葡萄糖和胰岛素对4T1肿瘤细胞VEGF mRNA表达的影响

Effect of glucose and insulin on the expression of VEGF mRNA in 4T1 Cells

目的研究不同浓度的葡萄糖和胰岛素对4T1肿瘤细胞血管内皮生长因子(VEGF)mRNA表达的影响。方法体外常规培养4T1肿瘤细胞,根据细胞培养液RPMI 1640培养基中所含葡萄糖和胰岛素浓度的不同将4T1肿瘤细胞分为15组。每组设双复孔,置于37℃、5%CO2的细胞培养箱培养,48h后收集细胞,提取总RNA,RT-PCR同时扩增目的片段VEGF和内参照β-actin,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相对表达量。整个实验过程重复3次。结果培养液中含有不同浓度葡萄糖的B、C、D、E组VEGF mRNA的表达与A组相比变化不明显(P>0.05)。F组、G组和J组VEGF mRNA的表达与A组相比差异均无统计学意义(P>0.05);在含有高浓度胰岛素的H组、I组VEGF mRNA的表达水平与A组相比差异有统计学意义(P<0.05)。L组和M组的VEGF mRNA的表达水平与A组相比差异有统计学意义(P<0.05);而K组、N组和O组与A组相比差异均无统计学意义(P>0.05)。结论高浓度的胰岛素能够使4T1肿瘤细胞VEGF mRNA的表达上调,葡萄糖浓度的变化对4T1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 4T1 cells. Methods 4T1 tumor cells were cultured in vitro. According to the concentrations of glucose and insulin in culture medium, cells were divided into 15 groups. There were 2 repeated holes in each group. After differ- ent culture media was added, cells were cultured for 48h in saturated humidity incubator with 5% CO2 at 37℃ Expressions of VEGF were detected by RT - PCR and were repeated three times. Results Compared with group A, there was no statistical difference about the expression of VEGF mRNA in 4T1 ceils in B, C, D and E groups （P 〉 0. 05 ） . In group H and group I, the level of VEGF mRNA was higher significantly than A （P 〈 0. 05 ） . However, expression of VEGF in F, G and J groups had no difference compared with that in A group respectively （ P 〉 0. 05 ） . In group L and group M, the level of VEGF mRNA was higher significantly more than that of group A （ P 〈 0. 05 ） No differences were found in K, N and O groups compared with group A （P 〉 0.05） . Conclusions High concentration of insulin may up - regulate the expression of VEGF mRNA in 4T1 tumor cells. Different concentrations of glucose have no effect on the expression of VEGF mRNA in 4T1 tumor ceils.