1,25-二羟维生素D3对人胃黏膜上皮细胞恶性转化过程的影响  

Effect of 1,25-dihydroxy vitamin D3 on GES-1 during malignant transformation

作　　者：张明明[1,2] 贾彬[1] 陈栋[1] 闫慧明[1] 

机构地区：[1]山西医科大学第二临床学院普外科,太原030001 [2]内蒙古准格尔旗中心医院普外科

出　　处：《山西医科大学学报》2013年第6期435-438,共4页Journal of Shanxi Medical University

基　　金：山西省卫生厅科技基金资助项目(200920)

摘　　要：目的研究1,25-二羟维生素D3(骨化三醇)在化学致癌剂N-甲基-N-硝基-N-亚硝基胍(MNNG)诱导的恶性转化过程中对人胃黏膜上皮细胞系GES-1细胞生长、凋亡及Shh、Gli1基因表达的影响。方法 MNNG诱导GES-1细胞作为对照组,诱导后用不同浓度(10-6,10-7,10-8,10-9mmol/L)1,25-二羟维生素D3干预(实验组),采取四甲基固氮唑盐(MTT)检测实验组和对照组细胞增殖并确定实验组最佳药物浓度。分为GES-1细胞组、对照组和实验组,流式细胞术(FCM)检测各组细胞的凋亡率,RT-PCR检测各组细胞Shh、Gli1 mRNA基因的表达。结果 MTT法显示,对照组细胞出现大量死亡,实验组给予10-6,10-7,10-8,10-9mmol/L的1,25-二羟维生素D3干预后存活细胞较对照组增多(P<0.05)。取最佳药物作用浓度10-6mmol/L为实验组,FCM测各组细胞7 d的凋亡率分别为实验组(45.73±2.08)%、对照组(85.23±2.08)%、GES-1细胞组(4.9±0.91)%。与对照组相比,实验组细胞凋亡明显减少(P<0.05)。RT-PCR显示Shh mRNA在实验组(0.674 7±0.011 6)、对照组(0.661 6±0.009 5)均表达(P>0.05),Gli1 mRNA实验组(0.338 2±0.015 0)表达低于对照组(0.532 8±0.011 5)(P<0.05)。结论 1,25-二羟维生素D3有阻止GES-1细胞恶性转化的作用,从而可达到延缓或阻止细胞恶变的目的。其机制可能与通过阻断Sonic Hedgehog(SHH)信号通路有关。Abstract： Objective To explore the effect of 1,25-dihydroxy vitamin D3（calcitriol）on cell growth,apoptosis and Shh,Gli1 gene expression in in vitro cultured human gastric epithelial cell line GES-1 in the process of N-methyl-N-nitro-N-nitrosoguanidine （MNNG）-induced malignant transformation.Methods GES-1 cells were cultured with MNNG （control group）,and then were interfered with different concentrations of 1,25-dihydroxy （10^-6,10^-7,10^-8,10^-9 mmol/L） vitamin D3 （experimental groups）,respectively.MTT assay was used to detect the cell proliferation and screen the optimized concentration.The cells were divided into GES-1 group,control group and experimental group（optimized concentration）.Flow cytometry（FCM） was used to detect the cell apoptosis.RT-PCR was applied to determine the Shh,Gli1 mRNA expression.Results MTT results showed that there was a large number of cell death in control group.The survival rates in experimental groups were significantly higher than in control group.The optimized concentration was 10^-6 mmol/L in experimental groups.The apoptosis rate was （45.73 ± 2.08）% in experimental group,（85.23 ± 2.08）% in control group,and （4.9 ±0.91）% in GES-1 group.Compared with control group,the apoptosis reduced significantly in experimental group （P 〈 0.05）.RT-PCR showed that SHH mRNA was 0.674 7 ± 0.011 6 in experiment group and 0.661 6 ± 0.009 5 in control group （P 〉 0.05）.Gli1 mRNA expression in experiment group （0.338 2 ± 0.015 0）was lower than in control group（0.532 8 ± 0.011 5,P 〈0.05）.Conclusion 1,25-dihydroxy vitamin D3 can prevent the malignant transformation of GES-1 cells to delay or prevent the formation of malignant cells,which may be related to the block of the Sonic Hedgehog（SHH） signaling pathway.

关 键 词：1 25-二羟维生素D3 人胃黏膜上皮细胞 细胞凋亡 恶变 

分 类 号：R363[医药卫生—病理学]