Cxcl1 Gene In The Biological Effects Of Ovarian Malignancy

Chapter 1 The study of the biological effects of CXCL1 gene in ovarian cancerOvarian cancer is the gynecologic malignancy in the incidence of third place in the mortality rate ranks,but No.1 in the maliganat disease.As the ovary in pelvic anatomy on the hidden depths of the early onset of the lack of specific symptoms and signs, difficulty in early detection and diagnosis, about 60%-70% of ovarian cancer when patients are diagnosed late. The past 30 years, gynecologic oncology experts around the world despite the tireless efforts to continuously improve the surgical procedure and found that newer and better chemotherapeutic drugs, the addition of new chemotherapy regimens, but the 5-year survival rate has hovered around 20%-30%. Therefore, the search for new early screening methods for ovarian cancer was found and a new ovarian cancer treatment is the focus of the study. In this study, real time quantitative PCR method to detect ovarian cancer and ovarian CXCL1 in benign lesions and normal tissues, and to further reveal its expression and clinical data of ovarian cancer between. Then constructed lentiviral CXCL1 gene expression vector, and through a series of in vitro experiments, CXCL1 in the process of growth and proliferation of ovarian cancer and its mechanism were investigated.Chapter 2 The study on the relationship between the expression of CXCUmRNA and the Clinical-pathology factor in ovarian tumorObjective:To investigate the relationship between the expression of CXCL1 (Chemokine (CXC motif) ligand 1 (melanoma growth stimulating activity, alpha) gene mRNA in ovarian tumor and the clinical-pathology factor.Methods:Trizol one-step method was used to extract the total RNA in ovarian tissue, and real-time fluorescence quantitative PCR technique was used to detect the expression copy number of CXCL1 gene in 80 cases of ovarian cancer,40 cases of ovarian benign lesions and 20 normal control. As well as the relationship between the expression of CXCL1 gene mRNA in ovarian tumor and the clinical-pathology factor was analyzed.Results:The expression copy number of CXCL1 mRNA in ovarian malignancy was higher than that in benign lesions as well as normal control. There was a difference significantly (P<0.01); In advanced ovarian cancer (Ⅲ-Ⅳstage), The expression of CXCL1 mRNA was also significantly higher than that in the patients with early stage (Ⅰ～Ⅱstage)(P=0.005). The results of Kaplan-Meier survival curve showed that there was no relationship between the expression of CXCL1 mRNA and the prognosis in ovarian cancer, and the analysis of Cox proportional hazard model also showed CXCL1 expression was not independent prognostic factors. For patients with ovarian cancer.Conclusions:The expression copy number of CXCL1 mRNA in ovarian malignant was higher significantly than that in benign lesions as well as normal control. It was suggesting that the over-expression of CXCL1mRNA should be related with occurrence and development of ovarian malignantChapter 3 The construction and identification of CXCL1 gene expression vectorObjective:Using the second-generation lentiviral vector system to constructe lentiviral CXCL1 gene expression vector, and exploring the methods of gene transfer more efficient and reliable than traditional methods, laying a good basis for gene targeted therapy in the further.Methods:Amplification CXCL1 full sequence of its connection with the lentiviral vector pWPI sequenced by BLAST search, identification of lentiviral expression vectors. The recombinant lentiviral vector transfected 293T cells, identified by fluorescence microscopy after 48 hours, lentivirus infection and packaging success.Results:The recombinant lentiviral vector CXCL1-pWPI sequencing by BLAST, and the CXCL1 gene homology 99%.Conclusion:Successfully constructed recombinant lentivirus CXCL1 expression system. Laying the foundation for CXCL1 gene function in future research.Chapter 4 Experimental study of CXCL1-mediated gene on the biological function of epithelial ovarian cancer in vitroObjective:To understand the function of CXCL1 impact on ovarian cancer,analysis CXCL1 in ovarian cancer growth and metastasis the role of the process.Methods:The successful extraction of CXCL1 gene containing lentiviral particles, will be transfected into non-CXCL1 gene expression in ovarian cancer cell line A2780, by fluorescence microscopy and RT-PCR to confirm identification of stable transfected A2780 cell lines. And then cell growth, cell cycle experiment.Results:Lentiviral infection, we obtain stable expression of CXCL1 gene ovarian cancer cell line. MTT assay showed that, CXCL1 and control groups of growth curves were significantly different, CXCL1 group significantly faster cell growth. CXCL1 group of colony formation rate than the control group and the control group, the difference was statistically significant.Conclusion:The results show that cell function, CXCL1 can enhance the A2780 ovarian cancer cell growth and proliferation.