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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 This experiment simulated the changes of glucose and insulin in patients with different stages of diabetes mellitus, and studied the effects of different concentrations of glucose and insulin on the expression of VEGF mRNA in colon26 tumor cells. Methods The colon26 tumor cells were cultured in vitro. The cells were divided into 4 groups (12 subgroups) according to different concentrations of glucose and insulin. The cells were harvested for 48 h. Cells were harvested After extraction of total RNA, RT-PCR was used to amplify VEGF and internal reference β-actin simultaneously, and then subjected to agarose gel electrophoresis. The amplified product was scanned and analyzed on a gel imaging system to calculate the relative expression level of VEGF mRNA. Results The expression of VEGF mRNA in subgroups with insulin concentration of 125 mU / L and those with different concentrations of glucose was significantly higher than that in control group (P <0.01). There was no significant difference between the other subgroups and the control group (P > 0.05). Conclusion High concentration of insulin can up-regulate the expression of VEGF mRNA in colon26 tumor cells. Different concentrations of glucose have no significant effect on the expression of VEGF mRNA in colon26 tumor cells.