雌激素受体在肺腺癌A549细胞系EMT过程中的作用

The Role of Estrogen Receptor in EMT of Lung Adenocarcinoma Cell Line A549

目的:探讨何种雌激素受体在肺腺癌A549细胞系上皮间质转化(EMT:Epithelial-mesenchymal transition)过程中发挥作用。方法:用雌激素受体拮抗剂分别抑制A549细胞系中的雌激素和受体,再用浓度为1×10^(-8)mol/L的雌激素刺激抑制前后的各组细胞系,用q-RT-PCR及Western blot实验检测各组细胞中EMT标志物的表达量,用Transwell实验检测各组细胞迁移量,计算各组间有无统计学差异。结果:雌激素组细胞中Vimentin的mRNA表达量为5.81±0.71,显著高于空白对照组,E-cadherin的mRNA表达量为0.43±0.07,显著低于空白对照组,细胞迁移数量为80.17±8.82,显著高于空白对照组,组间有统计学差异。用雌激素受体拮抗剂MPP处理前后Vimentin的mRNA表达量分别为7.63±0.92和6.29±0.73,均显著高于空白对照组,组间有统计学差异;E-cadherin的mRNA表达量分别为0.39±0.05和0.42±0.06,均显著低于空白对照组,组间有统计学差异;细胞迁移数量分别为85.21±11.56和78.69±9.63,均显著高于空白对照组,组间有统计学差异。雌激素加雌激素受体拮抗剂PHTPP组及雌激素加雌激素和受体拮抗剂组Vimentin的mRNA表达量分别为1.21±0.15和1.17±0.13,E-cadherin的mRNA表达量分别为0.86±0.12和0.93±0.11,细胞迁移数量分别为39.85±5.22和37.21±4.95,与对照组无统计学差异。各组细胞中EMT标志物蛋白表达量与mRNA表达量趋势相同。结论:雌激素β受体是雌激素促进A549细胞系发生EMT过程的重要靶点,在肺腺癌的肿瘤发生和发展过程中发挥了主要作用,可以作为进一步靶向治疗的研究方向。

Objective:To investigate the role of estrogen receptor in epithelial mesenchymal transition(EMT)of lung adenocarcinoma cell line A549.Methods:Estrogen receptor antagonists were used to inhibit estrogen receptor in A549 cell line,and then estrogen of 1×10^(-8) mol/L concentration was used to stimulate the cell lines before and after inhibition.The expression of EMT markers in each group was detected by q-RT-PCR and Western blot,and the cell migration rate of each group was detected by Transwell,and the statistical difference between each group was calculated.Results:The m RNA expression of vimentin in estrogen group was 5.81±0.71,which was significantly higher than that in blank control group.The m RNA expression of E-cadherin in estrogen group was 0.43±0.07,which was significantly lower than that in blank control group.The number of cell migration in estrogen group was 80.17±8.82,which was significantly higher than that incontrol group.The m RNA expression of vimentin were 7.63±0.92 and 6.29±0.73 before and after treatment with MPP,which were significantly higher than those in control group(P<0.05);the m RNA expression levels of E-cadherin were0.39±0.05 and 0.42±0.06,which were significantly lower than those in the control group;the number of cell migration were 85.21±11.56 and 78.69±9.63 before and after treatment with MPP,which were significantly higher than that of the control group,and there was statistical difference between the two groups.The m RNA expression of vimentin was 1.21±0.15 and 1.17±0.13,the m RNA expression of E-cadherin was 0.86±0.12 and 0.93±0.11,and the number of cell migration was 39.85±5.22 and 37.21±4.95 in estrogen plus PHTPP group and estrogen plus MPP and PHTPP group respectively which were no significant difference with control group.The trend of EMT marker protein expression and m RNA expression in each group was the same.Conclusion:Estrogen receptor β is an important target for estrogen to promote the EMT process of A549 cell line.It plays an important role in the carcinogenesis and development of lung adenocarcinoma,and can be used as the research direction of further targeted therapy.