressed in almost all cancer cells and disrupts macrophage phagocytic activity in cancer cells. Ezrin-Radixin-Moesin (ERM) family member of proteins serve as scaffold proteins by crosslinking certain transmembrane proteins with the actin cytoskeleton, contributing to their plasma membrane localization. Here, we examined the role of ERM family in the plasma membrane localization and functionality of CD47 in OCCC cell lines derived from Japanese women. Confocal laser scanning microscopy analysis showed colocalization of CD47 with all three ERM in the plasma membrane of OCCC cells. RNA interference-mediated gene silencing of moesin, but not others, decreased the plasma membrane expression and immune checkpoint function of CD47, as determined by flow cytometry and in vitro phagocytosis assay using human macrophage-like cells, respectively. Interestingly, clinical database analysis indicated that moesin expression in OCCC was higher than that in other histological subtypes of ovarian cancers, and the expression of CD47 and moesin increased with the cancer stage. In conclusion, moesin is overexpressed in OCCC and may be the predominant scaffold protein responsible for CD47 plasma membrane localization and function in OCCC.

LPS-induced TLR4 activation enhances NF-κB signalling in human ovarian granulosa-like tumor cell line and induced primordial Follicle activation in mouse ex vivo ovary assay

The immune system is increasingly recognized as a regulator of ovarian function, yet its role in early folliculogenesis remains unclear. Ovulation exhibits inflammatory-like processes involving Toll-like receptors, which is expressed in early follicles and may influence their activation. This study aims to demonstrate how the TLR4 inhibitor C34 reduces primordial follicle activation in an ex vivo ovary assay. We further examine how this effect relates to LPS stimulation and its interaction with the well-known primordial follicle activation pathway, the AKT signaling pathway. Using Western blotting, we observe increased phosphorylation of AKT. In KGN cells, LPC-induced TLR4 activation triggers NF-κB signalling and ELISA reveals production of the monocyte chemoattractant protein 1 (MCP-1). MCP-1 enhances AKT phosphorylation and cell proliferation, suggesting a link between immune signalling and follicle activation. These findings indicate that LPS-induced activation of TLR4 may interact with PI3K-AKT signalling to regulate primordial follicle activation and ovarian reserve maintenance.

SQSTM1/p62 drives radiosensitivity in cervical cancer via autophagy-independent mechanisms

Cervical cancer comprises squamous cell carcinomas (SCCs), which are generally radiosensitive; meanwhile, adenocarcinomas respond poorly to radiation. Here, we explored the role of selective autophagy receptors in modulating radiosensitivity across these subtypes. We found that SQSTM1/p62 was highly expressed in human papillomavirus (HPV)-positive SCC cell lines (HeLa, ME180) and clinical SCC specimens, but was low or undetectable in HPV-negative adenocarcinomas and the C33A cell line. Clonogenic assays confirmed that HPV-positive cells exhibited greater radiosensitivity than C33A. Upon irradiation, only HPV-positive cells showed upregulation of p62 and DNA-damage response proteins, whereas C33A cells did not. Treatment with autophagy inhibitors (bafilomycin A1 and chloroquine) further increased p62 levels in HPV-positive cells but not in HPV-negative cells. The sustained low p62 levels in C33A cells, regardless of autophagy inhibition, may represent a distinctive biomolecular feature of HPV-negative cervical cancer. Notably, overexpression of Atg4B to block autophagic flux without affecting p62 expression had no impact on radiosensitivity in either cell type, whereas pharmacologic inhibition of autophagy selectively enhanced radiation-induced cytotoxicity in HPV-positive cells. Conversely, siRNA-mediated knockdown of p62 in HPV-positive cells attenuated radiation-induced growth suppression. Together, these data indicate that p62 promotes radiosensitivity in HPV-positive cervical cancer through a mechanism distinct from classical autophagy. Therefore, p62 represents both a predictive biomarker for radiation response and a potential target for radiosensitization strategies in HPV-positive tumors, while its unresponsiveness in HPV-negative cancers suggests the need for alternative approaches in that context.

MiR-195-3p inhibits cell proliferation in cervical cancer by targeting BCDIN3D

Cervical cancer (CC) is one of the most prevailing cancers among females. Accumulated studies concentrated on the regulatory role of micro RNA in cancers. This research is to explore the potential role of mir-195-3p in cervical cancer progression. Bioinformatics tools were used to investigate differential expression of mir-195-3p and BCDIN3D in cervical cancer. RNA expression patterns of both mir-195-3p and BCDIN3D were detected by RT-PCR in CC cell lines. The protein expression of BCDIN3D was measured by Western Blot. Hela and Siha cell lines were transfected with mir-195-3p inhibitors, mir-195-3p mimics and BCDIN3D si-RNA, si-NC. Luciferase reporter assays were adopted to confirm the binding. The interplays between mir-195-3p and BCDIN3D were explored in CC cell lines. CCK-8 assays checked how mir-195-3p regulated cell proliferation and Ki67 was examined by Western blot for its protein expressions as a biomarker for CC cell proliferation. MiR-195-3p was downregulated while BCDIN3D was upregulated in cervical cancer cell lines. The binding was confirmed via Luciferase Assay. RT-PCR suggested that upregulation of mir-195-3p inhibited BCDIN3D and downregulation of BCDIN3D in return induced mir-195-3p. CCK-8 pointed out that overexpression of mir-195-3p inhibited the cell viability. Ki67 protein expression was inhibited by miR-195-3p mimics or silence of BCDIN3D. The present research led us to a conclusion that mir-195-3p might inhibit cervical cancer cell proliferation and was reversely regulated by BCDIN3D. This suggests that miR-195-3p mimics/ BCDIN3D si-RNA might be used in the treatments of cervical cancer in the future after various animal assays and clinical trials.

Role of inflammation and immune response in the pathogenesis of uterine fibroids: Including their negative impact on reproductive outcomes

Uterine fibroids (UFs), the most common tumors in women of reproductive age, are characterized by sex steroid-dependent growth and excessive deposition of extracellular matrix (ECM) surrounding UF smooth muscle cells. Women with symptomatic UFs experience heavy menstrual bleeding and consequent iron-deficiency anemia. They also have a risk of recurrent pregnancy loss, preterm birth, and cesarean delivery, indicating that UFs can negatively affect reproductive outcomes. Various types of immune cells, including innate and adaptive cells, are observed in UFs; however, the impact of these cells on the pathophysiology of UFs remains unclear. Inflammation may play important roles in the growth of UFs, and expression levels of proinflammatory and inflammatory cytokines, such as interleukin (IL)-1, IL-6, IL-10, TNF-α, and TGF-β, are upregulated in UFs. These cytokines play important roles in the interaction between growth factors and ECM that is regulated by the sex steroids estrogen and progesterone. Furthermore, proinflammatory mediators are upregulated in females with UFs, with higher expression levels in the endometrium with submucosal and intramural UFs than in the endometrium without UFs, indicating that these proinflammatory cytokines may impair endometrial receptivity, leading to implantation failure in in vitro fertilization programs. Hormonal treatments using gonadotropin releasing hormone analogs and the selective progesterone receptor modulator ulipristal acetate significantly shrink UFs and improve UF-related symptoms. These compounds can regulate local inflammation in UFs and adjacent myometrium. Controlling and improving local inflammation caused by UFs may represent a novel therapeutic strategy for UFs and potentially improve reproductive outcomes in women with symptomatic UFs.

Radixin modulates the plasma membrane localization of CD47 in human uterine cervical adenocarcinoma cells

Despite the dramatic success of immune checkpoint blockers in treating numerous cancer cell types, current therapeutic modalities provide clinical benefits to a subset of patients with cervical cancers. CD47 is commonly overexpressed in a broad variety of cancer cells, correlates with poor clinical prognosis, and acts as a dominant macrophage checkpoint by interacting with receptors expressed on macrophages. It allows cancer cells to escape from the innate immune system and hence is a potential therapeutic target for developing novel macrophage checkpoint blockade immunotherapies. As the intracellular scaffold proteins, ezrin/radixin/moesin (ERM) family proteins post-translationally regulate the cellular membrane localization of numerous transmembrane proteins, by crosslinking them with the actin cytoskeleton. We demonstrated that radixin modulates the plasma membrane localization and functionality of CD47 in HeLa cells. Immunofluorescence analysis and co-immunoprecipitation assay using anti-CD47 antibody showed the colocalization of CD47 and all three ERM families in the plasma membrane, and the molecular interactions between CD47 and all three ERM. Interestingly, gene silencing of only radixin, reduced the CD47 plasma membrane localization and functionality by means of flow cytometry and phagocytosis assay but had little influence on its mRNA expression. Together, in HeLa cells radixin may function as a principal scaffold protein responsible for the CD47 plasma membrane localization.

MiR-624-5p enhances NLRP3 augmented gemcitabine resistance via EMT/IL-1β/Wnt/β-catenin signaling pathway in ovarian cancer

The current study investigates the NLRP3's cytotoxicity inhibitory effect among ovarian cancer cells and how it interacts with Wnt/β-catenin in vitro conditions. Further, the study also analyzed the regulatory role of NLRP3 in resistance to gemcitabine among ovarian cancer cells and its underlying interaction mechanisms with Wnt/β-catenin in vitro. The current in vitro study detailed that when downregulating NLRP3, it could enhance the gemcitabine sensitivity in GRC cells. In case of gemcitabine-resistant cells, the up-regulation of NLRP3 can increase the drug-resistance through the activation of IL-1β, EMT and Wnt/β-catenin signaling pathways. High expression of miR-624-5p was recorded in ovarian drug resistant cancer cells and it also boosted the cell viabilities. NLRP3 can reinstate the functioning of miR-624-5p in drug resistant cells. This phenomenon concludes that NLRP3 is a promising therapeutic target and can be implemented in traditional chemotherapy to increase the efficacy of the treatment. The current study conducted in vitro experiments and the findings infer that the downregulation of NLRP3 can enhance the sensitivity of gemcitabine among GRC cells. This mechanism will increase the treatment efficacy by inhibiting the drug resistance in GRC. These two entities are the new promising biomarkers that can be used in the detection of platinum resistance in ovarian cancer patients and conduct novel clinical research.

Understanding HLA-G driven journey from HPV infection to cancer cervix: Adding missing pieces to the jigsaw puzzle

Human Papillomavirus (HPV) is a vital risk-factor for cancer cervix. However, persistent HPV infection results in cervical cancer in only a minority. Probably, HPV subdues the host immune response for persistence, which includes augmentation of HLA-G and plausibly aids in progression to cervical cancer. HLA-G, which comprises of membrane and soluble form, downregulates the host's immune response and generate tolerance. The current study aimed to analyze both forms of HLA-G in fresh tissue and plasma of women with HPV-infected and uninfected cervix and cancer cervix using Western blot and ELISA. The study cohort included 30 women with cervical carcinoma and equal number with normal cervix and 6 with HPV infected cervix. We observed a significant upregulation of membranous HLA-G expression in HPV infected cervix and cervical carcinoma (P < 0.001). Interestingly, the pairwise comparison of HLA-G tissue protein expression of the normal cervix and cervical carcinoma, as well as the normal cervix with HPV infected cervix, was significant (P < 0.001). Levels of soluble HLA-G were significantly raised in carcinoma cervix. We observed a progressive increase in HLA-G protein expression in HPV infected cervix and cervical carcinoma. These findings compel us to hypothesize that the upregulation of HLA-G expression favors the persistence of HPV in a microenvironment of a submissive host response. This progressive upregulation further leads to cervical cancer. Thus elimination of HPV infection seems to be a desirable proposition to prevent cervical cancer. In the absence of antiviral therapy for HPV, exploration of HLA-G antibody-based therapeutic strategies appear promising.

RETRACTED: Interplay of microRNAs to genetic, epigenetic, copy number variations of cervical cancer related genes

This study was to analyses the miRNAs role in cervical cancer and possibilities of microRNA-based markers as diagnostic tools. Genome wide analysis was performed for CNV detection using PennCNV and QuantiSNP. The associated mRNA qRT-PCR detection was used to measure quantities of microRNA gene expression. More than 10 CNV regions has a significant relationship with cervical cancer risk for both CNV detection algorithms. A total of 34 CNVs was detected by QuantiSNP while it was 27 in case of PennCNV, among which 22 CNVs was found to be overlapping between these two algorithms. the mRNA was analyzed for its expression on 36 carvical tumor normal tissue pairs of four targets i.e., MAP3K3, RIPK2, DIRAS3 and GAS7. These infers that there was a significant downregulation of all the four genes cervical tumor. Our results showed that miR-182 can modulate the expression of FAM83H, DIRAS3, RIPK2 and MAP3K3 in cervical cancer. Therefore, indicated that miR-182 can acts through these signaling pathway in proliferation of cervical cancer cells. The expression of tumor modulator miRNAs can be controlled by miRNA replacement therapy. Several miRNAs have been used for this purpose. The modulation of various signaling pathway and proteins in cervical cancer cells by miR-182 needs further clarification.

Activating transcription factor 6 regulated cell growth, migration and inhibiteds cell apoptosis and autophagy via MAPK pathway in cervical cancer

Cervical cancer cell function is influence by ER. Therefore, in this study, ER stress senser-ATF6, was selected for detailed research in cervical cancer. ATF6 mRNA was assessed through RT-qPCR assays. Cell transfection was to regulate ATF6 and thereafter the differential ATF6 cancer cells were divided into two groups for further functional assays. Cell viabilities were analyzed by CCK-8 and migration by Scratch. RT-qPCR examined cell death biomarkers Caspas-3 and Bcl-2. 4-PBA was utilized to inhibit ER stress. After that, ATF6, viability, migration and apoptotic proteins were scrutinized after ER inhibition. Proteins signifying EMT, autophagy and MAPK signaling pathway were checked by western bolt. Last, we inactivated the MAPK signaling to investigate into the changes in cell functions. ATF6 presented higher expression in cervical cancer cells. Inhibited ATF6 could reduce cell viabilities and migration but promote apoptosis through suppressing Bcl-2 and increasing caspase-3. ER stress antagonist witnessed a drop in ATF6 expression, cell viability, migration and Bcl-2 but a rise in caspase-3 activation, suggesting apoptosis increase. Cell autophagy was hindered in CC cells. Knockdown of ATF6 promoted autophagy and restrained EMT and MAPK signaling pathway. Suppressed ERK1/2 obstructed cell viabilities, migration, EMT and autophagy but promoted apoptosis. ATF6 might promote cell growth, migration, autophagy through ER stress and MAPK signaling in cervical cancer in vitro, indicating a potential regulatory gene in cervical cancer. However, in-depth researches are requested to enrich the knowledge of ATF6 in cervical cancer in vivo and in clinical in the future.

Killing cervical cancer cells by specific chimeric antigen receptor-modified T cells

The aim is to investigate the in vivo and in vitro killing effect of mesothelin chimeric antigen receptor T cells (MESO-CAR-T) in cervical squamous cell carcinoma. MESO-CAR-T cells were successfully constructed. In vitro verification of the killing effect of MESO-CAR-T cells was evaluated in the presence of SiHa cells by the lactate dehydrogenase release assay and cytokine release assay. The in vivo experiments were performed in immunodeficient NCG mice. After successful tumor formation with the subcutaneous implantation of SiHa cervical cancer cells, the injections of MESO-CAR-T cells into the tumors at different doses and frequencies were performed. Subsequently, the growth rate and size of the tumors in NCG mice were observed. A 17-fold increase in the number of MESO-CAR-T cells and a 16-fold increase in the number of Con-CAR-T cells were observed. The result of marker detection in the prepared MESO-CAR-T cells showed that CD3

Frequent expression of CD45RO memory T cell marker as well as low to high expression of PD-1 and PD-L1 inhibitory molecules in seminoma and dysgerminoma

Evaluation of the genetic diversity of KIR genes in patients with endometrial cancer

Endometrial cancer (EC) constitutes more than half of all genital cancers in women, with an increasing incidence in different countries. Natural killer cells (NK cells) are kinds of innate immune cells that are controlled by sets of receptors, such as killer cell Ig-like receptors (KIRs), which can inhibit or activate NK cells. In this study, we evaluated the diversity and genetic association of KIRs in confirmed cases of endometrial cancer compared to healthy controls. A total of 151 women with EC and 167 age/race-matched healthy controls were analyzed for KIR genes. Demographic and histopathologic data were gathered in questionnaires, and 16 KIR genes along with two variants of KIR2DS4 (KIR2DS4fl and KIR2DS4del), were genotyped by usingsequence specific primers-polymerase chain reaction (SSP-PCR) method. A comparison between cases and controls revealed that although there were not any significant differences in A haplotype associated genes and also the variants of KIR2DS4 (p >0.05), B haplotype associated genes such as KIR2DS2 and KIR2DL2 decreased significantly in EC patients in comparison with healthy controls (p=0.03 and p=0.01, respectively). Furthermore, we found that EC mostly developed in cases with the AA genotype; however, the carriers of Bx and C4T4 genotypes were less frequent in patients with EC. Our results revealed that KIR2DS2 and KIR2DL2, along with Bx and C4T4 genotypes, have a protective impact against developing endometrial cancer in Iranians.

Upregulation of B3GNT3 is associated with immune infiltration and activation of NF-κB pathway in gynecologic cancers

Beta-1,3-N-acetylglucosaminyltransferase-3 (B3GNT3) is a carcinogenic factor in many cancers. However, the biological functions of B3GNT3 in gynecologic cancers especially in cervical, ovarian, and endometrial cancers are mostly limited. B3GNT3 levels in pan-cancer and gynecologic cancers were predicted by GSCA, GEPIA, and the Human Protein Atlas databases. The overall survival was predicted by Kaplan-Meier Plotter. ROC curve was generated according to the TCGA database. The immune cell infiltration was analyzed by ImmuCellAI algorithm using GSCA database. The related genes and pathways were analyzed via LinkedOmics portal, GO, KEGG pathway enrichment analysis, and GEPIA database. B3GNT3, p-p65/p65, and nuclear p65 levels were detected by western blotting. B3GNT3 was differentially expressed in pan-cancers. B3GNT3 expression was increased in cervical, ovarian, and endometrial cancers. High expression of B3GNT3 was associated with lower overall survival, and was used for diagnosis of these gynecologic cancers. B3GNT3 was related to different immune cell infiltration. B3GNT3 was associated with NF-κB signaling activation by regulating multiple related biomarkers. Silencing of B3GNT3 repressed activation of the NF-κB signaling in gynecologic cancers. In conclusion, upregulated B3GNT3 is related to diagnosis, poor prognosis, immune infiltration, and activation of the NF-κB signaling in gynecologic cancers.

The roles and therapeutic applications of cytokines in endometrial cancer

Endometrial cancer (EC) is a common gynecological cancer globally and the most frequent gynecologic malignancy in industrialized countries. Patients are typically diagnosed when the disease is still restricted to the uterus. 5-year overall survival ranges from 70 % to 90 % in patients without metastatic disease; however, the metastatic form of the disease affects 16 % of EC patients, with a 5-year survival rate of 16.8 %. The immune system can detect abnormal cells as non-self in the early stages of carcinogenesis, producing the appropriate pro-inflammatory environment to eliminate cancer cells. In a second phase, cancer cells use various immune-editing systems to alter the profile of the immune response from pro to anti-inflammatory, resulting in immune escape. The directors of this immune switching mechanism are cytokines. Studies have reported the increased expression of several pro-and anti-inflammatory cytokines in EC tissues and cell lines, including Interleukin (IL)- 6, IL-8, IL-31, IL-33, IL-10, TGF-β, VEGF, and IL-1Ra. Immune cells producing these cytokines have also been reported to be present in EC tissues. Therefore, in this study, we aimed to show the possible mechanisms of the mentioned cytokines on EC progression, as well as the most current and prospective advancements in cytokine-based therapeutic applications.