Effects of microRNA-100 on mitosis block and Polo-like kinase 1 expression in hepatocellular carcinoma cells

doi:10.3969/j.issn.0529-1356.2014.01.013

Abstract

Abstract:

Objective To investigate the effect of microRNA-100(miR-100) on mitosis block and Polo-likekinase 1(PLK1) expression in hepatocellular carcinoma cells. Methods Expression of miR-100 and PLK1 was determined by quantitative real-time PCR (qRT-PCR) and immunofluorescence (IF) in hepatocellular carcinoma HepG2 cells. HepG2 cells were transiently transfected by miR-100 mimic with Cy3 fluorescence labeled through Oligofectamine liposomes, followed by cell mitosis and PLK1 expression analysis. Results Expression level of miR-100 decreased whereas PLK1 mRNA and protein expression abnormally increased in HepG2 cells compared with the normal hepatocytes. At 48hours after transfection of miR-100 mimic, the mitotic index of HepG2 cells in experimental group was significantly less than that of control cells. Western blotting analysis indicated that PLK1 protein expression levels were significantly reduced in experimental cells compared with the control cells. Immunocytochemical results showed that PLK1 protein in the nucleus almost disappeared in meta/ana-phase and telophase of mitosis. Conclusion MiR-100 may inhibit the expression of PLK1 protein, which leads to mitotic block of hepatocellular carcinoma cells.

Key words: Hepatocellular carcinoma, Mitosis, MicroRNA-100, Polo-like kinase 1, Real-time quantitative PCR, Western blotting, Immunofluorescence

FAN Bing-lin JI Xiao-hui ZHU Wu-ling*. Effects of microRNA-100 on mitosis block and Polo-like kinase 1 expression in hepatocellular carcinoma cells[J]. AAS, 2014, 45(1): 70-73.

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