RNAi沉默肺腺癌A549细胞系PD-L1基因表达对其增殖和凋亡的影响

Knockdown the expression of PD- L1 affects cell proliferation and apoptosis in human lung adenocarcinoma cell line A549

目的:沉默肺癌细胞系A549的程序性死亡配体1(programmed death ligand 1,PD-L1)基因,观察其对A549细胞增殖和凋亡的影响。方法:构建PD-L1 shRNA重组质粒p GPU6/PD-L1,并应用脂质体转染法将其转染入A549细胞。RT-PCR法检测PD-L1基因的表达;Western blotting法检测PD-L1蛋白的表达;MTT法检测p GPU6/PD-L1对A549细胞增殖的影响;AnnexinⅤ-FITC/PI双染法检测p GPU6/PD-L1对A549细胞凋亡的影响。结果:成功将p GPU6/PD-L1转染入A549细胞;RT-PCR结果显示p GPU6/PD-L1使A549细胞PD-L1基因表达水平降低;Western blotting结果显示p GPU6/PD-L1转染A549细胞使PDL1蛋白表达减少;MTT结果显示p GPU6/PD-L1能够抑制A549细胞增殖;AnnexinⅤ-FITC/PI双染法检测结果显示p GPU6/PD-L1能够诱导A549细胞凋亡。结论:p GPU6/PD-L1能够下调肺癌细胞A549 PD-L1的表达,抑制细胞增殖,并诱导细胞凋亡。

Objective：Knockdown the programmed death ligand 1 （PD-L1） in human lung adenocarcinoma cell line A549,to investigate its effects on proliferation and apoptosis of A549 cells.Methods：Construction of PD-L1 shRNA plasmid pGPU6/PD-L1,transfeeted into A.549 cells by liposome.The PD-L1 gene expression was detected by Real-time quantitative PCR,the PD-L1 protein expression of each cell group was detected by Western blot,the proliferation of A549 cells was analyzed bv MTT assay,Annexin Ⅴ-FITC/PI double staining tested the apoptosis of A459 cells bv the flow cytometry.Results：The A549 cells were successfully transfected by pGPU6/PD-L1.The result of Real-time quantitative PCR showed that the PD-L1 gene was down-regulated by pGPU6/PD-L1.The significant down-regulatiou of PD-L1 expression caused by transfection of pGPU6/PD-L1 into A549 cells was detected by Western blotting.The proliferation was inhibited successfully,and apoptosis was induced in A549 cells by pGPU6/PD-L1.Conclusion：The pGPU6/PD-L1 could inhibit the proliferation and promote apoptosis of A549.