三种去甲基斑蝥素衍生物体外对人肝癌HepG-2细胞增殖的抑制作用

目的:探讨去甲基斑蝥素酸钠、去甲基斑蝥素酸钾、去甲基斑蝥素酸钙体外对人肝癌HepG-2细胞增殖的影响。方法:采用磺酰罗丹明(SRB法)、细胞集落形成实验、流式细胞技术检测三种去甲基斑蝥素衍生物对HepG-2细胞增殖和凋亡的影响。qRT-PCR技术考察三种去甲基斑蝥素衍生物对HepG-2细胞Bcl-2、Bax mRNA表达的影响。结果:三种去甲基斑蝥素衍生物能明显抑制HepG-2细胞的增殖,其抑制率呈剂量时效依赖性;与空白对照组相比,三种去甲基斑蝥素衍生物能显著性减少HepG-2细胞集落形成数量,并且可诱导HepG-2细胞的凋亡,凋亡率分别为8±0.56%、6.93±0.91%、6.16±0.37%;qRT-PCR显示,三种去甲基斑蝥素衍生物能抑制HepG-2细胞Bcl-2mRNA的表达,增强Bax mRNA的表达。结论:三种去甲基斑蝥素衍生物可通过下调Bcl-2 mRNA表达,上调Bax mRNA表达,诱导HepG-2细胞的凋亡,从而抑制HepG-2细胞的增殖;三种衍生物中,去甲基斑蝥素酸钠对HepG-2细胞的增殖的抑制作用最强。

Inhibitory Effect of Three Norcantharidin Derivativeson HepG-2 Cells in Vitro

ABSTRACT Objective: To explore the inhibitory effect of three norcantharidin derivatives which were named sodium norcantharidate, potassium norcantharidate and calcium norcantharidate on human hepatocellular carcinoma HepG-2 cells. Methods: The effect of three norcantharidate derivatives on HepG-2 cells proliferation was assessed with SRB assay. The cell colony forming experiment test was adopted to observe the impact of derivatives on colony formation ability of HepG-2 cells. The flow cytometry was used to examine the effect on HepG-2 cellsapoptosis, and mRNA levels of Bcl-2 and Bax were measured with RT-qPCR. Results: Three norcantharidate derivatives inhibited the proliferation of HepG-2 cells with a time and dose dependent relation. Compared with the blank control group, derivatives were able to obviously inhibit cell colony formation and induce apoptosis of HepG-2 cells. The apoptosis rate were 8±0.56%, 6.93±0.91%, 6.16±0.37%, respectively. Base on the results of RT-qPCR, three norcantharidate derivatives had the ability to reduce the mRNA levels of Bcl-2 and improve the mRNA levels of Bax. Conclusion: The three kinds of norcantharidate derivatives could inhibit the proliferation and induce apoptosis of HepG-2 cells which involve in down-regulating the mRNA levels of Bcl-2 and up-regulating the mRNA levels of Bax. Among three derivatives, sodium cantharidinate shows the strongest inhibitory activity on HepG-2 cells.