雌二醇调控VEGFα表达诱导肺腺癌A549细胞血管形成

Regulation of VEGFα expression by 17β-estradiol induced angiogenesis in lung adenocarcinoma

目的研究雌二醇(17β-estradiol,E2)在肺腺癌A549细胞微环境中,通过血管内皮生长因子α(vascular endothelial growth factorα,VEGFα)诱导肿瘤血管形成的发生机制。方法将A549细胞分为3组:PBS组、10 nmol·L^-1 E2组、10 nmol·L^-1 E2+5μmol·L^-1雌激素受体拮抗剂(ICI)组。Western blot和ELISA法检测肺腺癌A549细胞和培养基中VEGFα的表达;MTT法检测人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)的增殖能力;划痕实验检测HUVEC细胞迁移能力;小管生成实验检测HUVEC细胞的体外血管生成能力。结果与对照组(PBS)相比,E2刺激后的A549细胞中VEGFα的表达量增加,而加入ICI后,VEGFα表达量明显下降;同时,E2诱导A549细胞及上清液中VEGFα的表达,促进了HUVEC细胞的增殖、迁移和小管形成能力,ICI则相反。结论E2通过作用于A549细胞中雌激素受体,刺激肺腺癌微环境中VEGFα的表达,促进肺腺癌血管内皮细胞的增殖、迁移和小管形成能力,诱导肺腺癌血管形成。

Aim To investigate the mechanism of 17β-estradiol (E2) induced tumor angiogenesis by VEGFα in the microenvironment of lung adenocarcinoma in A549 cells. Methods A549 cells were divided into three groups: PBS group, 10 nmol·L ^-1 E2 group and 10 nmol·L ^-1 E2 +5 μmol·L^-1 estrogen receptor antagonist (ICI) group. Western blot was used to detect the expression of VEGFα in A549 cells, and ELISA was used to detect the expression of VEGFα in culture medium. The proliferation of human umbilical vein endothelial cells (HUVECs) was detected by MTT, the migration ability of HUVECs by scratch assay and the tubulogenesis in vitro by tube formation assay. Results Compared with control group, the expression of VEGFα in A549 cells in E2 group was higher, but the expression of VEGFα in A549 cells was significantly inhibited with the addition of ICI. Meanwhile, E2 induced the expression of VEGFα in A549 cells, promoting proliferation, migration and tubulogenesis of HUVEC, while ICI had the opposite effects. Conclusions E2 stimulates the expression of VEGFα by binding to estrogen receptor in A549 cells, promoting proliferation, migration and tube formation of HUVECs, contributing to lung adenocarcinoma angiogenesis.