PD-L1对肺癌细胞迁移能力的影响

摘要

目的:探索PD-L1在肺癌细胞中的表达及其对细胞迁移能力的影响.方法:采用流式细胞术检测不同病理类型肺癌细胞株表面PD-L1分子表达水平,分别选择高、中、低表达的细胞株,后添加细胞因子诱导肺癌细胞株,采用免疫印迹术筛选出能够提高肺癌细胞株PD-L1分子表达水平的细胞因子;采用siRNA干扰技术,下调中、高表达PD-L1的肺癌细胞株中PD-L1的分子表达水平,transwell实验检测细胞迁移能力改变.结果:PD-L1在细胞株HCC827、H1299、A549(腺癌)、H226、H520(鳞癌)、H460(大细胞癌)、H446(小细胞癌)表面的表达水平分别为HCC827、H460(高表达),H1299、H226、H446、H520(中表达),A549(低表达).选择具有transwell迁移能力的腺癌细胞株H1299、A549,分别添加细胞因子IL-4、IL-6、IL-13、TGF-β-1、IFN-γ培养,筛选出TGF-β-1、IFN-γ两种因子能够上调肺癌细胞株PD-L1的表达水平,transwell实验结果显示细胞迁移能力较对照组显著增强.si-PD-L1干扰肺癌细胞株PD-L1表达后,PD-L1的表达水平明显下调,细胞迁移能力较对照组显著减弱;TGF-β-1、IFN-γ诱导细胞株PD-L1表达水平上调后能够引起细胞迁移能力增强,经siRNA干扰后,PD-L1表达水平下调,细胞迁移能力随之显著减弱.结论:PD-L1的表达能促进肺癌细胞的迁移,可能参与肺癌转移过程.%Objective:Screening of the pathological types of lung cancer cell line PD-L1 molecular expression levels,through the up and down PD-L1 expression level,to explore its effect on the migration of lung cancer cells.Methods:By flow cytometry in different pathological types of lung cancer cell line surface of PD-L1 expression levels were detected.Selecting high,medium and low expression cell line,after adding cytokines co cultured lung cancer cell lines.Western blot were used to select cytokines witch can improve the lung cancer cell line of PD-L1 expression levels.Using small interfering RNA(si-RNA)intervention,high expression molecules PD-L1 lung cancer cell line of PD-L1 expression levels and transwell experiments was used to detect the cell migration ability to change.Results:PD-L1 expression level of HCC827(adenocarcinoma)and H1299(adenocarcinoma),A549(adenocarcinoma),H226(squamous cell carcinoma),H520(squamous cell carcinoma),H460(large cell lung cancer),H446(small cell carcinoma)surface were high in HCC827,H460 and medium in H1299,H226,H520,H446,low A549 in.Choose to have the ability to migrate adenocarcinoma cell line H1299 and A549,were added to IL-4,IL-6,IL-13,TGF-β-1,IFN-γ.Selected TGF-β-1,IFN-γ two factors which can upregulate PD-L1 expression levels and cell migration ability than that of the control group was significantly increased.With si-PD-L1 interference of lung cancer cell line PD-L1 expression,PD-L1 expression was significantly down regulated,the cell migration ability was significantly decreased compared with the control group.TGF-β-1 and IFN-γ induced cell line PD-L1 expression level up regulation,after siRNA interference,PD-L1 expression level was down,cell migration ability decreased significantly.Conclusion:The expression level of PD-L1 was correlated with the migration of lung cancer cell lines.