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第41卷第12期 南京医科大学学报(自然科学版)
2021年12月 Journal of Nanjing Medical University(Natural Sciences) ·1721 ·
·基础医学·
C5a激活Akt1⁃ERK1/2通路促进NSCLC细胞的增殖和迁移
何庆玲 ,赵晨卉 ,王伟民 ,葛 文 ,李 雅 ,张 婧 ,王迎伟 ,邱 文 1*
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1 南京医科大学免疫学系,江苏 南京 211166;南京医科大学第一附属医院肿瘤科,江苏 南京 210029
[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。
[关键词] 非小细胞肺癌;C5a;Akt1;ERK1/2;增殖;迁移
[中图分类号] R392.12 [文献标志码] A [文章编号] 1007⁃4368(2021)12⁃1721⁃07
doi:10.7655/NYDXBNS20211202
C5a induces the proliferation and migration of NSCLC cells through activation of Akt1 ⁃
ERK1/2 pathway
1 2 1 1 1 1 1 1*
HE Qingling ,ZHAO Chenhui ,WANG Weiming ,GE Wen ,LI Ya ,ZHANG Jing ,WANG Yingwei ,QIU Wen
1 Department of Immunology,Nanjing Medical University,Nanjing 211166;Department of Oncology,the First
2
Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China
[Abstract] 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⁃α were 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.
[Key words] NSCLC;C5a;Akt1;ERK1/2;proliferation;migration
[J Nanjing Med Univ,2021,41(12):1721⁃1727]
[基金项目] 国家自然科学基金(81902878)
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通信作者(Corresponding author),E⁃mail:qiuwen@njmu.edu.cn