MicroRNA-100 Is Downregulated In Cervical Cancer And Inhibits Cell Proliferation,Migration And Invasion By Targeting Shp2

Objective: To investigate the expression of miRNA-100 in cervical cancer and whether changing its expression level can affect the malignant biological behavior of cervical cancer cells.Methods:1.qRT-PCR was used to detect the expression of miRNA-100 in cell lines(cervical cancer cells HeLa,SiHa,and human immortalized keratinocytes HaCaT)and 21 untreated samples of cervical cancer(cancerous and adjacent tissues).2.Western blotting was used to detect the expression of Shp2 protein in cervical cancerous and adjacent tissues.3.Cervical cancer cell lines were infected with lentiviral vectors to establish Hela cell lines that knocked down miRNA-100 expression and SiHa cell lines that over-expressed miRNA-100 expression.4.Studying the biological functions of miRNA-100 using constructed stable transfected cell lines.(1)Ki67 protein positive rates in HeLa and SiHa cells(Ki67 protein expression level can reflect the malignant proliferation ability of tumor cells)were detected by flow cytometry.(2)Colony forming ability of HeLa and SiHa cells were detected by plate cloning test.(3)Migration and invasion of HeLa and SiHa cells were detected by transwell test.5.Western blotting was used to detect changes in Shp2 protein expression and p-ERK and p-AKT expression in downstream signaling pathways.Results:1.Compared with human immortalized keratinocytes HaCaT,the expression of miRNA-100 was down-regulated in HeLa and SiHa;compared with adjacent tissues,miRNA-100 was down-regulated in cancerous tissues.2.The expression level of Shp2 protein in cancerous tissues is higher than that in adjacent tissues.3.Successfully constructed HeLa cell lines that knocked down miRNA-100 expression and SiHa cell lines that over-expressed miRNA-100 expression.4.Effects of miRNA-100 on the malignant biological behavior of cervical cancer.(1)Effect of miRNA-100 on the malignant proliferation ability of cervical cancer cells: flow cytometry experiments showed that after knocking down miRNA-100 expression in HeLa cells,the Ki67 protein positive rate of the cells increased;after miRNA-100 was over expressed in SiHa cells,the Ki67 protein positive rate of the cells decreased.Experiments have shown that miRNA-100 can inhibit the malignant proliferation of cervical cancer cells.(2)Effect of miRNA-100 on the ability of cervical cancer cells to form clones:plate cloning experiments have shown that after knocking down miRNA-100 expression in HeLa cells,the colony formation rate of the cells is significantly increased;after miRNA-100 is over expressed in SiHa cells,the colony formation rate of the cells was significantly reduced.Experiments have shown that miRNA-100 can inhibit the ability of cervical cancer cells to form clones.(3)Effect of miRNA-100 on cervical cancer cell migration and invasion ability:transwell experiments show that after miRNA-100 expression is knocked down in HeLa cells,the cell migration and invasion ability is significantly enhanced;after miRNA-100 is over expressed in SiHa cells,the ability of cells to migrate and invade was significantly reduced.Experiments have shown that miRNA-100 can inhibit the migration and invasion of cervical cancer cells.5.Effect of miRNA-100 on the expression of Shp2 protein.After knocking down the expression of miRNA-100 in HeLa cells,the expression of Shp2 protein increased,as well as the expressions of downstream p-ERK and p-AKT.However,when miRNA-100 was over expressed in SiHa cells,the expression of Shp2 protein decreased,as well as the expressions of downstream p-ERK and p-AKT.The study shown that miRNA-100 can inhibit the the expression of Shp2 protein and downstream p-ERK,p-AKT.Conclusion: miRNA-100 is underexpressed in cervical cancer tissues and cell lines.Up-regulating miRNA-100 expression can inhibit malignant biological behavior of tumor cells,while down-regulating miRNA-100 expression can promote malignant biological behavior of tumor cells.The biological role of miRNA-100 in cervical cancer may be achieved in part by regulating the expression of Shp2 protein.