新型雌激素受体GPR30在非小细胞肺癌增殖中的作用研究

摘要

目的 研究G蛋白耦联受体30(GPR30)在非小细胞肺癌(NSCLC)中的表达及其与临床主要病理特征的关系,并分析GPR30和Ki-67表达的相关性,探讨雌激素通过激活GPR30受体信号途径调节NSCLC增殖的分子机制.方法 采用免疫组织化学方法检测80例术后非小细胞肺癌组织样本中GPR30和Ki-67的表达.加入17-β-雌二醇或G-1后,计数H1299细胞,流式细胞术检测细胞周期分布,最后通过Western blot方法检测G-1作用后ERK1/2的激活状态以及cyclin D1和P16蛋白的表达.结果 GPR30更多表达在腺癌、低分化、Ⅲ期NSCLC肿瘤组织,差异有统计学意义(P＜0.05);GPR30表达和Ki-67呈中度相关性(r=0.502,P=0.000).E2(或G-1)促进H1299细胞增殖,并且更多的细胞进入S期;加入G-1后,磷酸化ERK1/2以及cyclin D1表达增加,而p16蛋白表达减少,以上效应能被G-15或U0126预处理2h阻断.结论 雌激素通过激活GPR30-EGFR-MAPKs信号转导途径促进H1299增殖.阻断GPR30信号途径可能成为NSCLC治疗的新靶点.%Objective To evaluate the expression of GPR30 and Ki-67 in Non-small cell lung cancer(NSCLC) and the relationship between them.The clinicopathological features of GPR30 in NSCLC were also analyzed.The molecular mechanism that estrogen mediated the proliferation of H1299 by activating GPR30 was further studied.Methods The expression of GPR30 and Ki-67 in 80 cases of specimens of NSCLC after surgery was examined using immunohistochemistry method.After 17-β-estradiol(E2) or G-1 added,H1299 cells were counted and the cell cycle distribution was analyzed by flow cytometry.Finally,the activated ERK1/2 and the expression of cyclin D1 and p16 after G-1 treatment in H1299 cells were examined through western blotting.Results Expressions of GPR30 was more in stage Ⅲ or low differentiation tissues or adenocarcinoma (P＜0.05).A positive correlation between GPR30 and Ki-67 was further disclosed (r=0.502,P=0.000).The proliferation of H1299 cells was promoted and more cells entered S-p hase after E2 or G-1 treatment for 3 days,which could be inhibited after G-15 or U0126 pre-treatment for 2 hours.We further discovered that the activated ERK1/2 and cyclin D1 expression increased after G-1 treatment,which was blocked after G-15 or U0126 pre-treatment for 2 hours.The change of p16 was on the opposite.Conclusion A positive correlation existed between GPR30 and Ki-67.GPR30-EGFR-MAPKs signaling transduction pathway was involved in the estrogen-induced proliferation of NSCLC cells.Blocking GPR30 signaling pathway may be a promising new strategy for NSCLC treatments.