肿瘤相关成纤维细胞中G蛋白偶联雌激素受体胞浆转位介导的旁分泌对乳腺癌细胞生长的影响  

Influence of paracrine mediated by cytoplasmic translocation of G protein-coupled estrogen receptor in cancer-associated fibroblasts on growth of breast cancer cells

作　　者：余腾骅 涂刚[2] 彭美茜 孙可馨 柳满然[3] YU Tenghua;TU Gang;PENG Meiqian;SUN Kexin;LIU Manran(Department of Breast Cancer Surgery, Jiangxi Cancer Hospital, Nanchang 330029, China;Department of Endocrine and Breast Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China;Key Laboratory of Laboratory Medical Diagnostics,Chinese Ministry of Education, Chongqing Medical University, Chongqing 400016, China)

机构地区：[1]江西省肿瘤医院乳腺肿瘤外科,江西南昌330029 [2]重庆医科大学附属第一医院内分泌乳腺外科,重庆400016 [3]重庆医科大学临床检验诊断学教育部重点实验室,重庆400016

出　　处：《中国普通外科杂志》2019年第5期573-580,共8页China Journal of General Surgery

基　　金：国家自然科学基金青年基金资助项目(81702641);江西省自然科学基金杰出青年基金资助项目(2018ACB21042);江西省自然科学基金青年基金资助项目(20171BAB215046);江西省卫生计生委普通科技计划资助项目(20195434)

摘　　要：目的:探讨乳腺癌微环境肿瘤相关成纤维细胞(CAFs)中G蛋白偶联雌激素受体(GPER)活化对成纤维细胞生长因子2(FGF2)的表达调控作用及其诱导的旁分泌对肿瘤细胞生长的影响。方法:免疫荧光法检测单独培养及与ER+乳腺癌MCF-7与ER-乳腺癌MDA-MB-468细胞共培养条件下CAFs和GPER突变型CAFs中GPER的表达定位。用荧光定量PCR法与ELISA法分别检测17-β雌二醇(E2)或GPER特异性激动剂G1处理后单独培养或与乳腺癌细胞共培养的CAFs细胞以及与乳腺癌细胞共培养的GPER突变型CAFs的FGF2mRNA表达和FGF2分泌水平;流式细胞术与CCK-8法检测与CAFs共培养的两种乳腺癌细胞经E2和G1处理后细胞增殖能力的变化,以及FGF2中和抗体的干预作用。结果:单独培养下,GPER定位于CAFs及GPER突变型CAFs的细胞核,与两种乳腺癌细胞共培养后,CAFs细胞中GPER出现明显胞浆转位,而GPER突变型CAFs无此现象。E2和G1处理后,共培养条件下的CAFs中FGF2的mRNA表达及上清液中的FGF2含量均明显增加(均P<0.05),但单独培养的CAFs与共培养条件下的GPER突变型CAFs无上述变化(均P>0.05)。E2和G1处理后,共培养条件下的两种乳腺癌细胞的增殖能力均明显增强(均P<0.05),但该作用均被FGF2中和抗体取消。结论:在乳腺癌微环境下,CAFs中GPER有明显的胞浆转位,这有利于雌激素/GPER/FGF2通路的活化,从而可能促进在乳腺癌的进展。Objective: To investigate the regulatory effect of the activation of G protein-coupled estrogen receptor (GPER) on expression of fibroblast growth factor 2 (FGF2) in cancer-associated fibroblasts (CAFs) in breast cancer and the effect of its induced paracrine on growth of cancer cells. Methods: The expression localizations of GPER in CAFs and CAFs with GPER mutation cultured alone or co-cultured with ER+ breast cancer MCF-7 cells or ER- breast cancer MDA-MB-468 cells were determined by immunofluorescence assays. The mRNA expression and secretion levels of FGF2 in CAFs cultured alone or co-cultured with breast cancer cells and CAFs with GPER mutation co-cultured with breast cancer cells after treatment with 17 β-estradiolum (E2) or GPER specific agonist G1 were detected by real-time qPCR and ELISA, respectively;the changes in proliferative abilities in the two types of breast cancer cells co-cultured with CAFs after treatment with E2 or G1 were examined by flow cytometry and CCK-8 assays, and the interventional effects of FGF2 neutralizing antibody were also observed. Results: In alone culture condition, GPER was located in the nucleus of the CAFs or CAFs with GPER mutation, while evident cytoplasmic translocation of GPER was seen in CAFs but not in CAFs with GPER mutation after co-culture with the both types of breast cancer cells. After treatment with E2 or G1, the expression levels of FGF2 mRNA in CAFs and FGF2 contents in the culture supernatants were significantly increased in co-culture conditions (all P<0.05), but above changes were not observed in CAFs cultured alone and CAFs with GPER mutation under co-culture conditions. The proliferative abilities were significantly enhanced in both types of breast cancer cells in co-culture with CAFs after treatment with E2 or G1 (all P<0.05), but these effects were abolished by adding FGF2 neutralizing antibody. Conclusion: In breast cancer microenvironment, there is cytoplasmic translocation of GPER, which may contribute to the activation of estrogen/GPER/FGF2 pathw

关 键 词：乳腺肿瘤 肿瘤相关成纤维细胞 受体 G-蛋白偶联 成纤维细胞生长因子2 旁分泌细胞细胞间通讯 

分 类 号：R737.9[医药卫生—肿瘤]