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目的:探讨白藜芦醇(RES)抑制白血病K562细胞增殖的机制及非折叠蛋白应答(UPR)对其机制的影响。方法:选取不同浓度的RES干预K562细胞,并且用RES(100μmol/L)干预K562细胞不同时段;利用MTT法检测细胞存活率;利用流式细胞仪分析细胞周期并检测细胞凋亡率;利用蛋白质印迹法检测凋亡早期蛋白及UPR靶点蛋白表达水平;利用实时定量PCR法检测GRP78和CHOP mRNA的表达。结果:RES导致K562细胞细胞存活率明显减少,并呈剂量依赖方式;RES的生长抑制作用主要是阻断细胞周期中的G1期,G2/M期的变化较小;RES处理的K562细胞没有显著的细胞凋亡。RES可提高GRP78mRNA的表达水平,呈剂量及时效依赖方式;RES对CHOP mRNA水平也有诱导作用,但只有在RES浓度较大时(50μmol/L)或作用时间较长时(8h)才开始上调,并且上调幅度较小。蛋白质印迹分析证实RES提高UPR靶点(GRP78、GRP94、磷酸化eIF2a和剪接作用XBP-1)的蛋白表达水平。结论:RES优先诱导UPR存活分支,表明UPR参与RES抑制白血病K562细胞增殖的机制。
OBJECTIVE: To investigate the mechanism of resveratrol inhibiting the proliferation of leukemia K562 cells and the effect of non-folded protein response (UPR) on its mechanism. Methods: K562 cells were treated with different concentrations of RES, and K562 cells were treated with RES (100μmol / L) for different time periods. The cell viability was detected by MTT assay. Cell cycle was analyzed by flow cytometry and the apoptosis rate was detected. The expression of GRP78 and CHOP mRNA was detected by real-time PCR. Results: The survival rate of K562 cells induced by RES was significantly reduced and in a dose-dependent manner. The growth inhibition of RES mainly blocked the G1 phase in the cell cycle, and the changes in G2 / M phase were small. RES-treated K562 cells showed no significant difference Apoptosis. RES could increase the expression of GRP78 mRNA in a dose-and time-dependent manner. RES also induced the level of CHOP mRNA. However, RES increased only when RES concentration was high (50μmol / L) or longer (8h) And the rate of increase is small. Western blot analysis confirmed that RES increased the protein expression levels of UPR targets (GRP78, GRP94, phospho-eIF2a and splicing XBP-1). Conclusion: RES preferentially induces the survival of UPR branches, indicating that UPR is involved in the mechanism of RES inhibiting the proliferation of leukemia K562 cells.