Mechanism of miR-30c over-expression inhibiting malignant phenotypes of cervical cancer cells / 中国病理生理杂志

ABSTRACT

AIM:To investigate the molecule mechanism of microRNA(miR)-30c over-expression inhibiting malignant phenotypes of cervical cancer cells.METHODS:Cervical cancer cell lines C33A,HeLa,SiHa and CaSki were transfected with pGenesil-1-miR-30c plasmid using Lipofectamine 2000 kit,and the expression of miR-30c was determined by TaqMan real-time PCR.The cell viability inhibition rate,colony formation ability,migration rate and apoptotic rate were measured by MTT assay,colony formation assay,Transwell experiment,and flow cytometry with Annexin V-FITC staining. The protein expression of Bax,Bcl-2, matrix metalloproteinase(MMP)-9, MMP-13 and tissue inhibitor of metalloprotei-nase-1(TIMP-1)was detected by Western blot.RESULTS:The expression levels of miRNA-30c in the cervical cancer cell lines transfected with pGenesil-1-miR-30c plasmid were significantly higher than those in negative control groups(cell lines transfected with pGenesil-1 plasmid)(P<0.01).Significantly increased cell viability inhibition rate,and decreased colony formation ability and migration rate were found in the cervical cancer cell lines over -expressing miR-30c as compared with negative control groups(P<0.05).The apoptotic rate in the cervical cancer cell lines over-expressing miR-30c was dramatically higher than that in control groups(P<0.05).Over-expression of miR-30c in cervical cancer cells promoted the protein expression of Bax and TIMP-1,and decreased the protein expression of Bcl-2 and MMP-13(P<0.05 or P<0.01).CONCLUSION:Over-expression of miR-30c significantly inhibits the viability and migration,and induces apop-tosis of cervical cancer cells.The mechanism may be related to activating apoptosis pathway and inhibiting MMP-13 protein expression.