下调PD-L1表达对吉非替尼耐药性非小细胞肺癌细胞的影响  

Effects of down-regulating PD-L1 expression on gefitinib-resistant non-small cell lung cancer cells

作　　者：姬颖华[1] 杨晓煜[2] 孟祥丽[1] 王瑾 路平[1] Ji Yinghua;Yang Xiaoyu;Meng Xiangli;Wang Jin;Lu Ping(Depatment of Oncology,The First Affiliated Hospital of Xinxiang Medical University;Department of Pathology,School of Basic Medicine,Xinxiang Medical College)

机构地区：[1]新乡医学院第一附属医院肿瘤科,卫辉453100 [2]新乡医学院基础医学院病理科,新乡453003

出　　处：《重庆医科大学学报》2022年第2期156-161,共6页Journal of Chongqing Medical University

基　　金：河南省医学科技攻关计划资助项目(编号:2018020369)。

摘　　要：目的:研究下调程序性细胞死亡配体1(programmed cell death ligand 1,PD-L1)表达对吉非替尼耐药性(gefitinibresistant,GR)非小细胞肺癌细胞的作用及机制。方法:通过RT-qPCR和Western blot检测各非小细胞肺癌细胞程序性细胞死亡1(programmed cell death 1,PD1)和PD-L1 mRNA和其蛋白表达水平;获得GR细胞H1703/GR,CCK-8检测细胞活性,Western blot检测PD1和PD-L1蛋白表达;转染sh-PD-L1进入H1703/GR细胞,CCK-8检测细胞活性,流式细胞术检测细胞凋亡,Western blot检测PI3K/Akt/mTOR通路相关蛋白的表达;加入PI3K/Akt/mTOR通路激活剂IGF-1,CCK-8检测细胞活性,流式细胞术检测细胞凋亡。结果:与人永生化肺上皮细胞株16HBE相比,非小细胞肺癌细胞株GLC82、A549、PC9、SK-MES-1、H1703、L78、H460、H661中PD1和PD-L1 mRNA及蛋白表达均上调(P<0.05、P<0.01),选取H1703细胞进行后续实验;与H1703细胞相比,H1703/GR细胞活性升高,PD1和PD-L1蛋白表达均上调(P<0.01);与GR组相比,GR+sh-PD-L1组H1703/GR细胞活性明显降低,细胞凋亡率明显增加,p-Akt/Akt和p-mTOR/mTOR蛋白表达明显下调(P<0.01);与GR+sh-PD-L1组相比,GR+sh-PD-L1+IGF-1组H1703/GR细胞活性明显升高,细胞凋亡率明显减少(P<0.01)。结论:下调PD-L1表达可能通过抑制PI3K/Akt/mTOR通路激活,增强非小细胞肺癌对吉非替尼的敏感性。Objective:To investigate the effect and mechanism of down-regulation of programmed cell death ligand 1(PD-L1 expression on gefitinib-resistant(GR)non-small cell lung cancer cells. Methods:The expression levels of programmed cell death 1(PD1)and PD-L1 mRNA and protein in non-small cell lung cancer cells were detected by RT-qPCR and Western blot. GR cells H1703/GR were obtained. Cell activity was detected by CCK-8. Western blot was used to detect PD1 and PD-L1 protein expression.Sh-PD-L1 was transfected into H1703/GR cells,cell activity was detected by CCK-8,apoptosis was detected by flow cytometry,and PI3 K/Akt/mTOR pathway-related protein expression was detected by Western blot. PI3 K/Akt/mTOR pathway activator IGF-1 was added,cell activity was detected by CCK-8 and apoptosis was detected by flow cytometry. Results:Compared with human immortalized lung epithelial cell line 16 HBE,the expressions of PD1 and PD-L1 mRNA and protein in non-small cell lung cancer cell lines GLC82,A549,PC9,SK-MES-1,H1703,L78,H460,and H661 were all up-regulated(P<0.05,P<0.01),and H1703 cells were selected for subsequent experiments. Compared with H1703 cells,H1703/GR cells increased activity,and both PD1 and PD-L1 protein expressions were up-regulated(P<0.01). Compared with GR group,GR+sh-PD-L1 group significantly reduced H1703/GR cell viability,increased apoptotic rate and decreased p-Akt/Akt and p-mTOR/mTOR protein expression(P<0.01). Compared with GR+sh-PDL1 group,the activity of H1703/GR cells in GR +sh-PD-L1 +IGF-1 group was significantly increased and the apoptosis rate was significantly reduced(P <0.01). Conclusion:Down-regulating PD-L1 expression enhances the sensitivity of non-small cell lung cancer to gefitinib by inhibiting activation of the PI3 K/Akt/m TOR pathway.

关 键 词：非小细胞肺癌 吉非替尼 PD1 PD-L1 

分 类 号：R734.2[医药卫生—肿瘤]