TGF-β1通过cyclinD3对肝细胞癌增殖能力的双重调控作用机制

The double regulation mechanisms of TGF-β1 in the proliferation of hepatocellular carcinoma by cyclin D3

目的探讨肝细胞癌细胞分泌的转化生长因子β1(transforming growth factor-β1,TGF-β1)对癌细胞增殖能力的调控机制。方法免疫荧光检测TGF-β1的表达能力。构建TGF-β1下调肝细胞癌细胞株。克隆形成实验及流式细胞术检测不同干预下Hepa1-6细胞的成瘤及细胞周期的变化。Western blotting检测cyclin D3的变化。结果免疫荧光检测显示多株肝细胞癌细胞均具有表达TGF-β1的能力。下调TGF-β1后Hepa 1-6细胞的克隆形成能力减弱,而阻断TGF-β1信号后其克隆形成能力增强。流式检测结果表明下调TGF-β1后细胞出现G1期阻滞,而加入LY-364947后结果则相反。Western blotting检测结果显示TGF-β1对cyclin D3具有双重调控作用。结论 TGF-β1有可能通过影响cyclin D3从而达到对肝细胞癌细胞增殖能力的双重调控作用。

Objective To investigate the regulation mechanisms of transforming growth factor-β1 （TGF-β1） in the proliferation of hepatocellular carcinoma. Methods Immunofluorescence was used to check the expression of TGF-β1. Colony formation and flow-cytometry experiments were used to detect the proliferation of Hepa 1-6 cells in different con- ditions. And the expression of cyclin D3 was tested by Western blotting. Results Immunofluorescence showed that hepatocellular carcinoma cell line had the ability to express TGF-131. The ability to clone the Hepa 1-6 cells was de- creased after knock-down TGF-β1, and the ability of cells to clone the Hepa 1-6 cells was increased after blocking TGF- β1, Knock-down TGF-β1 impaired the colony formation and induced G1 arrest, but adding LY-364947 had an opposite. Western blotting result indicated that TGF-β1 played double roles in the regulation of cyelin D3. Conclusion TGF-β1 may control the proliferation of hepatocellular carcinoma by regulating cyclin D3.