蟾毒灵对人非小细胞肺癌A549细胞增殖和凋亡的影响
[Effect of Bufalin on proliferation and apoptosis of human non-small cell lung cancer A549 cell].
作者信息
Zhu Zhitu, Yu Yunlong, Wang Kai, Li Enze, Liu Yangyang, Liu Yunpeng
机构信息
Department of Oncology, the First Affiliated Hospital of Liaoning Medical University, Jinzhou 121001, China.
出版信息
Zhongguo Fei Ai Za Zhi. 2010 Sep;13(9):841-5. doi: 10.3779/j.issn.1009-3419.2010.09.01.
BACKGROUND AND OBJECTIVE
Bufalin, a component of the Chinese medicine chan'su, induces apoptosis in various human tumor cell lines. The aim of this study is to investigate the mechanism of anti-proliferation and the induction of apoptosis of Bufalin on non-small cell lung cancer cell.
METHODS
The effect of Bufalin on A549 cell proliferation was measured using MTT assay; the morphological change was observed with Wright-Giemsa stain; cell cycle phase distribution and hypodiploid DNA were determined by flow cytometry with PI; expressions of Livin, Caspase-3 were detected by Western blot.
RESULTS
MTT assay showed that Bufalin inhibited A549 cell proliferation in time and dose-dependent manner. The inhibition concentration of 50% cell growth (IC50) at 48 h, 72 h and 96 h were (56.14+/-6.72) nmol/L, (15.57+/-4.28) nmol/L and (7.39+/-4.16) nmol/L, respectively; apoptosis of A549 cells could be effectively induced by Bufalin; apoptotic bodies and sub-G1 peak (representing apoptosis) emerged when cells were treated with different concentrations, compared with the untreated control group (P < 0.01). The expression of Livin protein was downregulated (P < 0.01) and Caspase-3 protein was activated during Bufalin-induced apoptosis in A549 cells.
CONCLUSIONS
Bufalin suppresses the growth of A549 cells and induces apoptosis; inhibition of Livin and activation of Caspase-3 may be involved in its mechanism.
背景与目的
蟾酥是中药蟾酥的一种成分，可诱导多种人类肿瘤细胞系发生凋亡。本研究旨在探讨蟾酥对非小细胞肺癌细胞的抗增殖及凋亡诱导机制。
方法
采用MTT法检测蟾酥对A549细胞增殖的影响；用瑞氏-吉姆萨染色观察形态变化；用PI通过流式细胞术测定细胞周期阶段分布和亚二倍体DNA；用蛋白质免疫印迹法检测Livin、Caspase-3的表达。
结果
MTT法显示蟾酥以时间和剂量依赖性方式抑制A549细胞增殖。48小时、72小时和96小时的50%细胞生长抑制浓度（IC50）分别为（56.14±6.72）nmol/L、（15.57±4.28）nmol/L和（7.39±4.16）nmol/L；蟾酥可有效诱导A549细胞凋亡；与未处理的对照组相比，用不同浓度处理细胞时出现凋亡小体和亚G1峰（代表凋亡）（P<0.01）。在蟾酥诱导A549细胞凋亡过程中，Livin蛋白表达下调（P<0.01），Caspase-3蛋白被激活。
结论
蟾酥抑制A549细胞生长并诱导凋亡；抑制Livin和激活Caspase-3可能参与其机制。
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