Effects Of MiR-200c-3p On The Proliferation, Migration And Invasion Of Renal Cancer Cells

OBJECTIVE: To detect the expression of microRNA-200c-3p(miR-200c-3p)in renal cancer cell lines and explore its effects on the proliferation,apoptosis,migration and invasion of renal cancer cell lines.To explore whether miR-200c-3p can affect the epithelial-mesenchymal transition process of renal cancer cells by regulating ZEB2(zinc finger E-box-binding homeobox2).Methods:(1)The expression of miR-200c-3p in human embryonic kidney cells 293 T,human adrenocortical small cell cancer cell SW3,human kidney cancer cell lines ACHN and786-O was detected by real-time quantitative PCR.(2)The experiment was divided into four groups: Blank group,miR-200c-3p mimics group,miR-200c-3p inhibitor group,and NC group.Transfect ACHN and 786-O cells respectively,and detect miR-200 c after cell transfection.-3p expression level.(3)The effect of miR-200c-3p on the proliferation of renal cancer cells was detected by MTT method,and the apoptosis rate of renal cancer cells was detected by flow cytometry.(4)Transwell test was used to detect the effect of miR-200c-3p on the invasion and migration ability of renal cancer cells.(5)Bioinformatics software was used to predict the target gene of miR-200c-3p as ZEB2,and Western Blot was used to detect the expression of ZEB2 protein and EMT pathway-related proteins.Results:(1)The expression levels of miR-200c-3p in human kidney cancer cell lines ACHN and 786-O were significantly reduced.Compared with human embryonic kidney cells 293 T,the differences were statistically significant(P <0.01,P <0.01).There was no significant difference in the expression level of miR-200c-3p in human embryonic kidney cells 293 T and human adrenocortical small cell cancer cell SW3(P> 0.05).This shows that miR-200c-3p is down-regulated in renal cancer cells,and miR-200c-3p may be involved as a tumor suppressor gene in the genesis and development of renal cancer.(2)Compared with NC group,miR-200c-3p mimics group had significantly higher expression levels of miR-200c-3p in ACHN and 786-O cells,and the difference was statistically significant(P <0.01),P <0.01).Compared with the NC group,the expression level of miR-200c-3p in the transfected miR-200c-3p inhibitor group was reduced,and the difference was statistically significant(P <0.01,P <0.01).Compared with the NC group and the Blank group,the expression of miR-200c-3p in the two cells was not statistically significant(P> 0.05,P> 0.05).(3)In ACHN and 786-O renal cancer cells,compared with the NC group of the control group,the miR-200c-3pmimics group showed growth inhibition at 24 hours after transfection,and the growth inhibition continued with time.Increased,the proliferation of miR-200c-3p mimics group was significantly inhibited after 72 hours,and the differences were statistically significant(P <0.05,P <0.05);miR-200c-3p inhibitor group was 24 hours after transfection The cell proliferation increased,and the cell proliferation significantly increased after 72 hours,and the differences were statistically significant(P <0.05,P <0.05);there was no statistically significant difference between the NC group and the Blank group(P> 0.05,P > 0.05).(4)The early apoptosis rate of miR-200c-3pmimics group in 786-O renal cancer cells was23.52%,which was significantly higher than that in Blank group(6.21%),miR-200c-3pinhibitor group(2.96%),and NC group.(9.78%);the late apoptosis rate of miR-200c-3pmimics group was 10.45%,which was higher than that of miR-200c-3pinhibitor group(4.2%)and NC group(7.81%).The same phenomenon also occurred in another type of kidney cancer cell ACHN.The early apoptosis rate and late apoptosis rate(15.21%,13.01%)in the miR-200c-3pmimics group were higher than those in the Blank group(2.36%,4.82%),miR-200c-3pinhibitor group(2.48%,3.34%),NC group(3.78%,3.61%).(5)Compared with the NC group in the control group,the number of cells passing through the Transwell compartment of renal cancer cells in the miR-200c-3pmimics group was significantly reduced,and the ability to invade and migrate was reduced(P <0.01,P <0.05,);miR-200 c In the-3pinhibitor group,the number of renal cancer cells passing through the Transwell compartment increased significantly,and the ability to invade and migrate was enhanced(P <0.05,P <0.05);there was no significant difference between the two in the Blank group(P>0.05).(6)Western Blot results showed that compared with NC group and Blank group,the expression level of ZEB2 in miR-200c-3p mimics group was significantly reduced(P<0.01),while the expression level of E-cadherin increased and the expression level of fibronectin decreased(P<0.01,P<0.01).Conclusion: 1.miR-200c-3p is lowly expressed in renal cancer cells ACHN,786-O,and miR-200c-3p may play a role as a tumor suppressor gene in renal cancer cells.2.miR-200c-3p plays a regulatory role in the biological function of renal cancer cells.Overexpression of miR-200c-3p can inhibit the proliferation of renal cancer cells,promote the apoptosis of renal cancer cells,and reduce the ability to invade and migrate.3.miR-200c-3p may affect the EMT process of renal cancer cells by regulating the expression of ZEB2.The conclusion of this study suggests that miR-200c-3p plays a regulatory role in the biological function of renal cancer cells,and may be one of the key miRNAs that affect the invasion,migration,and apoptosis of renal cancer cells.It is expected to become a diagnostic and diagnostic tool for renal cancer.Important targets in therapy.