氟维司群对亚砷酸钠致人支气管上皮细胞恶性转化的抑制作用研究

目的 研究雌激素受体拮抗剂氟维司群对亚砷酸钠致人支气管上皮细胞恶性转化的抑制作用，为探讨雌激素受体在砷致肺癌发生中的作用提供实验依据。方法 单纯以2.5 μM亚砷酸钠对人支气管上皮细胞株16-HBE持续染毒者设为对照组，自染毒30代起加入不同剂量（10-11、10-9、10-7M）氟维司群与砷联合处理者设为低、中、高剂量拮抗剂组，同时设置溶剂对照组（DMSO）。各组细胞继续培养至60代时，利用qPCR和Western Blot测定氟维司群干预下细胞ERα、ERβ mRNA和蛋白的相对表达水平，软琼脂克隆形成实验和划痕实验分别测定氟维司群对细胞克隆形成能力和迁移能力的影响。结果 培养至60代时，各剂量拮抗剂组细胞ERα mRNA和蛋白表达均低于对照组和溶剂对照组(P＜0.05)；中、高剂量拮抗剂组细胞ERβ mRNA表达均低于对照组和溶剂对照组(P＜0.05)，各剂量拮抗剂组细胞ERβ 蛋白表达水平均低于对照组和溶剂对照组(P＜0.05)。各剂量拮抗剂组的软琼脂克隆形成数和细胞24 h迁移距离均低于对照组和溶剂对照组(P＜0.05)。结论 氟维司群通过下调雌激素受体表达抑制了亚砷酸钠诱导的人支气管上皮细胞株16-HBE恶性转化，雌激素受体可能参与了砷致细胞恶性转化。

Inhibition effect of fulvestrant on the sodium arsenite-induced malignant transformation of human bronchial epithelial cells

Objective The aim of this study was to evaluate the inhibition effect of estrogen receptor antagonist fulvestrant on the sodium arsenite-induced malignant transformation of human bronchial epithelial cell, and provide experimental reference for determining the role of estrogen receptors in the development of lung cancer induced by arsenite. Methods We set control group(human bronchial epithelial cells 16-HBE were exposed to 2.5 μM NaAsO2 only ), low, middle, high dose fulvestrant group (cells were treated with 2.5 μM NaAsO2 and 10-11, 10-9, 10-7M fulvestrant after exposed to NaAsO2 for 30 passages) and solvent control group(DMSO). After culturing to 60 passage, qPCR and western blot were used to detect expression of ERα, ERβ mRNA and protein in 16-HBE under fulvestrant treatment. Soft agarose colony formation assay and scratch wound healing assay were conducted to determine the effects of fulvestrant on cell colony forming ability and migration ability, respectively. Results At 60 passage, expression of ERα mRNA and protein in 16-HBE of all-dose fulvestrant groups were lower than control and solvent control group(P<0.05). Expression of ERβ mRNA of middle and high-dose fulvestrant groups were lower than control and solvent control group(P<0.05). Expression of ERβ protein of all-dose fulvestrant groups were lower than control and solvent control group (P<0.05). The colony numbers formed in soft agarose and cell migration distance in 24 h of all-dose fulvestrant groups were lower than control and solvent control group(P<0.05). Conclusion Fulvestrant inhibited the sodium arsenite- induced malignant transformation of human bronchial epithelial cells(16-HBE) mediate by deregulating the expression of estrogen receptors. Estrogen receptors may involve in the arsenite-induced cell malignant transformation.