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Research Article Open Access
Background: To investigate the effect of alpha fetoprotein (AFP) on benzyl-isothiocyanate (BITC) arresting cell cycle in human liver cancer cells line in vitro, and explore the possible role mechanism of BITC inhibited proliferation of hepatocellular carcinoma (HCC) cells. Methods: In this study, we selected HCC cells lines, Bel 7402 and HLE for test. Fluorescent microscopy and Western blotting were applied to observe the transfected effect and expression of proteins; MTT assay the proliferation of HCC cells; Flow cytometry method was used to detect the cellular cycle; RNA interference was used to test and expressed vector constructed technology were performed to silence and induce expression of AFP, respectively. Results: Analysis showed BITC had a significant inhibitory effect on the proliferation of HCC cells in dosedependent manner, inhibitory effect was enhanced while Bel 7402 cells were transfected with AFP-siRNA vectors but attenuated in HLE cells while transfected with pcDNA3.1-afp vectors. The growth ratio of AFP-siRNA transfected Bel 7402 cells and pcDNA3.1-afp transfected HLE cells followed treated with 80 μmol/L BITC were (42.43 ± 4.92)% (P<0.05 vs Bel 7402 cells group) and (40.13 ± 4.99)% (P<0.05 vs HLE cells group). BITC could obviously induce cell cycle G2/M phase arrest in theses HCC cells; The induction effect was enhanced in AFP-siRNA transfected Bel 7402 cells but attenuated in pcDNA3.1-afp transfected HLE cells. BITC inhibited the expression of cell cycle related proteins, cyclin B1, CDK1, Cdc25c but stimulated expression of Weel in Bel 7402 cells and HLE cells, and such effect was enhanced in AFP-siRNA transfected Bel 7402 cells but attenuated in pcDNA3.1-afp transfected HLE cells. Conclusions: BITC could inhibit the growth of HCC cells and induce cell cycle G2/M phase arrest through down regulating the expression of cyclin B1, CDK1, Cdc25c and up regulating the expression of Weel; AFP played an antagonistic role in BITC arresting cell cycle in HCC cells.
Alpha-fetoprotein (AFP), Benzyl isothiocyanate (BITC), Hepatocellular carcinoma cells, Cell cycle arrest, Alpha-fetoprotein (AFP), Benzyl isothiocyanate (BITC), Hepatocellular carcinoma cells, Cell cycle arrest