Primary Research Of MiRNA-497 Suppresssing Ovarian Cancer Cell Proliferation Through Regulating FASN Expression

[Objective]To test the expression of miRNA-497 in ovarian cancer tissues/cells and the effect of miRNA-497 on ovarian cancer cell proliferation, explore the relationship between miRNA-497 and FASN, produce a new therapeutic direction for clinical diagnosis and treatment of ovarian cancer, provide a new experimental study based on selective targeting drug use.[Methods](A) The ovarian cancer specimens and normal ovarian tissues were collected in zhongda Hospital of Southeast University, which were confirmed by pathology and the patients without preoperative chemotherapy. qPCR was used for testing the expression of miRNA-497 and FASN in 10 cases of epithelial ovariancancer tissues and 10 cases of normal epithelial ovarian tissues,person correlation analysis method can reveal the correlation between miRNA-497 and FASN.(B) The gene tran suription expression of miRNA-497 was detected by qPCR in the human ovarian cancer SKOV3 and A2780.(C) Cell transfection was afford to SKOV3 ovarian cancer cell line. miRNA-497 mimics and miRNA-497 inhibitor were considered as experimental groups,miRNA-497 mimics control and miRNA-497 inhibitor control were taken as negative control.and black transfection was regarded as blank control. After that CCK8 experiment was used to test changes of different group cell’s proliferation.(D) FASN mRNA expression of different groups was tested by PCR in different groups after transfection.(E) The changes of FASN protein expression in different group after transfection was analysised by Western blot.[Results](A) The expression of miRNA-497 in epithelial ovarian cancer tissues was significantly lower than normal ovarian tissue, but the expression of FASN gene was abnormally higher;Pearson correlation analysis showed that the expression of miRNA-497 and FASN mRNA in ovarian cancer tissue and normal ovarian tissues was negative correlation (r=0.45884, p< 0.01).(B)In the human ovarian cancer cell SKOV3 and A2780,the expression of miRNA-497 was high in the SKOV3 than those in A2780 and the SKOV3 was chosen as the target cells.(C) miRNA - 497 can inhibit the proliferation of ovarian cancer cell lines SKOV3.CCK8 results indicated that when miRNA-497 mimics was transfected into SKOV3 cells,the ability of cell proliferation was significantly suppressed compared with the control group (p<0.05). But in miRNA-497 inhibitor’s group, the ability of cell proliferation was obviously improved,which was compared with the control group (p<0.05). The result showed that miRNA-497 can inhibit the proliferation of ovarian cancer cell lines SKOV3 (p<0.05), but the function will not increasewith time (p> 0.05).(D) miRNA-497 can inhibit the expression of FASN. In the miRNA-497 mimics group, FASN mRNA transcription and protein expression were both significantly reduced compared with the control group (p<0.05),however miRNA-497 inhibitor increased the expression of FASN mRNA and protein. This suggest that miRNA-497 can inhibit the expression of FASN in ovarian cancer.[Conclusions](A) The expression of miRNA-497 in epithelial ovarian cancer tissues was significantly lower than normal ovarian tissue, but the expression of FASN gene was abnormally higher,and the expression of miR-497 and FASN have negative correlation.(B)miRNA-497 significantly inhibited the proliferation of ovarian cancer SKOV3 cells in vitro, had effects on the biological characteristics of ovarian cancer cells, but the effect had no obvious correlation with time..(C) miRNA-497 can regulate the expression of FASN, FASN is the target gene of miRNA-497.miRNA-497 is probably affect ovarian cancer cell proliferation by inhibiting the expression of FASN,but the related mechanism remains unknown.The result provides experimental basis for the further research of function of miRNA-497 in ovarian cancer and a new angle for selectively targeted therapy of ovarian cancer.