新型鬼臼毒素衍生物诱导人肺腺癌A549细胞凋亡及其机制研究  

Mechanisms Investigation of A Novel Podophyllotoxin Derivative In- duces Apoptosis in Lung Adenocarcinoma Cell Line A549

作　　者：王晓辉[1] 李佳蔚[1] 罗芸[2] 王剑锋[1] 惠玲[1] 

机构地区：[1]兰州军区兰州总医院医学实验中心甘肃省干细胞与基因药物重点实验室,兰州市730050 [2]兰州军区兰州总医院干部病房,兰州市730050

出　　处：《医学分子生物学杂志》2012年第3期183-188,共6页Journal of Medical Molecular Biology

基　　金：国家自然科学基金（No.30400125）,甘肃省自然科学基金（No.1107RJZA106）

摘　　要：目的探讨新型鬼臼毒素衍生物10Ⅲg诱导人肺腺癌细胞A549细胞凋亡及其调控机制。方法采用四甲基偶氮唑蓝（MTr）比色法、流式细胞术测定细胞周期细胞凋亡率，DNA琼脂糖凝胶电泳和微管蛋白组化染色，Western印迹法检测凋亡蛋白Bax、Caspase-3的表达。结果新型鬼臼毒素衍生物10m。对A549细胞增殖具有明显的剂量和时间依赖性抑制作用，细胞周期分析显示S期细胞数明显增多，出现G2／M期阻滞；10Ⅲg作用48h后DNA电泳可见明显的梯状条带；10Ⅲg能破坏A549细胞的细胞骨架，与依托泊苷相比有明显促进微管解聚现象；10Ⅲg浓度为10^-6mol/L时能显著促进Bax、Caspase-3蛋白的表达。结论新型鬼臼毒素衍生物10Ⅲg通过诱导A549细胞发生G2／M期阻滞而抑制其增殖，其机制可能与抑制细胞微管解聚及诱导细胞凋亡有关。Objective To investigate the effects of apoptosis on human lung adenocarcinoma A549 cells induced by a new derivative of podophyllotoxin 10Ⅲg and its mechanisms. Methods MTT assay, flow eytometry, DNA agarose gel electrophoresis and cell cycle analysis were used to detect apoptosis in A549 cells. Expression of Bax and Caspase-3 were measured with western blot. Results 1010Ⅲg was demonstrated to exert antiproliferative activity in a dose- and time-dependent manner. Cell cycle analysis indicated an increased S phase proportion, as well as an apparent G2/M phase arrest. After 48 h exposure with10Ⅲg, agarose gel electrophoresis showed an evident DNA fragmentation. 10Ⅲg could destroy the cytoskeleton of A549 cells, and significantly improve microtu- bule depolymerization compared with etoposide. The treat group of 10rmoL/L 10Ⅲg dramatically in- creased the expression of Bax and Caspase-3. Conclusion 10Ⅲgscould inhibit the growth of A549 cells via the induction of G2/M phase arrest, which is probably associated with the inhibition of tu- bulin polymerization and apoptosis induction.

关 键 词：鬼臼毒素 A549细胞 细胞凋亡 微管蛋白 Bax Caspase-3 

分 类 号：Q28[生物学—细胞生物学]