2.Effect of miR-30a-5p on the proliferation, apoptosis, invasion and migration of SMCC-7721 human hepatocellular carcinoma cells.
Hang DAI ; Bing KANG ; Deyu ZUO ; Guoqing ZUO
Chinese Journal of Hepatology 2014;22(12):915-920
OBJECTIVETo explore the effect of microRNA-30a-5p (miRNA-30a-5p) on the biological behavior of human hepatoma cells.
METHODSThe liver cancer cell line SMCC-7721 cells and the normal liver cell line L02 cells (control) were transiently transfected with miRNA-30a-5p mimics and an miRNA-30a-5p inhibitor by Lipofectamine 2000 (Life Technologies). miR-30a-5p mRNA expression was detected by quantitative real-time (q)PCR. Cell proliferation was evaluated using the Cell Counting Kit-8 (CCK-8) assay and apoptosis was assessed by flow cytometry.Invasion and migration were measured by transwell chamber assays. The SMCC-7721 cells was injected subcutaneously into nude mice to establish a tumor animal model.
RESULTSThe SMCC-7721 cells transfected with miRNA-30a-5p mimics showed significantly higher miRNA-30a-5p mRNA expression than the non-transfected SMCC-7721 cells and the transfected control L02 cells (P<0.01). The miRNA-30a-5p mRNA expression was significantly lower in the SMCC-7721 cells transfected with the miRNA-30a-5p inhibitor than the non-transfected SMCC-7721 cells the control L02 cells (P<0.01). The overexpression of miRNA-30a-5p inhibited the viability, colony formation rate, and invasion and migration abilities, as shown in the cells transfected with the miRNA-30a-5p mimics (P<0.05); in addition, the miRNA-30a-5p promoted proliferation of cells (P<0.05), as shown by more S phase cells detected by flow cytometry. SMCC-7122 cells transfected with miRNA-30a-5p mimics produced tumors with significantly higher average weight than tumors produced by SMCC-7122 cells that were untransfected or transfected with empty vector (both P<0.01).
CONCLUSIONOverexpression ofmiR-30a-5p had an inhibitory effect on cell proliferation, induced apoptosis, increased the number of cells in S phase, and markedly inhibited invasion and migration of SMCC-7721 HCC cells in vitro and in vivo.
Animals ; Apoptosis ; Carcinoma, Hepatocellular ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; Mice ; Mice, Nude ; MicroRNAs ; Neoplasm Invasiveness ; Transfection