Chen Ying-Yu, Pan Yun-Ling, Liu Ting-Bo
Fujian Institute of Hematology, Union Hospital of Fujian Medical University, Fuzhou.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2007 Jun;27(6):531-4.
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.
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.
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.
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.
探讨志苓胶囊(ZLC)对人小细胞肺癌细胞系NCI-H446增殖抑制及凋亡诱导的影响。
根据ZLC的不同成分,将NCI-H446细胞分为中药组、西药组和ZLC复方组进行处理。分别采用MTT法和集落形成试验观察细胞活力和集落形成率。通过流式细胞仪检测细胞周期、Bcl-2蛋白表达、线粒体跨膜电位和Caspase-3活性。采用DNA片段化分析、Annexin-V FITC染色和TUNEL标记法检测凋亡细胞。
用不同浓度药物组处理NCI-H446细胞后,细胞生长呈剂量依赖性显著抑制。细胞集落形成也明显降低。处理后线粒体跨膜电位和Bcl-2蛋白表达水平降低,而Caspase-3活性水平升高。凝胶电泳可见典型的DNA梯带,流式细胞仪观察到明显的凋亡峰。Annexin-V FITC染色和TUNEL标记法分别鉴定了早期和晚期发生的凋亡。ZLC复方组的凋亡诱导作用强于其他组。
ZLC可有效抑制NCI-H446细胞生长并诱导其凋亡,这可能与线粒体跨膜电位和Bcl-2蛋白表达下调以及Caspase-3活性上调有关。