骨化三醇体外诱导人肝癌细胞IGFBP-3的表达及细胞凋亡的实验研究

Effects of calcitriol on expression of insulin-like growth factor binding protein 3 and apoptosis of human liver carcinoma cell line

目的:探讨骨化三醇[2β-(3一hydro-)-calcitriol]对人肝癌细胞HepG2增殖和凋亡的影响及其作用机制。方法:应用MTT比色法观察骨化三醇对HepG2的生长抑制作用,以荧光显微镜和流式细胞仪观察HepGz的凋亡,RT-PCR检测骨化三醇处理前后HepG2细胞中胰岛素样生长因子受体结合蛋白-3(IGFBP- 3)mRNA表达水平的变化。结果:骨化三醇显著抑制HepG2细胞的体外增殖,并诱导细胞凋亡。不同浓度的骨化三醇(10-8,10-7,10-6,10-5mol·L-1)作用HepG2 48 h细胞凋亡率分别达(27.3±s 2．3)％,(33±6)％,(35±5)％,(48±7)％,明显高于对照组(8±4)％(P<0．05)。RT-PCR结果显示骨化三醇处理HepG2 24 h后IGFBP-3 mRNA表达水平明显升高。结论:骨化三醇体外显著抑制HepG2增殖并促进其凋亡,其作用机制可能与通过激活细胞内IGFBP-3 mRNA的表达有关。

AIM： To explore the effects of 2β-（3-hydroxyproxy） -calcitriol （ED-71） on proliferation, apoptosis, and induction of insulin-like growth factor binding protein 3 （IGFBP-3） in human liver carcinoma cell line HepG2. METHODS： HepG2 were treated with different concentrations of ED-71. MTT assay was employed to measure the proliferation of carcinoma cells. Epifluorescence microscopy was used to observe morphological changes in carcinoma cells and apoptosis was determined by flow cytometry. Expression of IGFBP-3 was determined by RT-PCR. The statistical analysis was performed using one-way ANOVA and student＇ s t test. RESULTS： ED-71 treatment led to a time- and dose-dependent inhibition in carcinoma cells HepG2 proliferation. ED- 71 caused an increase in cell apoptosis and the gene expression level of IGFBP-3. CONCLUSION： ED-71 could inhibit the proliferation in human liver carcinoma cell HepG2, which may be due to the expression of IGFBP-3.