The Role Of KIAA1199 Gene In The Development And Progression Of Ovarian Cancer

Background & Objective: Due to the improvement of oncology treatment has improved since the mid-1970 s,the mortality rate of ovarian cancer has decreased by more than 30 percent.However,ovarian cancer still has a high mortality rate.The reasons are as follows: 1.the more malignant epithelial carcinoma predominates in ovarian cancer;2.the ovaries are deep in the pelvic cavity and the onset of ovarian related diseases is subtle;3.today,there are no effective screening methods for ovarian cancer and definitive diagnosis at early stage is quite complicated.As a result,most of these diseases are already at an advanced stage when detected.Therefore,the present study wanted to clarify the relationship between KIAA1199 and the invasion and metastasis of ovarian cancer cells.And,we expected to understand the relevant pathways involved in the regulation of ovarian cancer by KIAA1199.This study will provide more options for prognosis determination and potential future therapeutic targets for ovarian cancer in the future.Methods:(1)Immunohistochemistry(IHC)was used to analyze the content of KIAA1199 in epithelial ovarian tumors,borderline ovarian tumors and normal ovarian tissues.To investigate the relationship between KIAA1199 and patients’ age,FIGO stage,cancer pathological type,lymphatic invasion,peritoneal dissemination,liver metastasis,and 5-year survival rate.(2)We guess that different ovarian cancer cell lines have different KIAA1199 protein levels.Then,the expression levels of KIAA1199 protein between four different ovarian cancer cell lines(A2780,HO-8910,CAOV-3 and OVCAR-3)were detected by western blot.(3)The sh RNA lentivirus was successfully transported into A2780 cells to silence the expression of KIAA1199.These A2780 cells were divided into experimental group(the expression of KIAA1199 silenced)and control group.In vitro,we performed Transwell assays to evaluate the migration ability of A2780 cells with knocking down KIAA1199(4)Gene expression profiling detection and functional enrichment analysis of KIAA1199 knockout A2780 cells and A2780 cells with negative transfection.(5)Through ALGGEN-PROMO website,we found STAT3 is potential transcription factor of KIAA1199 gene.Then,HO8910 cell chromatin fragment DNA was immunoprecipitated by STAT3 antibody,and KIAA1199 m RNA expression was detected by Chromatin Immunoprecipitation assay(Ch IP).(6)q RT-PCR and WB assays were performed to detect the expression of KIAA1199,p-STAT3 and STAT3 in HO8910 cells after treating with IL-6/STAT3 inhibitor and activator.Results:(1)The expression of KIAA1199 protein in ovarian cancer was significantly higher than that in non-cancer tissues.The expression of KIAA1199 protein was closely related to FIGO stage,peritoneal dissemination,liver metastasis and 5-year survival rate.However,it was not related to patient’s age,pathological type of cancer and lymph node metastasis.(2)The expression level of KIAA1199 protein was different among different ovarian cancer cell lines.(3)The results of transwell assays showed that the ability of invasion and migration was significantly inhibited when KIAA1199 expression was silenced in A2780 cells.(4)By comparing the expression profile data of the gene knockout experimental group and the control group,we screened 707 differentially expressed protein molecules,including 237 upregulated genes and 470 downregulated genes.Functional enrichment analysis suggested that the alteration of KIAA1199 gene was related to biological behaviors such as cell adhesion.(5)Ch IP assay detected that STAT3 could bind to the KIAA1199 promoter and this effect can be enhanced under the action of IL-6.(6)The expression levels of KIAA1199,P-STAT3 and STAT3 were significantly decreased or increased after treatment with cryptotanshinone or IL-6.Conclusion:(1)The expression of KIAA1199 is increased in ovarian cancer,and the overall survival rate of EOC patients with high KIAA1199 expression is lower.(2)In vitro,KIAA1199 can promote the migration and invasion of ovarian cancer cells and play a carcinogenic role in the development of ovarian cancer.(3)STAT3 had a transcriptional regulatory effect on KIAA1199 expression and the regulation was enhanced in the presence of IL-6(4)The expression of KIAA1199 protein in ovarian cancer cells was affected by stimulation or inhibition of IL6/STAT3 pathway,suggesting that the IL6/STAT3 pathway can influence the involvement of KIAA1199 in the migration and invasion of ovarian cancer cells.