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目的:探讨转化生长因子β1(transforming growth factorβ1,TGF-β1)体外对肝癌细胞干性特征的影响。方法:以转化H-ras的鼠肝肿瘤细胞LPC-H-ras和人肝癌细胞株PLC/PRF/5为研究对象,利用MTT法和多细胞球体培养法检测TGF-β1对肿瘤起始细胞增殖的抑制作用,FCM法检测细胞表面分子CD133的表达以及活性氧(reactive oxygen species,ROS)的水平;实时荧光定量PCR(real-time fluorogentic quantitative-PCR,RFQ-PCR)检测干细胞样基因的表达水平。结果:TGF-β1在体外能够改变LPC-H-ras细胞的形态并抑制细胞的生长;与未处理组相比,TGF-β1作用后,LPC-H-ras细胞表面CD133的表达降低,多细胞球体形成能力减弱,干细胞样基因的表达受到抑制;同样,在PLC/PRF/5细胞中,TGF-β1也能抑制CD133的表达、AFP基因表达以及多细胞球体的形成;另外,在LPC-H-ras细胞中,TGF-β1可诱导ROS表达升高,ROS的抑制剂N-乙酰半胱氨酸具有削弱TGF-β1诱导CD133表达下调的作用。结论:TGF-β1能够抑制肝癌起始细胞的增殖及干性,其作用机制可能与ROS表达上调有关。
Objective: To investigate the effect of transforming growth factor β1 (TGF-β1) on the hepatocellular carcinoma cells in vitro. Methods: LPC-H-ras and PLC / PRF / 5 cells were transformed into H-ras, and MTT assay and multicellular spheroid culture were used to detect the proliferation of tumor-initiating cells (FCM) was used to detect the expression of cell surface molecule CD133 and the level of reactive oxygen species (ROS). The expression of stem cell-like genes was detected by real-time fluorogentic quantitative-PCR (RFQ-PCR) . Results: TGF-β1 could change the morphology and inhibit the growth of LPC-H-ras cells in vitro. Compared with the untreated group, the expression of CD133 on the surface of LPC-H-ras cells decreased after TGF-β1 treatment. Similarly, in PLC / PRF / 5 cells, TGF-β1 also inhibited the expression of CD133, AFP gene expression and the formation of multicellular spheroids. In addition, in the LPC-H -ras cells, TGF-β1 can induce ROS expression increased, N-acetyl cysteine, an inhibitor of ROS, has the effect of down-regulating the expression of CD133 induced by TGF-β1. Conclusion: TGF-β1 can inhibit the proliferation and stem cell proliferation of hepatocellular carcinoma cells. Its mechanism may be related to the up-regulation of ROS expression.