CD437诱导HepG2细胞凋亡及机制的初步研究

目的 探讨CD437对HepG2细胞凋亡诱导作用及其对线粒体膜电位变化的影响.方法 ①MTT比色法观察CD437对HepG2细胞增殖的影响,倒置显微镜下观察细胞生长情况.An-nexin V-FITC/PI标记后,流式细胞术检测细胞凋亡.②罗丹明123(Rhodamine123,Rh123)荧光探针染色后,荧光显微镜下观察活细胞线粒体改变,并经流式细胞仪分析线粒体膜电位变化.结果 ①1 μmol/L CD437干预HepG2细胞24h后,实验组细胞早期凋亡率(18.26±5.31)%高于对照组(P<0.05),实验组细胞继发死亡率(20.31±3.70)%也高于对照组(P<0.05),CD437可导致细胞早期凋亡.②流式细胞仪结果表明CD437诱导HepG2细胞24 h后,实验组荧光强度为4.40±0.09,较对照组荧光强度5.34±0.12变弱(n=12,P<0.05),实验组整个峰向左移动,HepG2细胞线粒体膜电位水平下降.荧光倒置显微镜下观察发现,对照组罗丹明123荧光强度很强,大量细胞发出强烈荧光,实验组HepG2细胞内罗丹明123荧光强度很弱,很少见到强荧光细胞.结论 CD437可能通过降低线粒体膜电位水平而抑制HepG2细胞增殖并且诱导其凋亡.

Effect and mechanical study of CD437 on apoptosis of human HepG2 cells

Objective To investigate the effect of CD437 on apoptosis of human hepatoblastoma cell line HepG2 and the possible role of CD437 on mitochondrial membrane potentials (MMP). Meth-ods The effect of CD437 on the proliferation of HepG2 cells was evaluated with MTT assay. Flow cytometry was performed after HepG2 cells were labeled with Annexin V-FITC/PI double fluores-cence. Mitochondrial membrane stained with Rhodamine 123 probes was detected under fluorescent microscope and the changes of MMP were analyzed with flow cytometry. Results After incubated with 1μmol/L of CD437 for 24 hours, both the earlier and later apoptotic rates of HepG2 cells in the exper-imental group increased significantly compared to those in the control group. The fluorescent intensity detected with flow cytometry in the experimental group was significantly lower than that in the control group (5. 34 ± 0. 12 vs 4. 40 ± 0. 09, P<0. 05). The left peak-shift and decreased MMP were noted in the experimental group. Under the fluorescent microscope, fluorescence of HepG2 stained with Rho-damine 123 probes disappeared after incubated with CD437. Conclusions CD437 can significantly in-hibit the growth of HepG2 and induce the apoptosis by decreasing MMP.