葡萄糖和胰岛素对colon26肿瘤细胞VEGF mRNA表达的影响

Effects of glucose and insulin on expression of VEGF mRNA in Colon26 cells

目的本实验模拟糖尿病病程不同阶段患者体内葡萄糖和胰岛素浓度的变化,研究不同浓度葡萄糖、胰岛素对colon26肿瘤细胞VEGF mRNA表达的影响。方法体外常规培养colon26肿瘤细胞,分别给予不同浓度葡萄糖、胰岛素进行干预,根据所用培养液外加葡萄糖和胰岛素浓度的不同,将实验所用细胞分为4组(12亚组),培养48 h,收集细胞后抽提总RNA,RT-PCR同时扩增目的片段VEGF和内参照β-actin,进行琼脂糖凝胶电泳,扩增产物在凝胶成像系统上扫描分析,计算VEGF mRNA的相对表达量。结果胰岛素浓度为125mU/L的亚组和其加入不同浓度的葡萄糖的亚组VEGF mRNA的表达明显高于对照组(P<0.01);其他亚组与对照组相比差异无统计学意义(P>0.05)。结论高浓度胰岛素能够上调colon26肿瘤细胞VEGF mRNA的表达,不同浓度的葡萄糖对colon26肿瘤细胞VEGF mRNA的表达无明显影响。

Objective To study the effects of glucose and insulin on the expression of VEGF mRNA in Colon26 cells by simulating the changes of glucose and insulin levels in different courses of diabetes mellitus. Methods Colon26 cells were cultured in vitro. According to the concentration of glucose and insulin in culture medium, the cells were divided into 4 groups（ 12 subgroups）. The cells were cultured for 48h, and total RNA was isolated after collecting cells and then the expression of VEGF mRNA was detected by reverse transcriptase-polymerase chain reaction （RT-PCR）. Results The expression of VEGF mRNA was significantly higher in 125 mU/L insulin than that in control group （P 〈 0.01 ）. There was no statistical difference in the expression of VEGF mRNA in Colon26 cells between the control groups and glucose/insulin subgroups except for 125 mU/L insulin （P 〉 0.05）. Conclusions High concentration of insulin may increase the expression of VEGF mRNA in Colon26 cells and the concentration of glucose has no effect on the expression of VEGF mRNA.