大蒜素对重组质粒转染后Jurkat细胞周期和凋亡的影响

摘要

Objective:To study cell cycle and apoptosis on transfected-Jurkat cells induced by allicin,to discuss mechanism of action of allicin.Methods:The sequence of CXCR4 promoter gene was cloned,and was inserted into the reporter vector PgL4.17 to construct luciferase reporter vector PgL4.17-CXCR4,the recombinant plasmids were transfected into Jurkat cells.The lasting transfected cells were allocated into different dose groups,treated with the allicin for logarithmic growth phase of Jurkat cells.PT-PCR detected variety in CXCR4 mRNA,and cell cycle and apoptosis rate were analysed by flow cytometry (FCM).Results:Expression level of CXCR4 mRNA on transfected-Jurkat cells had a significant decine,compared with the blank control group.Cell proportion of G0/G1 cell cycle was increase,G2/M and S cell cycle was decreased,Apoptotic cells were significantly increased.Conclusion:Regrouping luciferase reporter vector (PgL4.17-CXCR4) was transfected into Jurkat cells,which decreased CXCR4 gene effectively,induced Jurkat cells apoptosis and blocking cell cycle,inhibited cell proliferation.%目的:探讨大蒜素转染后下调CXCR4表达对Jurkat细胞周期和凋亡的影响及大蒜素的作用机制.方法:克隆人CXCR4启动子序列,构建报告载体PgL4.17-CXCR4,转染Jurkat细胞,用不同浓度的大蒜素处理对数生长期的Jurkat细胞,PT-PCR检测CXCR4 mRNA的变化水平,流式细胞仪检测细胞周期和凋亡.结果:不同浓度剂量组与空白对照组相比,转染Jurkat细胞CXCR4 mRNA表达水平明显下降,G0/G1期细胞比例增加,G2/M期和S期细胞比例相应下降,凋亡细胞明显增加.结论:重组报告载体PgL4.17-CXCR4转染Jurkat细胞,能够有效下调CXCR4基因,诱导Jurkat细胞发生凋亡和细胞周期阻滞,从而抑制细胞增殖.