Effects Of The TCP1 On Proliferation And Invasion Ability Of Ovarian Cancer Cells And Prognosis

Objective: To investigate the expression of TCP1 in epithelial ovarian carcinoma(EOC)and its correlation with prognosis and clinicopathological parameters of the patients.To observe the effect of TCP1 gene expression difference on malignant behavior of tumor cells at the cellular level,and to explore the potential mechanism of TCP1 involvement in and regulation of malignant behavior of ovarian cancer cells.Methods:1.Tissue specimens of the normal ovary,ovarian cystadenoma,borderline ovarian cancer(OC)and EOC confirmed by pathology were collected to detect the expression of TCP1 protein in these four different ovarian tissues by immunohistochemical method(IHC).2.The expression level of TCP1(IOD/area)in each tissue section was calculated by Image-Pro Plus software.The expression difference of TCP1 in ovarian tissues of different histological types was analyzed by SPSS 20.0 software,and the relationship between the expression level of TCP1 and clinicopathological parameters such as FIGO stage and tumor grade in patients with EOC was analyzed.3.In Kaplan-Meier plotter online database,the TCP1 probe(Affymetrix ID: 222010＿AT)was used to analyze the correlation between TCP1 expression and overall survival(OS)and tumor-free survival(PFS)in EOC.4.The expression of TCP1 protein in IOSE-80(normal ovarian epithelial cells)SKOV3and A2780(EOC cell lines)was detected by Western blot.5.shTCP1 lentivirus was used to infect the A2780 cells with high expression of TCP1,and stable TCP1 knockdown cell line A2780/TCP1-KD was constructed,while negative control lentivirus was used to construct negative control cell line A2780/TCP1-NC.6.The proliferation,migration,and invasion ability of A2780/TCP1-NC and A2780/TCP1-KD cells were detected by the MTT method and Transwell method.7.The protein expression of key regulatory factors in PI3K/Akt/mTOR pathway of A2780/TCP1-NC and A2780/TCP1-KD cells was detected by Western Blot.Results:1.The quantitative analysis of TCP1 expression in different ovarian tissue types showed that the expression of TCP1 in borderline ovarian cancer and EOC was higher than that in normal ovarian tissue and ovarian cystadenoma tissue,and the difference was statistically significant.2.The expression of TCP1 in EOC tissues was correlated with tumor grade and FIGO stage.Different tumor grades and FIGO stages showed statistically significant differences in the expression of TCP1.3.Through Kaplan-Meier plotter online database,it was found that the expression of TCP1 was negatively correlated with the prognosis of OC patients.The higher the expression of TCP1,the worse the prognosis.4.Western Blot results showed that the expression of TCP1 in EOC cell A2780 was higher than that in normal ovarian cell IOSE-80.5.Western Blot verification: A2780 cells were infected with lentivirus,and the stable transgenic cell line A2780/TCP1-KD cells and negative control cell line A2780/TCP1-NC were successfully constructed.6.Compared with A2780/TCP1-NC cells,the proliferation,migration,and invasion ability of A2780/TCP1-KD cells was inhibited.7.Western Blot results showed that TCP1 knockdown could down-regulate the expression of p-Akt and p-mTOR in PI3K/Akt/mTOR pathway in A2780 cells.Conclusion:1.TCP1 protein level is highly expressed in EOC,and its expression level is negatively correlated with the prognosis of patients.Therefore,TCP1 may become a new indicator to evaluate the survival and prognosis of OC patients.2.The down-regulated expression of TCP1 in OC cells inhibits the proliferation,migration,and invasion ability of cells.The potential mechanism may be that the knockdown of TCP1 affects the malignant behavior of OC cells by inhibiting the PI3K/Akt /mTOR signaling pathway,so TCP1 may become a new target for the treatment of OC.