雌二醇对子宫内膜癌细胞AKT通路中细胞因子的影响

通过17β雌二醇（E2）诱导人子宫内膜癌HEC-1A细胞产生不同细胞因子，探讨雌激素在子宫内膜癌发展中的作用。方法:以浓度为1×10-6 mol/L雌二醇（E2组）作用于HEC-1A细胞8h、12 h后，或1×10-6 mol/L雌激素受体抑制剂（ER组）、25×10-6 mol/L AKT抑制剂（AKT组）分别预处理HEC-1A细胞60min，各加入1×10-6 mol/L雌二醇作用8h和12h后；采用荧光定量PCR及ELISA技术，分别检测细胞内VEGF、bFGF、IL-8基因mRNA及细胞培养上清液中蛋白表达情况。Western Blot检测以1×10-6 mol/L雌二醇作用HEC-1A细胞15min后，细胞内AKT蛋白表达情况。结果:E2组的VEGF、bFGF、IL-8 mRNA及蛋白的表达均明显高于对照组（P<0.05）；ER组VEGF、bFGF、IL-8 mRNA和蛋白的表达，与E2组相比较，除了8h的VEGF蛋白和12 h的IL-8 mRNA表达无显著性差异及12 h的VEGF mRNA表达稍增加外，均明显低于E2组（P<0.05）；AKT组VEGF、bFGF、IL-8 mRNA和蛋白的表达，与E2组相比较，除了12 h的IL-8 mRNA表达无显著性差异外，均明显低于E2组（P<0.05）。雌二醇作用HEC-1A细胞15 min后，与对照组比较，细胞内p-AKT蛋白表达明显增强（P<0.05）。结论:雌二醇诱导子宫内膜癌产生细胞因子VEGF、bFGF、IL-8可能是通过激活AKT通路实现的。 

Effect of Estrogen on the Cytokines of AKT Signaling Pathway in Endometrial Cancer Cells

To investigate the relationship between estrogen and the development of endometrial cancer. Methods: Real-time polymerase chain reaction ( PCR ) was used to detect VEGF, bFGF, and IL-8 mRNA expression in HEC-1A cells and an enzyme-linked immunosorbent assay ( ELISA ) was used to detect the VEGF, bFGF, and IL-8 proteins in the cell culture fluid after stimulation with 1 × 10-6 mol/L estradiol ( E2 group ) for 8 h or 12 h, and with 60 min pretreatment with 1 × 10-6 mol/L ER inhibitor ( ER group ) or 25 × 10-6 mol/L AKT inhibitor ( AKT group ) following stimulation with 1 × 10-6 mol/L estradiol for 8 h or 12 h. Western blot was used to detect AKT protein expression in the HEC-1A cells after stimulation with 1 × 10-6 mol/L estradiol for 15 min. Results: Real-time PCR and ELISA showed the mRNA and protein expression of VEGF, bFGF, and IL-8 in the E2 group was significantly higher than that in the control group ( P < 0.05 ). The mRNA and protein expression of VEGF, bFGF, and IL-8 in the ER group was significantly lower than that in the E2 group ( P < 0.05 ) except for the VEGF protein expression at 8 h and the IL-8 mRNA expression at 12 h. VEGF mRNA expression was slightly high in the E2 group at 12 h. The mRNA and protein expression of VEGF, bFGF, and IL-8 in the AKT group was significantly lower than that in the E2 group ( P < 0.05 ), except for IL-8 mRNA expression at 12 h. Western blot analysis showed that p-AKT protein expression in the HEC-1A cells after 15 min stimulation with 1 × 10-6 mol/L estradiol was markedly higher than that in the control group ( P < 0.05 ). Conclusion: Estrogen induced the production of VEGF, bFGF, and IL-8 through activating the AKT signal transmission pathway.