摘要
目的探讨抗程序性死亡配体1（PD－L1）单抗和表皮生长因子受体酪氨酸激酶抑制剂（EGFR－TKI）对EGFR敏感突变肺癌细胞中细胞膜型和可溶性PD－L1表达以及T细胞功能的影响。方法采用流式细胞术和酶联免疫吸附法（ELISA）检测厄洛替尼干预前后EGFR突变及野生型肺癌细胞株中细胞膜型PD－L1和可溶性PD－L1的表达。以抗PD－L1单抗和（或）厄洛替尼处理肿瘤细胞和T细胞，采用细胞计数盒8（CCK－8）法检测共培养体系中T细胞增殖的变化；采用流式细胞术检测厄洛替尼干预后共培养体系中肿瘤细胞和T细胞中细胞膜型PD－L1的表达变化；采用ELISA法检测γ干扰素（IFN－γ）的水平。结果EGFR敏感突变型细胞株PC9和HCC827中高表达细胞膜型PD－L1，其表达率分别为（78.7±3.1）%和（82.7±2.6）%。经厄洛替尼处理后，PC9和HCC827细胞中细胞膜型PD－L1的表达率分别为（64.7±3.1）%和（73.0±2.6）%，均明显下调（均P〈0.05）；PC9和HCC827细胞培养液上清中可溶性PD－L1的含量分别为（0.680±0.120）ng/ml和（0.903±0.047）ng/ml，均明显降低（均P〈0.05）；而EGFR野生型细胞株A549和H1299中细胞膜型PD－L1和可溶性PD－L1的表达均无明显变化。在EGFR突变型肺癌细胞共培养体系中，厄洛替尼干预可促进T细胞的增殖；联合抗PD－L1单抗后，T细胞的增殖能力更加增强（均P〈0.05）。在EGFR野生型肺癌细胞共培养体系中，厄洛替尼干预对T细胞的增殖无明显影响（均P〉0.05）；联合抗PD－L1单抗后，T细胞的增殖能力亦无明显变化（均P〉0.05）。厄洛替尼干预前后，活化T细胞和HCC827细胞共培养组中IFN－γ的分泌水平分别为（856.0±70.3）pg/ml和（1 697.3±161.0）pg/ml，差异有统计学意义（P〈0.001）；细胞膜型PD－L1的表达率分别为（76.2±0.5）%和（50.9±0.9）%，差异亦有统计学意义（P〈0.001）。厄洛替尼干预并不
Objective To investigate the effects of anti-PD-L1 rnonoclonal antibody and EGFR-TKI on expression of soluble PD-L1 and function of T lymphocytes in EGFR-mutated lung cancer cells.Methods Flow cytometry was used to analyze the expression of membrane PD-LI. ELISA was performed to detect the level of sPD-L1 in the supernatant of cultured EGFR-mutated and wild type lung cancer cells before and after erlotinib treatment. After treated with anti-PD-L1 monoclonal antibody alone and in combination with erlotinib, the proliferation of T lymphocytes in co-culture system was measured using Cell Counting Kit-8 （CCK-8） assay. The expression levels of PD-LI and IFN-~ in tumor cells and T lymphocytes treated with erlotinib in co-culture system were analyzed by flow cytomen7 and ELISA, respectively. Results PD-L1 was highly expressed in EGFR-mutated lung cancer PC9 cells （78.7±3.1） % and HCC827 cells （ 82. 7±2.6） %.After treated with erlotinib, the expression rates of membrane PD-L1 in PC9 and HCC827 cells were down-regulated （ 64.7% ± 3.1% and 73.0% ± 2.6%, respectively） , significantly lower than that in the two cell lines without erlotinib treatment （P〈0.05） , and the expression levels of sPD-LI in the supernatant of PC9 and HCC827 ceils were also down-regulated （0.680±0. 120）ng/ml and （0.903±0.047）ng/ml, respectively, significantly lower than that in the two cell lines without erlofinib treatment （P〈0.01）. However, no significant changes of membrane PD-L1 and sPD-L1 expression were found in EGFR wild type lung cancer cells （H1299 and A549） before and after erlotinib treatment. In the co-culture system composed of T cells and EGFR-mutated lung cancer cells, treatment with erlotinib alone promoted the proliferation of T lymphocytes （P〈0.05） , and combined treatment of anti-PD-L1 monoclonal antibody with erlotinib had a stronger effect （P〈0.05）. In the co-culture system composed of T cells and EGFR wild type cell lines, the proliferation of T cells was not changed after
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2016年第12期886-892,共7页
Chinese Journal of Oncology
基金
国家自然科学基金（81272610）
苏州市科技支撑计划（SS201246）
关键词
肺肿瘤
表皮生长因子受体
程序性死亡配体l
T淋巴细胞
细胞免疫
厄洛替尼
Lung neoplasms
Epidermal growth factor receptor, EGFR
Programmed deathligand 1
T lymphocytes
Cellular immunity
Erlotinib
