志苓胶囊通过激活caspase-3抑制K562细胞增殖和诱导细胞凋亡

The effects of Zhiling capsule on the proliferation inhibition and apoptosis induction in K562 cell line via caspase-3 pathway

目的研究志苓胶囊(ZLJN,抗癌复方Ⅱ号)对人慢性髓系白血病K562细胞株增殖及凋亡的影响。方法将志苓胶囊按其不同中西药成分比例配制成中药、西药和复方组,与K562细胞共培养后,采用MTT法、集落形成实验分别检测细胞存活率和集落形成率;Annexin V-FITC/PI标记法、DNA倍体分析及DNA片段化分析检测细胞凋亡;流式细胞仪检测caspase-3活性;Western blot法检测caspase-3酶原(pro-caspase-3)表达。结果不同药物组与K562细胞共培养后,细胞生长受抑制,集落形成率降低。Annexin V-FITC/PI法检测到早期凋亡细胞;DNA倍体分析可见亚二倍体峰(凋亡峰);琼脂糖电泳见典型的DNA梯状带。流式细胞检测caspase-3活性增强,Western blot检测pro-caspase-3表达减弱。结论志苓胶囊可有效抑制K562细胞增殖,诱导其凋亡,其作用机制可能与caspase-3活性增强有关。

Aim To investigate the effects of Zhiling capsule （ZLJN） on the proliferation inhibition and apoptosis induction in K562 cell line. Methods According to the different components of ZLJN, K562 cells were treated respectively with tradtional Chinese medicine, Western medicine and ZLJN compound groups. The cell viability and colony formation were observed by MTT assay and colony formation assay respectively. Apoptotic cells were detected by Annexin V-FITC/PI staining and DNA fragmentation assay. Caspase-3 activity was detected by flow cytometry, and pro-caspase-3 was detected by Western blot. Results Treated with drug,KS62 cell growth and cell colony formation were significantly inhibited. Apoptosis occurring in the early stage was identified by Annexin V-FITC/PI staining. Typical DNA ladder was seen from gel eleetrophoresis and apparent apoptotic peaks were observed by flow cytometer. The level of caspase-3 activity increased after the treatment, while the level of pro-caspase-3 decreased. Conclusion ZLJN can efficiently inhibit proliferation and induce apoptosis in K562 cells, which may be related with the up-regulation of caspase-3 activity.