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目的观察人喉表皮样癌(human laryngeal epidermoid,Hep-2)细胞自噬基因时空的表达及白藜芦醇的干预研究。方法在2014年1—12月用RT-PCR和Western blot分别分析不同剂量和不同时间白藜芦醇对自噬基因的m RNA和蛋白水平的影响。组间差异统计分析采用One-way ANOVA,P<0.05为差异有统计学意义。结果白藜芦醇剂量和时间依赖性地减少Hep-2细胞的存活率,以80μm作用的效果作为显著,白藜芦醇剂量和时间依赖性地促进Hep-2细胞自噬,以40μm和第6 h时最为显著,其能增加LC3Ⅱ与LC3Ⅰ的比例,促进beclin-1蛋白和m RNA的表达。结论白藜芦醇通过不同剂量和不同时间能依赖性地促进Hep-2细胞自噬,抑制自噬则白藜芦醇促Hep-2细胞凋亡作用减弱。
Objective To observe the spatiotemporal expression of autophagy gene in human laryngeal epidermoid carcinoma (Hep-2) cells and the intervention of resveratrol. Methods From January to December 2014, RT-PCR and Western blot were used to analyze the effects of resveratrol on the m RNA and protein levels of autophagy genes at different doses and times. Statistical analysis of differences between groups using One-way ANOVA, P <0.05 was considered statistically significant. Results Resveratrol dose-dependently and time-dependently decreased the survival rate of Hep-2 cells. The effect of 80μmol / L resveratrol was significant. Resveratrol promoted the autophagy in Hep-2 cells in a dose-dependent and time-dependent manner. 6 h when the most significant, which can increase the proportion of LC3 Ⅱ and LC3 Ⅰ, promote beclin-1 protein and m RNA expression. Conclusion Resveratrol can promote the autophagy of Hep-2 cells at different dosage and time-dependently, while the inhibition of autophagy decreases resveratrol-induced apoptosis of Hep-2 cells.