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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 different concentrations of glucose and insulin on the expression of vascular endothelial growth factor (VEGF) mRNA in 4T1 tumor cells. Methods 4T1 tumor cells were cultured in vitro. 4T1 tumor cells were divided into 15 groups according to the concentration of glucose and insulin in RPMI 1640 medium. The cells were harvested at 37 ℃ and 5% CO2 for 48 h. The total RNA was extracted and RT-PCR was performed to amplify the target VEGF and β-actin simultaneously. Sepharose Electrophoresis, amplification products were scanned on a gel imaging system to calculate the relative expression of VEGF mRNA. The whole experiment was repeated 3 times. Results The expression of VEGF mRNA in B, C, D and E groups with different concentrations of glucose in culture medium did not change significantly (P> 0.05). The expression of VEGF mRNA in group F, group G and group J was not significantly different from that in group A (P> 0.05). In group H containing high concentration of insulin, the expression of VEGF mRNA in group I was significantly higher than that in group A The difference was statistically significant (P <0.05). The expression of VEGF mRNA in L group and M group was significantly lower than that in A group (P <0.05), but there was no significant difference between K group, N group and O group and A group (P> 0.05). Conclusion High concentration of insulin can up-regulate the expression of VEGF mRNA in 4T1 tumor cells. The change of glucose concentration has no effect on the expression of VEGF mRNA in 4T1 tumor cells.