大蒜素对重组质粒转染后Jurkat细胞周期和凋亡的影响

Effect of allicin on recombinant plasmid transfected Jurkat cell cycle and apoptosis

目的:探讨大蒜素转染后下调CXCR_4 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CXCR_4 启动子序列,构建报告载体Pg L4.17-CXCR_4 ,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CXCR_4 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CXCR_4 mRNA表达水平明显下降,G_0/G_1期细胞比例增加,G_2/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Pg L4.17-CXCR_4 转染Jurkat细胞,能够有效下调CXCR_4 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。

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 al-licin for logarithmic growth phase of Jurkat cells. PT - PCR detected variety in CXCR4 mRNA,and cell cycle and ap-optosis rate were analysed by flow cytometry （FCM）. Results： Expression level of CXCR4 mRNA on transfected - Jur- kat cells had a significant decine, compared with the blank control group. Cell proportion of G0/G ） cell cycle was in-crease, G2/M and S cell cycle was decreased, Apoptotic cells were significantly increased. Conclusion ： Regrouping lu-ciferase reporter vector （PgL4. 17 - CXCR4） was transfected into Jurkat cells,which decreased CXCR4 gene effective- ly,induced Jurkat cells apoptosis and blocking cell cycle,inhibited cell proliferation.