维生素D_3对EC9706细胞增殖、裸鼠移植瘤生长及NDRG1蛋白表达的影响

Effect of Vitamin D_3 on EC9706 cells proliferation,xenograft tumor growth and expression of NDRG1 protein

目的:观察1,25-(OH)2维生素D3对食管癌EC9706细胞细胞增殖、细胞周期、食管癌细胞裸鼠移植瘤生长及对NDRG1蛋白表达的影响。方法:采用0.3μmol/L的1,25-(OH)2维生素D3作用于EC9706细胞后,MTT法检测肿瘤细胞的增殖情况,流式细胞仪检测细胞周期的变化。BALB/c小鼠荷瘤后,用5μg/kg1,25-(OH)2维生素D3溶液皮下注射于移植瘤周围,观察移植瘤生长情况,并采用免疫组织化学方法检测裸鼠移植瘤组织中NDRG1蛋白表达水平。实验中均以未经1,25-(OH)2维生素D3处理作为对照组。结果:①MTT实验结果显示,1,25-(OH)2维生素D3作用1～5d时各组吸光度均低于对照组,差异有统计学意义(P<0.05);随着1,25-(OH)2维生素D3作用时间的延长,细胞增殖抑制率逐渐升高(1d:7.4%,3d:21.1%,5d:23.8%)。②细胞周期检测结果显示,不同培养时间组G1期细胞分别与对照组比较,差异均有统计学意义(P<0.05)。③与对照组相比,1,25-(OH)2维生素D3可上调NDRG1蛋白表达水平。结论:1,25-(OH)2维生素D3可抑制食管癌细胞增殖,并通过上调NDRG1的表达而抑制食管癌裸鼠移植瘤的生长。

Aim：To study the proliferation,cell cycle distribution, expression of NDRG1 protein and xenografi tumor growth of human esophageal carcinoma EC9706 cells after being treated with 1,25-（OH） 2 Vitamin D3. M ethods：EC9706 cells were treated with 0.3 μmol/L 1,25-（ OH）2 Vitamin D3 ,then cell proliferation was detected by MTT method and cell cycle distribution was observed by flow cytometer. The xenograft tumor was therapied by 5 μg/kg 1,25- （OH）2 Vitamin D3 and the expression of NDRG1 protein in xenografts tumor was detected by immunohistochemistry method. Results： After EC9706 cells were exposed to 1,25-（OH） 2Vitamin D3 for 1 - 5 d, the mean absorbance values at 490 nm at different time points were lower than those in control group（ P 〈0.05 ）. The inhibition rate of proliferation at different time was ：7.4% at day 1, 21.1% at day 3, 23.8% at day 5 ,respectively. The inhibition rate increased with time. Flow cytometry （FCM） revealed that after EC9706 cells being exposed to 1,25-（OH） 2 Vitamin D3 for 1 -5 d, the cell cycle distribution changed significantly. There were significant differences in the percentages of the cells in G1 phase between experiment group and control group （P 〈 0.05 ）. 1,25-（ OH）2 Vitamin D3 could upregulate the expression of NDRG1 protein. Conclusion： 1,25- （OH） 2 Vitamin D3 may inhibit growth of xenograft tumor through regulating expression of NDRGI.