Epithelial-mesenchymal Transition (EMT) Generates Cells With Properties Of Cancer Stem Cells In Development And Progression Of Cervical Cancer

Background and objective:High-risk human papillomavirus (HPV) infection is the most important risk factor for cervical cancer, but studies have demonstrated that more than90%of HPV infections are cleared up without any medical action or consequences, only5%to10%will become persistent infection, and only the persistent infection can progress to cervical cancer. Therefore, high-risk HPV is necessary but not sufficient for the development of cervical cancer, so there are some other risk factors involved in addition to HR-HPV infection to developing a cervical cancer. However, the mechanism in this respect is not very clear. Understanding its mechanisms are conducive to the treatment plan, blocking the development of cancer, fundamentally inhibit the tumorogenesis of cervical cancer. Epithelial mesenchymal transition (EMT) and cancer stem cell plays a pivotal role in the mechanisms of epithelial cancer development. Recent studies have demonstrated that EMT plays a critical role not only in tumor metastasis but also in tumor recurrence and tumor initiating, which is tightly linked with the biology of cancer stem cells. This provides a new area for further study of the mechanisms of tumorigenesis. On this study, we would like to confirm that EMT induces the cells with stem cell properties in the malignant transformation of HPV infected cervical epithelial cells, to further clarify the pathogenesis of cervical cancer, improve and supplement the understanding of the pathogenesis of cervical cancer, develop a experimental and theoretical basis for early prevention and treatment of cervical cancer.Methods:1. Screening and identifing the TGF-β1and TWIST induced EMT in Ect, End and Siha cell lines. Ect, End and Siha cells will be stimulated by TGF-β1. To build the pcDNA4.0-TWIST vector and screening the stable expression of the TWIST in End and Siha cells. Expression changes of the EMT-related markers will be detected after induction of the EMT.2. Detecting the stem cell properties in EMT induced cervical epithelial and cancer cells Detect the expression changes of stem cell related markers in EMT induced End and Siha cells on mRNA and protein level.3. Functional validation of the cervical cancer stem cells in vitro Validate the self-renewal and differentiation potential capacity of EMT induced stem cells by Stem Cell Colony Formation Assay and ck10ck13immunofluorescence detection experiments.Results:1. EMT regulatory transcription factor TGF-beta1and TWIST were successful induced EMT changes in the HPV immortalized cervical epithelial cells and cervical cancer cells.2. TGF-beta1and TWIST induced EMT changes generates the cervical epithelial cells and cervical cancer cells with the properties of stem cell.3. In Vitro validation confirmed that EMT induced stem cells in cervical epithelial and cervical cancer cells have the characteristics of self-renewal and multi-differentiation potential capacity.Conclusion:EMT key regulatory factor TGF-β1and TWIST induced cervical epithelial and cervical cancer cells with the properties of stem cells which have self-renewal and differentiation potential characteristics, which may induce the malignance behavior of cervical cancer. Background:Small sample size used in previous studies results in lack of overlap between the reported gene signatures for prediction of chemotherapy response. Recently, The Cancer Genome Atlas (TCGA) research-network concluded an ovarian-cancer study and released the dataset to the public. The TCGA dataset possesses large sample size, comprehensive molecular profiles, and clinical outcome information. In this study, we have used a large population of patients to identify copy number variation and gene expression signatures associated with chemotherapy response in serous ovarian carcinoma.Methods and Results:The gene expression and copy number profiles of569tumor samples from the TCGA ovarian cancer dataset are analyzed using a semi-supervised clustering method combined with a novel scoring function. A gene regulation network was then built to further identify genes responsible for differential gene expression and copy number variation between the chemotherapy resistant group and the chemotherapy sensitive group. We have identified460genes with significant deletion or amplifications among the chemo-resistant group. Utilizing associated gene expression data we predict94genes with altered copy number and correlated changes in expression. Among them, ten important genes and six critical signaling pathways were identified to have important implications in chemotherapeutic response. Further, ten potential serum biomarkers were identified for predicting chemoresistance in serous ovarian carcinoma.Conclusions:A gene signature discovered on a large data set provides robustness in accurately predicting chemotherapy response in serous ovarian carcinoma. The combination of copy number variation and gene expression signatures help to understanding of potential mechanisms involved in drug resistance. LIGHT is a tumor necrosis factor (TNF) superfamily ligand which is considered as a promising candidate for cancer therapy; It has a potent anti-tumor activity through establishing lymphoid-like tissues inside tumor sites and recruiting naive T cells into the tumor. In this study, we examined the possibility of antitumor activity by expressing LIGHT in cervical cancer model. A recombinant adeno-associated virus (rAAV) vector was chosen for the transfer, based on its transfection efficiency and lack of detectable pathology. In vitro transfer of rAAV vector expressing LIGHT (AAV-LIGHT) stimulated T-lymphocyte proliferation and activation. Adeno-associated virus mediated gene transfer of LIGHT by intratumoral injection exerted a very potent antitumor effect against pre-existing TC-1cell cervical cancer in C57BL/6mice. This study confirmed that AAV-LIGHT regressed tumor growth by activating cytotoxic T lymphocyte (CTL), enhancing infiltration of inflammatory cells in tumor and increasing stimulatory cytokine expression in tumor microenvironment. Therefore, AAV-LIGHT therapy might have potential utility for the treatment of cervical cancer.