紫花牡荆素诱导人肝癌HepG2细胞凋亡的研究

Study on aoptosis of human hepatocellular carcinoma HepG2 cells induced by Casticin

目的研究紫花牡荆素(Casticin)对肝癌HepG2细胞增殖抑制和凋亡诱导的作用,并探讨其作用机制。方法用终浓度为0、0.5、1.0及2.0μmol/L的Casticin作用于HepG2细胞,于12、24及48h后采用MTT法检测细胞增殖抑制率;Hoechst33342核染色,观察细胞形态学变化;24h后收集各组肝癌HepG2细胞,流式细胞术检测细胞周期及凋亡率;RT-PCR检测survivinmRNA表达。结果 MTT法检测显示,Casticin对肝癌HepG2细胞有增殖抑制作用,并存在浓度和时间依赖关系;Hoechst33342染色后,可见核染色质凝集,凋亡细胞呈致密浓染。与对照组相比,Casticin处理后凋亡细胞比例增加。Casticin作用24h后,细胞被阻滞于G2/M期,随药物质量浓度的增加,细胞凋亡率逐渐增加。RT-PCR结果显示,Casticin下调肝癌HepG2细胞sur-vivinmRNA表达。结论 Casticin在体外对肝癌HepG2细胞有明显的增殖抑制和凋亡诱导作用,初步推断Casticin诱发肝癌细胞凋亡的作用与其对survivin基因表达的抑制有关。

[Objective]To study the effects of Casticin on the proliferation and apoptosis induction of human hepatocellular carcinoma HepG2 cells, and explore the mechanism. [Methods]Human hepatocellular carcinoma HepG2 cells were treated with 0, 0.5, 1.0, 2.0 μmol/l Casticin and the cell proliferation inhibition rates were determined by MTT assay 12 h, 24 h and 48 h after treatment. Morphological changes of cell nuclei were observed by Hoechst 33342 staining. In addition, cell apoptosis rates and cell cycle were determined by flow cytometry, and RTPCR was employed to detect the expression of survivin mRNA. [Results]It was demonstrated by MTT assay that the growth of HepG2 cells was inhibited by Casticin in a time and dose-dependent manner. The chromatine concentration and apoptotic cells with high concentrations were observed by Hoechst 33342 staining. The percentage of cells in G2/M phase increased, after treatment with Casticin , apoptosis rate of HepG2 cells increased with the increase of concentration （P 0.05）. It was indicated by RT-PCR that the expression of survivin mRNA in HepG2 cells treated with Casticin was downregulated. [Conclusion]Casticin has inhibitory effects on HepG2 cell proliferation and can induce cell apoptosis in vitro, which may be related to the inhibition of the expression of survivin mRNA.