二甲双胍对体外子宫内膜癌细胞COX-2、VEGF和PTEN表达的影响

Effects of metformin on expression of cyclooxygenase-2,vascular endothelial growth factor,and phosphatase and tensin homolog deleted on chromosome ten in endometrial carcinoma cells in vitro

目的探讨二甲双胍在体外对人子宫内膜癌细胞环氧合酶-2(COX-2)、血管内皮生长因子(VEGF)和张力蛋白同源第10号染色体缺失的磷酸酶(PTEN)蛋白表达的影响。方法按照二甲双胍干预药物浓度的不同,将细胞实验分为实验组(0.01 mmol/L组,0.1 mmol/L组,1 mmol/L组,10 mmol/L组)和对照组(0 mmol/L组),分别干预4种人子宫内膜癌Ishikawa、RL-952、HEC-1A和KLE细胞相同时间后,采用ELISA法检测内膜癌细胞中COX-2和VEGF表达的变化,Western blotting法检测内膜癌细胞中PTEN蛋白的表达情况。结果Ishikawa和RL-952细胞中4个实验组相对于对照组COX-2和VEGF的表达均随着二甲双胍浓度的增加呈明显下降趋势,差异均有统计学意义(P<0.05);HEC-1A和KLE细胞中4个实验组相对于对照组COX-2的表达均随着二甲双胍浓度的增加呈下降趋势,但是仅有HEC-1A细胞中10 mmol/L组差异有统计学意义(P<0.05),其余实验组差异均无统计学意义(P>0.05);HEC-1A和KLE细胞中4个实验组相对于对照组VEGF的表达均随着二甲双胍浓度的增加呈明显下降趋势,差异有统计学意义(P<0.05);Western blotting法检测显示二甲双胍能够促进子宫内膜癌Ishikawa细胞中PTEN蛋白的表达,差异有统计学意义(P<0.05),而对RL-952、HEC-1A和KLE细胞中PTEN蛋白的表达无影响(P>0.05)。结论二甲双胍可能通过抑制子宫内膜癌细胞中COX-2与VEGF的表达来实现其部分的抗肿瘤作用。

Objective To investigate the effects of metformin on the expression of cyclooxygenase-2(COX-2),vascular endothelial growth factor(VEGF)and phosphatase and tensin homolog deleted on chromosome ten(PTEN)in human endometrial carcinoma cells in vitro.Methods According to the different concentrations of metformin intervention drugs,the cell experiments were divided into experimental group(0.01 mmol/L group,0.1 mmol/L group,1 mmol/L group,10 mmol/L group)and control group(0 mmol/L group).After the intervention of Ishikawa,RL-952,HEC-1A,and KLE cells of four kinds of human endometrial carcinoma for the same time,and the expression of COX-2 and VEGF in endometrial cancer cells was detected by ELISA.Expression of PTEN protein in endometrial cancer cells was detected by Western blotting.Results Compared with the control group,the expression levels of COX-2 and VEGF in the four experimental groups in the Ishikawa and RL-952 cell groups decreased significantly with the increase of metformin concentration,and the differences were statistically significant(P<0.05).Compared with the control group,the expression of COX-2 in the 4 experimental groups in the HEC-1A and the KLE cell group decreased with the increase of metformin concentration,but only the HEC-1A cell group had a statistically significant difference in the 10 mmol/L group(P<0.05),and there was no significant difference in other groups(P>0.05).In the HEC-1A and KLE cell,the expression of VEGF in the four experimental groups(compared with the control group)decreased significantly with the increase of metformin concentration,and the differences were statistically significant(P<0.05).Western blotting showed that metformin could promote the expression of PTEN protein in endometrial cancer Ishikawa cells,and the difference was statistically significant(P<0.05),but had no effect on the expression of PTEN in RL-952,HEC-1A and KLE cells(P>0.05).Conclusion Metformin partially achieves anti-tumor effects by inhibiting the expression of COX-2 and VEGF in endometrial cancer cells.