丹酚酸C诱导肝癌HepG2细胞有丝分裂阻滞及凋亡的研究

Induction of mitotic arrest and apoptosis by salvianolic acid C in human hepatoma HepG2 cells

目的研究丹酚酸C(salvianolic acid C,SalC)的体外抗肿瘤细胞增殖作用及其分子机制。方法 MTT法检测Sal C对人肿瘤细胞的增殖抑制作用,流式细胞术检测细胞周期分布及细胞凋亡,Giemsa染色进行细胞有丝分裂的形态学观察,微管蛋白聚合实验检测Sal C对微管蛋白聚合活性的影响,比色法检测细胞中Caspase-3和Caspase-6的活性。结果丹酚酸C能抑制多种人肿瘤细胞增殖,其中对HepG2细胞的抑制作用较明显。流式细胞术分析发现Sal C使HepG2细胞阻滞于G2/M期并随作用时间延长出现明显的凋亡峰,细胞中Caspase-3和Caspase-6的活性升高。Gi-emsa染色提示HepG2细胞发生有丝分裂阻滞。Sal C能明显抑制微管蛋白的聚合。结论 Sal C具有抗肿瘤细胞增殖活性,通过抑制微管蛋白聚合诱导细胞有丝分裂阻滞从而诱导凋亡。

Aim To investigate the inhibitory effect of salvianolic acid C against human tumor cells and the related mechanisms.Methods In vitro growth inhibitory effect on a variety of tumor cell lines was determined by MTT assay.Cell cycle distribution and apoptosis were detected by flow cytometry.Giemsa staining was performed to demonstrate the morphological features of cells in mitotic phase.Polymerization of tubulin was detected by tubulin assembly assay.Activation of Caspase-3 and Caspase-6 was measured by colorimetric assay.Results MTT assay showed that salvianolic acid C inhibited the proliferation of several cancer cell lines,in which human hepatocellular cancer HepG2 cells exhibited the highest susceptibility to salvianolic acid C.HepG2 cells were markedly arrested in G2 /M phase and progressed into apoptosis after the treatment of salvianolic acid C.Increased activities of Caspase-3 and Caspase-6 in HepG2 cells were observed.Giemsa staining indicated mitotic blockade by salvianolic acid C.The drug also inhibited tubulin polymerization.Conclusions Salvianolic acid C displays antiproliferative effect on cancer cells.It blocks cell cycle through inhibiting tubulin polymerization and then induces apoptosis.