DAMPs对人肝癌HepG2细胞增殖能力影响的机制研究

Mechanism of the Effect of DAMPs on Human Hepatocellular Carcinoma HepG2 Cells

目的:探讨在损伤相关模式分子(DAMPs)诱导下人肝癌HepG2细胞增殖能力变化的发生机制。方法:将HepG2细胞分为对照组,DAMPs组,IL-6 siRNA组,IL-6 siRNA+DAMPs组;MTT比色法检测HepG2细胞增殖能力的变化;实时荧光定量PCR法检测IL-6及STAT3 mRNA的表达;Western blot法测定HepG2细胞STAT3蛋白和磷酸化STAT3蛋白的表达。结果:DAMPs组细胞增殖能力较对照组显著增强,IL-6 mRNA及STAT3 mRNA的表达较对照组显著上调,差异均有统计学意义(P<0.01);Western bot法检测出四组STAT3蛋白表达无明显差异,而DAMPs组磷酸化STAT3蛋白的表达高于对照组,差异有统计学意义(P<0.01)。结论:DAMPs可能通过IL-6/STAT3信号通路促进人肝癌HepG2细胞的增殖。

Objective： To investigate the mechanism of human hepatocellular carcinoma HepG2 cells pro- liferation induced by DAMPs. Methods： HepG2 cells were divided into four groups： control group, DAMPs group, IL-6 siRNA group, and IL-6 siRNA＋DAMPs group. MTT assay was applied to investigate the proliferation of HepG2 cells. Expression of IL-6 and STAT3 mRNA in HepG2 cells was measured by quantitative RT-PCR, and the protein expression level of STAT3 and p-STAT3 was detected by Western blotting. Results： MTT assay demonstrated that the pro- liferation of HepG2 cells was enhanced when exposed to DAMPs （P〈0.01, vs. control）. The result of quantitative RT-PCR showed that the expression of IL-6 mRNA and STAT3 mRNA in- creased markedly when exposed to DAMPs（P〈0.01, vs. control）. The protein expression level of STAT3 among the four groups have no differences, but the p-STAT3 in DAMPs group werehigher than in control group （P〈0.01）. Conclusion： DAMPs may promote the proliferation of HepG2 cell by IL-6/STAT3 signaling pathway.