C5a激活Akt1-ERK1/2通路促进NSCLC细胞的增殖和迁移  

C5a induces the proliferation and migration of NSCLC cells through activation of Akt1-ERK1/2 pathway

作　　者：何庆玲 赵晨卉[2] 王伟民 葛文 李雅 张婧[1] 王迎伟[1] 邱文[1] HE Qingling;ZHAO Chenhui;WANG Weiming;GE Wen;LI Ya;ZHANG Jing;WANG Yingwei;QIU Wen(Department of Immunology,Nanjing Medical University,Nanjing 211166;Department of Oncology,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学免疫学系,江苏南京211166 [2]南京医科大学第一附属医院肿瘤科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2021年第12期1721-1727,共7页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金(81902878)。

摘　　要：目的:探讨C5a对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞增殖和迁移的影响及其潜在的分子机制。方法:RT-PCR和Western blot检测人正常支气管上皮细胞系BEAS-2B和3种NSCLC细胞系(H1703、PC9、H1299)中C5a受体(C5aR)的表达;CCK-8和划痕实验检测C5a对PC9细胞增殖和迁移的影响;Western blot检查C5a刺激PC9细胞后蛋白激酶Akt1、细胞外信号调节激酶1/2(extracellular signal-regulated kinase,ERK1/2)和蛋白激酶C-α(protein kinase C-α,PKC-α)表达及磷酸化水平;用Akt1抑制剂Perifosine和ERK1/2抑制剂U0126分别处理PC9细胞后再给予C5a刺激,Western blot检测Akt1和ERK1/2的表达和磷酸化及其上下游调控关系,CCK-8和划痕实验检测Perifosine和U0126对细胞增殖和迁移的影响。结果:PC9细胞C5aR的表达水平显著高于其他细胞;C5a可明显促进PC9细胞的增殖和迁移能力;C5a刺激PC9细胞后,可显著增强Akt1和ERK1/2的磷酸化;Akt1和ERK1/2抑制剂均能明显降低C5a刺激PC9细胞后引起的细胞增殖和迁移,且Akt1抑制剂不仅能减弱Akt1的磷酸化,还能减弱ERK1/2的磷酸化,而ERK1/2抑制剂则仅能阻断其自身的磷酸化。结论:C5a刺激NSCLC细胞后可能通过激活Akt1-ERK1/2通路促进细胞的增殖和迁移。Objective:This study aims to investigate the effect of C5a on the proliferation and migration of non-small cell lung cancer(NSCLC)cells and its potential molecular mechanism.Methods:The C5a receptor(C5aR)expression in the human normal bronchial epithelial cell line BEAS-2B and three NSCLC cell lines(H1703,PC9,H1299)was examined by RT-PCR and Western blot(WB)analysis.PC9 cell were stimulated with various concentrations of C5a,and cell proliferation and migration were examined by CCK-8 and scratch test,respectively.The PC9 cells were treated with C5a,and then the expression and phosphorylation of Akt1,ERK1/2 and PKC-αwere examined by WB.PC9 cells were treated with Akt1 inhibitor(Perifosine)and ERK1/2 inhibitor(U0126),respectively,followed by C5a stimulation,and then WB was used to examine the expression and phosphorylation levels of Akt1 and ERK1/2 and their upstream and downstream regulatory relationships.The effects of Perifosine and U0126 on PC9 cell proliferation and migration were determined by CCK-8 and scratch test,respectively.Results:The expression level of C5aR in PC9 cells was obviously higher than that of other cells.C5a could significantly promote the proliferation and migration of PC9 cells.In addition,the phosphorylation levels of both Akt1 and ERK1/2 were markedly enhanced in the PC9 cells induced by C5a,but the protein expression did not show significant change.Furthermore,Akt1 and ERK1/2 inhibitors markedly down-regulated the proliferation and migration of PC9 cells caused by C5a stimulation.Akt1 inhibitors not only attenuated Akt1 phosphorylation,but also attenuated ERK1/2 phosphorylation.ERK1/2 inhibitors only attenuated its own phosphorylation.Conclusion:C5a induces the proliferation and migration of NSCLC cells through activation of Akt1/ERK1/2 pathway.

关 键 词：非小细胞肺癌 C5A AKT1 ERK1/2 增殖 迁移 

分 类 号：R392.12[医药卫生—免疫学]