G蛋白偶联雌激素受体特异性活化促进骨肉瘤SW1353细胞生长的机制  

Mechanism of G Protein Coupled Estrogen Receptor Specific Activation Promoting the Growth of Osteosarcoma SW1353 Cells

作　　者：邹平安 徐楠 陶志伟[1] 王熙 刘哲[1] 余腾骅 ZOU Ping'an;XU Nan;TAO Zhiwei(Jiangxi Cancer Hospital,Nanchang,330029)

机构地区：[1]江西省肿瘤医院,330029 [2]德安县人民医院,330400

出　　处：《实用癌症杂志》2021年第12期1915-1920,共6页The Practical Journal of Cancer

基　　金：国家自然科学基金地区基金项目(编号:82160565);国家自然科学基金青年基金项目(编号:81702641);江西省自然科学基金杰出青年基金项目(编号:2018ACB21042);江西省卫生计生委普通科技计划项目(编号:20185363)。

摘　　要：目的探讨G蛋白偶联雌激素受体(G protein-coupld estrogen receptor,GPER)特异性活化对骨肉瘤细胞生长的影响。方法生物信息学预测临床骨肉瘤组织及对应正常组织中GPER基因表达差异,并应用Western blot法检测两者GPER蛋白表达情况;荧光定量PCR法与Western blot法筛选人成骨细胞系hFOB1.19及3种骨肉瘤细胞系MG-63、HOS、SW1353中GPER高表达细胞株,并利用细胞免疫荧光法检测SW1353细胞中GPER的表达及定位情况;CCK-8法与流式细胞学检测不同药物处理对骨肉瘤SW1353细胞增殖与细胞周期的影响,并采用Western blot法检测细胞周期蛋白cyclinA与cyclinD1的表达情况。结果GPER在肿瘤组织/细胞中呈现高表达(P<0.05),其中骨肉瘤SW1353细胞中GPER表达量最高(P<0.05),其主要表达于细胞核与细胞质中。100 nmol/L药物浓度的GPER特异性激动剂G1可显著促进骨肉瘤SW1353细胞生长及细胞周期进展。与对照组相比,G1处理组中DNA合成期(S期)细胞比例明显升高,同时伴有细胞周期蛋白cyclinA与cyclinD1的表达增加(P<0.05)。有趣的是,GPER特异性拮抗剂G15显著抑制G1所诱发的上述改变(P<0.05)。结论GPER特异性活化能明显上调细胞周期蛋白cyclinA与cyclinD1的表达,促进骨肉瘤SW1353细胞周期进展,从而促进骨肉瘤细胞异常生长。Objective To investigate the effect of G protein-coupled estrogen receptor(GPER)-specific activation on the growth of osteosarcoma cells.Methods Bioinformatics predicts GPER gene expression differences in clinical osteosarcoma tissues and the corresponding normal tissues,and the protein expression of GPER was determined by western blot.GPER highly expressed cell lines were screened in the human osteoblastic cell line hFOB1.19 and 3 osteosarcoma MG-63,HOS and SW1353 cell lines by quantitative PCR and western blot,the expression and localization of GPER in SW1353 cells were determined by immunofluorescence.CCK-8 and flow cytometry were performed to detect the effect of different drug trea3tment on osteosarcoma SW1353 cell proliferation and cell cycle,the expression of cyclinA and cyclinD1 was also determined by the western blot.Results GPER was highly expressed in tumor tissues/cells(P<0.05),with the highest expression in osteossarcoma SW1353 cells(P<0.05),which was mainly expressed in the nucleus and cytoplasm.The GPER-specific agonist G1 at 100 nmol/L drug concentration significantly promoted osteosarcoma SW1353 cell growth and cell cycle progression.The proportion of G1-induced DNA synthesis phase(S phase)cells was significantly increased along with enhanced cyclinA and cyclinD1 expression when compared with control groups(P<0.05).Interestingly,the GPER-specific antagonist G15 significantly inhibited the aforementioned changes wihch induced by G1(P<0.05).Conclusion GPER-specific activation can significantly upregulate cyclinA and cyclinD1 expression and promote the development of osteosarcoma SW1353 cell cycle progression,promoting abnormal growth of osteosarcoma cell.

关 键 词：GPER 骨肉瘤 肿瘤生长 

分 类 号：R738.7[医药卫生—肿瘤]