论文部分内容阅读
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: To explore the expression of PD-L1 in lung cancer cells and its effect on cell migration. Methods: Flow cytometry was used to detect the expression of PD-L1 on the surface of lung cancer cell lines with different pathological types. The cell lines with high, medium and low expression were selected, and then induced by adding cytokines to lung cancer cell lines. The expression of PD-L1 in lung cancer cell lines was detected by flow cytometry. The expression of PD-L1 in lung cancer cell lines with moderate and high expression of PD-L1 was down-regulated by siRNA interference. The transwell assay was used to detect the changes of cell migration. Results: The expression of PD-L1 on the surface of HCC827, H1299, A549 (adenocarcinoma), H226, H520 (squamous cell carcinoma), H460 (large cell carcinoma) and H446 (small cell carcinoma) were HCC827 and H460 H1299, H226, H446, H520 (middle expression), A549 (low expression). The adenocarcinoma cell lines H1299 and A549 with transwell migration ability were selected and cultured with the addition of cytokines IL-4, IL-6, IL-13, TGF-β-1 and IFN- -γ could up-regulate the expression of PD-L1 in lung cancer cell lines. The results of transwell assay showed that the migration ability of cells was significantly enhanced compared with the control group. The PD-L1 expression of PD-L1 cells was significantly down-regulated after si-PD-L1 interfered the expression of PD-L1 in lung cancer cell lines. The migration ability of PD-L1 cells was significantly weaker than that of the control cells. After up-regulation, the ability of cell migration was enhanced. After siRNA interference, the expression of PD-L1 was down-regulated, and the cell migration ability was significantly weakened. Conclusion: The expression of PD-L1 can promote the migration of lung cancer cells and may be involved in lung cancer metastasis.