桥接整合因子1通过c-MYC途径抑制非小细胞肺癌A549细胞中PD-L1的表达  

Bridging intergrator-1 inhibits the expression of PD-L1 through c-MYC pathway in non-small cell lung cancer A549 cells

作　　者：李欢 王梦杰 张翔宇[1] 段玉青[1] 贾云泷[1] 刘欣燕[2] 刘丽华[1] LI Huan;WANG Mengjie;ZHANG Xiangyu;DUAN Yuqing;JIA Yunlong;LIU Xinyan;LIU Lihua(Department of Tumor Immunotherapy,Fourth Hospital of Hebei Medical University,Shijiazhuang 050035,Hebei,China;Department of Oncology,Hebei Chest Hospital,Shijiazhuang 050035,Hebei,China)

机构地区：[1]河北医科大学第四医院肿瘤免疫治疗科,河北石家庄050035 [2]河北省胸科医院肿瘤科,河北石家庄050035

出　　处：《中国肿瘤生物治疗杂志》2018年第8期762-766,共5页Chinese Journal of Cancer Biotherapy

基　　金：国家自然科学基金资助项目(No.81201607);河北杰出青年基金资助项目(No.H2014206320);河北人才培养项目资助(No.A201401040)~~

摘　　要：目的:研究人非小细胞肺癌(non-small cell lung cancer,NSCLC)A549细胞中桥接整合因子-1(bridging intergrator-1,BIN1)对程序性死亡受体-配体1(programmed death-ligand 1,PD-L1)表达的影响及其机制。方法:采用q RT-PCR和Western blotting方法检测A549细胞和正常人胚肺成纤维细胞2BS中BIN1与PD-L1基因和蛋白表达情况,通过基因转染技术、利用阳离子脂质体将含有人全长BIN1基因序列的真核表达质粒CMV-MCS-GFP-SV40-Neomycin-BIN1转染到A549细胞中(BIN1+组),构建过表达BIN1的细胞株,采用RNA干扰技术将干扰骨髓细胞瘤病毒癌基因(cellular-myelocytomatosis viral oncogene,c-MYC)的c-MYC-si RNA转染到A549细胞中(c-MYC-si RNA组)以敲低c-MYC基因表达,通过q RT-PCR和Western blotting方法验证转染效果及过表达BIN1基因或敲低c-MYC基因对A549细胞中c-MYC和PD-L1表达的影响。结果:与2BS细胞相比,A549细胞中BIN1基因和蛋白均呈低表达状态,而PD-L1呈高表达状态(均P<0.05)。将携带BIN1基因的真核表达质粒转染到A549细胞后,BIN1基因和蛋白表达水平较对照组显著升高(P<0.05),而PD-L1表达显著降低(P<0.05)。c-MYC-si RNA转染到A549细胞后,细胞内c-MYC表达显著降低(P<0.01),PD-L1表达明显下调(P<0.01)。结论:BIN1过表达可以通过失活c-MYC通路降低PDL1表达,从而抑制A549细胞的免疫逃逸。Objective： To investigate the effect and mechanism of bridging intergrator-1（BIN1） on expression of programmed death-ligand 1（PD-L1） in non-small cell lung cancer（NSCLC） A549 cells. Methods： Quantitative real-time polymerase chain reaction（q RTPCR） and Western blotting were used to detect the m RNA and protein expression of BIN1 and PD-L1 in A549 cells and normal human embryo lung fibroblast 2 BS cells, respectively. Eukaryotic expression plasmid CMV-MCS-GFP-SV40-Neomycin-BIN1 containing human full length BIN1 gene sequence was transfected into A549 cells via cationic liposomes by using gene transfection technology（as BIN1＋group）; c-MYC-si RNA was used to knockdown the expression of c-MYC in A549 cells through RNA interference technique（as cMYC-si RNA group）. The transfection efficiencies were verified by q RT-PCR and Western blotting, the effects of BIN1 over-expression and c-MYC knock-down on the expression of c-MYC and PD-L1 in A549 cells were detected as well. Results： Comparing with 2 BS cells, the expression of BIN1 was down-regulated in A549 cells at both m RNA and protein levels, while the expression of PD-L1 was up-regulated（all P〈0.05）. The expression of BIN1 was increased at both m RNA and protein level in BIN1＋group, while the expression of PD-L1 was decreased significantly after B1 N1 transfection（all P〈0.05）. After transfection of c-MYC-si RNA into A549 cells, the expression of c-MYC and PD-L1 in c-MYC-si RNA group was down-regulated significantly（all P〈0.01）. Conclusion： Over-expression of BIN1 could reduce the expression of PD-L1 by inactivating the c-MYC pathway, thereby inhibiting the immune escape of A549 cells.

关 键 词：非小细胞肺癌 A549细胞 桥接整合因子-1 骨髓细胞瘤病毒癌基因 程序性死亡受体-配体1 免疫逃逸 

分 类 号：R730.2[医药卫生—肿瘤] R734.2[医药卫生—临床医学]