Alpha Fetoprotein Plays Antagonistic Role in Benzyl-Isothiocyanate ArrestingCell Cycle in Liver Cancer Cells
Alpha Fetoprotein Plays Antagonistic Role in Benzyl-Isothiocyanate ArrestingCell Cycle in Liver Cancer Cells
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DOI：
10.4172/2167-7700.1000195
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发表时间：
2016-03
期刊：
影响因子：
3.3
通讯作者：
Mingyue Zhu;Wei Li;Xu Dong;P. Zhou;Junli Guo;Mengsen Li
中科院分区：
文献类型：
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作者：
Mingyue Zhu;Wei Li;Xu Dong;P. Zhou;Junli Guo;Mengsen Li
背景：观察甲胎蛋白(AFP)对异硫氰酸苄酯(BITC)体外阻断人肝癌细胞周期的影响，探讨BITC抑制肝癌细胞增殖的可能作用机制。方法：本研究以人肝癌细胞系Bel 7402和HLE为研究对象。用荧光显微镜和Western blotting观察细胞的转染率和蛋白质的表达，用四甲基偶氮唑盐比色法检测肝癌细胞的增殖，用流式细胞仪检测细胞周期，用RNA干扰检测和构建表达载体分别沉默和诱导甲胎蛋白的表达。结果：BITC对肝癌细胞的增殖有明显的抑制作用，且呈剂量依赖关系，AFP-siRNA载体对Bel 7402细胞的抑制作用增强，而对HLE细胞的抑制作用减弱。80μ的BITC处理后，转导AFP-siRNA的Bel7402细胞和转导AFP的HLE细胞的生长率分别为(42.43±4.92)%和(40.13±4.99)%(P<；005vs HLE细胞组)。BITC能明显诱导肝癌细胞周期G2/M期停滞，AFP-siRNA对Bel 7402细胞的诱导作用增强，而对HLE细胞的诱导作用减弱。BITC抑制Bel 7402和HLE细胞周期相关蛋白Cyclin B1、CDK1、CDc25c的表达，但促进Weel的表达，AFP-siRNA对Bel 7402细胞的作用增强，而对HLE细胞的作用减弱。结论：BITC通过下调Cyclin B1、CDK1、CDC25c的表达，上调Weel的表达，抑制肝癌细胞生长，诱导细胞周期停滞于G2/M期，AFP对BITC阻断肝癌细胞周期具有拮抗作用。
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.