Zhu Hong, Liu Dan, Deng Shukai
Department of Respiration, First Affiliated Hospital of Southwest Medical University, Luzhou 646000, China.
Department of Respiration, First Affiliated Hospital of Southwest Medical University, Luzhou 646000, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2021 Oct;37(10):891-896.
Objective To investigate the effect of cisplatin (DDP) on the expression of programmed death 1 ligand 1 (PD-L1) in human lung adenocarcinoma A549 cells and its possible mechanism. Methods Human lung adenocarcinoma A549 cells were cultured in vitro and treated with (0, 0.5, 1, 2, 4, 8) mg/L of DDP for 24, 36, 48 hours. CCK-8 assay was used to detect the cell proliferation inhibition rate and the half maximal inhibitory concentration (IC) was calculated. The optimal inhibition time of 48 hours and its IC were selected for subsequent experiments. A549 cells were then randomized into four groups: blank control group, group with DDP (IC), group with 20 μmol/L of mitogen-activated protein kinase kinase inhibitor PD98059, and group with DDP (IC) combined with 20 μmol/L of PD98059. The cells were cultured for 48 hours. The expression of ERK and p-ERK were detected by Western blotting, and PD-L1 expression was detected by flow cytometry. Results Compared with that in the control group, the expression of PD-L1 in A549 cells treated with DDP was up-regulated in a dose-dependent manner. The optimal time was 48 hours and the IC was 3.586 mg/L. Also compared with that in the control group, the expression of p-ERK and PD-L1 in the DDP treatment group increased, and the expression of p-ERK in the group with PD98059 decreased. The expressions of p-ERK and PD-L1 in the group with DDP combined with PD98059 were lower than those in the group with DDP, but higher than those in the group with PD98059. Conclusion DDP up-regulates the expression of PD-L1 in A549 cells by activating the ERK signaling pathway.
目的 探讨顺铂（DDP）对人肺腺癌A549细胞程序性死亡1配体1（PD-L1）表达的影响及其可能机制。方法 体外培养人肺腺癌A549细胞，分别用（0、0.5、1、2、4、8）mg/L的DDP处理24、36、48小时。采用CCK-8法检测细胞增殖抑制率并计算半数抑制浓度（IC）。选取48小时的最佳抑制时间及其IC用于后续实验。然后将A549细胞随机分为四组：空白对照组、DDP（IC）组、20 μmol/L丝裂原活化蛋白激酶激酶抑制剂PD98059组、DDP（IC）联合20 μmol/L PD98059组。细胞培养48小时。采用蛋白质印迹法检测ERK和p-ERK的表达，采用流式细胞术检测PD-L1表达。结果 与对照组相比，DDP处理的A549细胞中PD-L1的表达呈剂量依赖性上调。最佳时间为48小时，IC为3.586 mg/L。同样与对照组相比，DDP处理组中p-ERK和PD-L1的表达增加，PD98059组中p-ERK的表达降低。DDP联合PD98059组中p-ERK和PD-L1的表达低于DDP组，但高于PD98059组。结论 DDP通过激活ERK信号通路上调A549细胞中PD-L1的表达。
