摘要
目的研究志苓胶囊(ZLJN)对人小细胞肺癌细胞系NCI-H446增殖影响及诱导凋亡作用。方法将志苓胶囊按其不同的中西药成分配制成不同浓度的中药组、西药组和复方组,与NCI-H446共培养后,采用MTT比色法、集落形成试验,分别检测细胞存活率和集落形成率。通过流式细胞仪进行DNA倍体分析、Bcl-2蛋白表达、线粒体跨膜电位及Caspase-3活性检测;DNA片段化分析法、Annexin V-FITC标记法、TdT酶介导的原位缺口末端标记法(TUNEL)检测凋亡细胞。结果不同浓度的药物组与NCI-H446共培养后,细胞生长明显受抑制,细胞集落形成率明显降低,呈浓度依赖性;各药物组处理后的细胞线粒体跨膜电位和Bcl-2蛋白表达水平下降,Caspase-3活性增强;琼脂糖凝胶电泳可见典型的DNA梯形带;各药物组均检测到明显的亚二倍体G1峰(凋亡峰);Annexin V-FITC法检测到早期凋亡细胞;TUNEL法检测到晚期凋亡细胞。复方组诱导细胞凋亡作用强于中药组和西药组。结论志苓胶囊可以有效抑制NCI-H446细胞增殖,诱导其凋亡,细胞线粒体跨膜电位水平降低,Caspase-3活性增强和Bcl-2表达下调可能参与了该过程。
Objective To investigate the effects of Zhiling Capsule （ZLC） on the proliferation inhibition and apoptosis induction in human small cell lung cancer cell line NCI-H446. Methods According to the different components of ZLC, NCI-H446 cells were treated with traditional Chinese medicine, Western medicine and ZLC compound groups. The rates of cell viability and colony formation were observed by MTT assay and colony formation assay respectively. Cell cycle assay, Bcl-2 protein expression, chondrial transmembrane potential and Caspase-3 activity were detected by flow cytometer. Apoptotic cells were detected by DNA fragmentation assay, Annexin-V FITC staining and TUNEL labeling methods. Results After NCI-H446 cells were treated with various concentrations of drug groups, cell growth was significantly inhibited in a dose dependent manner. Cell colony formation was obviously lowered in the same way. The levels of chondrial transmembrane potential and Bcl-2 protein expression were decreased, while the levels of Caspase-3 activity were increased after the treatment. Typical DNA ladder were seen from gel electrophoresis, and apparent apoptotic peaks were observed by flow cytometer. Apoptosis occured in the early and late stage was identified by Annexin-V FITC staining and TUNEL labeling methods respectively. The ZLC compound group has stronger apoptosis induction than the other groups. Conclusion ZLC could efficiently inhibit growth and induce apoptosis in NCI-H446 cells, which may be related with the down-regulation of chondrial transmembrane potential and Bcl-2 protein expression and the up-regulation of Caspase-3 activity.
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2007年第6期531-534,共4页
Chinese Journal of Integrated Traditional and Western Medicine
基金
福建省科技重点资助项目(No.2004Y023)
