阿奇霉素诱导淋巴细胞白血病Jurkat细胞凋亡作用的研究

Azithromycin induced apoptosis of human T-cell lymphocyte leukemia cell Jurkat in vitro

目的:研究阿奇霉素诱导淋巴细胞白血病Jurkat细胞凋亡作用,及其相关的凋亡蛋白和细胞周期的变化,为临床治疗淋巴细胞白血病提供新的研究思路。方法:应用Annexin V-FITC/PI双染法流式细胞术检测细胞凋亡并用流式细胞术进行细胞周期分析,MTT法检测阿奇霉素对Jurkat细胞的增殖抑制作用,利用West-ern-blotting检测凋亡途径相关蛋白的表达。结果:用阿奇霉素处理Jurkat细胞48h后300mg/L剂量组早期凋亡率显著增高达到(88.0±3.3)%,而25mg/L剂量组为(11.5±2.7)%,与对照组(10.6%±1.8)%无明显差异。MTT法检测结果IC50值为88.4mg/L,200mg/L抑制率达到89.27%。细胞周期显示150mg/L时G1期细胞增加,G2期和S期细胞下降。Western-blotting结果表明Bcl-2蛋白高表达,处理前后无明显变化,而Bax蛋白表达随药物浓度提高而上升,活化的Caspase3在100mg/L表达最高。结论:阿奇霉素可以有效的诱导Jurkat细胞凋亡并呈明显的浓度依赖性,其凋亡途径可能是通过Bax蛋白表达上升诱导下游效应分子Caspase3活化实现。细胞增殖抑制试验表明200mg/L以上时细胞增殖明显受抑制,细胞周期分析提示细胞增殖阻滞在了G1期。

Objective：To observe the role of Azithromycin in apoptosis and cell cycle of human T-cell lymphocyte leukemia Jurkat cell in vitro.Method：Apoptosis was detected by Annexin V-FITC/PI kit through flow cytometry.Cell cycle was also analyzed by flow cytometry.Inhibitive effect of Azithromycin on cell growth was determined by MTT test.Expression of apoptosis protein was examined by western-blotting.Result：After 48 hours treatment with Azithromycin,the early apoptosis cells greatly increased in high concentration group but did not obviously change in low concentration group compared with control group.IC50 was 88.4 mg/L in MTT test and inhibition rate was 89.27% at 200 mg/L Azithromycin.Cell cycle analysis indicated the number of the cells increased in the G1 phase but decreased in the S phase and G2 phase.Western-bolting showed expression of anti-apoptosis protein Bcl-2 was unchanged and pro-apoptotic protein Bax was increased.Active-caspase3,which is downstream effector of apoptosis,was with increased expoession.Conclusion：Azithromycin could inhibit cell growth at high concentration and induced apoptosis of human T-cell lymphocyte leukemia cell in vitro.Cell cycle analysis revealed cell growth was blocked at G1 stage.Western-blotting suggested that apoptosis was related to increasing Bax protein expression and inducing Caspase3 activation.