miR-200通过靶向PD-L1抑制非小细胞肺癌A549细胞增殖、迁移和侵袭的研究  

Study of MiR-200 inhibiting proliferation,migration and invasion of non-small cell lung cancer A549 cells by targeting PD-L1

作　　者：艾孜子·阿不来提 艾力江·多力坤 陈康[1] AIZIZI·Abulaiti;AILIJIANG·Duolikun;CHEN Kang(Department of Thoracic Surgery,Xinjiang Uygur Autonomous Region People′s Hospital,Urumqi,Xinjiang 830001,China)

机构地区：[1]新疆维吾尔自治区人民医院胸外科,新疆乌鲁木齐830001

出　　处：《临床肺科杂志》2021年第10期1551-1557,共7页Journal of Clinical Pulmonary Medicine

摘　　要：目的探讨miR-200对非小细胞肺癌A549细胞的增殖、迁移和侵袭的影响及其与程序性死亡受体配体-1(Programmed death-ligand 1,PD-L1)的靶向关系。方法选取非小细胞肺癌细胞株A549,正常肺成纤维细胞HLF-1为研究对象,根据A549细胞转染物质的不同,分为NC组(未进行任何处理的A549细胞)、miR-NC组(转染miR-NC)、miR-200组(转染miR-200-mimic)、si-NC组(转染si-NC)、si-PD-L1组(转染si-PD-L1)、miR-200+pEGFP组(转染miR-200-mimic+pEGFP)及miR-200+pEGFP-PD-L1组(转染miR-200-mimic+pEGFP-PD-L1)。采用实时荧光定量PCR法检测细胞中miR-200表达水平;MTT检测细胞增殖能力;Transwell小室实验检测细胞迁移、侵袭能力;Western blot检测细胞PD-L1表达水平;荧光素酶实验验证miR-200与PD-L1的靶向关系。结果与HLF-1相比,A549细胞胞miR-200表达水平降低,PD-L1基因、蛋白表达升高(P<0.05)。48和72 h时,miR-200组细胞活力、迁移和侵袭数量明显低于NC组、miR-NC组(P<0.05)。48和72 h时,si-PD-L1组细胞活力、迁移和侵袭数量明显低于NC组、si-NC组(P<0.05)。miR-200+pcDNA-PD-L1组细胞48、72 h时细胞活力、细胞迁移和侵袭数量高于miR-200+pcDNA组、miR-200组(P<0.05)。与对照组相比,A549细胞PD-L1野生型质粒组荧光素酶活性明显降低(P<0.05),PD-L1突变型质粒组荧光素酶活性无明显变化(P>0.05)。结论miR-200可负性调控PD-L1抑制非小细胞肺癌A549细胞增殖、迁移和侵袭。Objective To investigate the effect of miR-200 on the proliferation,migration and invasion of non-small cell lung cancer A549 cells and its targeting relationship with programmed death ligand-1(PD-L1).Methods Non-small cell lung cancer cell lines A549 and normal lung fibroblast HLF-1 were selected.According to the different transfection substances,they were divided into the NC group(A549 cells),the miR-NC group(miR-NC),the miR-200 group(miR-200-mimic),the si-NC group A(transfected with si-NC),the si-PD-L1 group(transfected with si-PD-L1),the miR-200+pEGFP group(transfected with miR-200-mimic+pEGFP)and the miR-200+pEGFP-PD-L1 group(transfected with miR-200-mimic+pEGFP-PD-L1).Real time fluorescent quantitative PCR was used to detect the expression of miR-200.MTT was used to detect cell proliferation.Transwell chamber test was used to detect cell migration and invasion.Western blot was used to detect the expression level of PD-L1.Luciferase assay was used to verify the relationship between miR-200 and PD-L1.Results Compared with HLF-1,the expression of miR-200 was decreased,and PD-L1 gene and protein expression were increased in non-small cell lung cancer cell line A549(P<0.05).At 48 and 72 h,the cell viability,migration and invasion of the miR-200 group were significantly lower than those of the NC group and the miR-NC group(P<0.05).At 48h and 72 h,the cell viability,migration and invasion in the si-PD-L1 group were significantly lower than those in the NC group and the Si-NC group(P<0.05).The cell viability,migration and invasion of the miR-200+pcDNA-PD-L1 group were higher than those of the miR-200+pcDNA group and the miR-200 group at 48 and 72 h(P<0.05).The luciferase reporter assay showed that miR-200 could reduce the activity of luciferase in PD-L1 wild-type vectors(P<0.05),but there was no statistically significant difference in PD-L1 mutant vectors(P>0.05).Conclusion miR-200 can negatively regulate PD-L1 and inhibit the proliferation,migration and invasion of non-small cell lung cancer A549 cells.

关 键 词：miR-200 PD-L1 非小细胞肺癌 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]