血浆外泌体淋巴细胞白血病缺失基因1水平对表皮生长因子受体/间变性淋巴瘤激酶野生型进展期非小细胞肺癌患者免疫治疗反应的预测价值

Predictive value of plasma exosomal deleted in lymphocytic leukemia 1 level in immunotherapy response of epidermal growth factor receptor/anaplastic lymphoma kinase wild type advanced non-small cell lung cancer patients

目的 探讨血浆外泌体淋巴细胞白血病缺失基因1(exoDLEU1)在预测表皮生长因子受体(EGFR)/间变性淋巴瘤激酶(ALK)野生型进展期非小细胞肺癌(NSCLC)患者免疫治疗反应中的临床意义。方法 收集2017年6月至2019年2月于浙江省湖州市第一人民医院接受免疫治疗的91例EGFR/ALK野生型进展期NSCLC患者的血液样本,根据免疫治疗后的反应将其分为反应组(53例)和无反应组(38例)。提取两组治疗前血浆外泌体,通过实时荧光定量PCR检测两组exoDLEU1表达,采用logistic回归分析EGFR/ALK野生型进展期NSCLC患者免疫治疗反应情况的影响因素,绘制受试者操作特征(ROC)曲线评估血浆exoDLEU1水平对患者免疫治疗反应情况的预测价值。结果 透射电镜下可见囊泡具有圆形或椭圆形膜,直径为30~150 nm,原始浓度为6.6×10^(10)个粒子/ml,流式细胞仪检测及蛋白质印迹法结果显示,提取物含有外泌体标志物CD63、CD9、CD81、热休克蛋白70。实时荧光定量PCR结果显示,反应组治疗前血浆exoDLEU1水平低于无反应组,差异有统计学意义(P<0.05)。影响因素分析结果显示,血浆exoDLEU1表达水平是EGFR/ALK野生型进展期NSCLC患者免疫治疗无反应的独立影响因素(OR=6.119,95%CI:2.768~13.526,P<0.05)。ROC曲线分析显示,血浆exoDLEU1表达水平预测EGFR/ALK野生型进展期NSCLC患者免疫治疗反应情况的曲线下面积为0.900 (P<0.05)。结论 EGFR/ALK野生型进展期NSCLC患者免疫治疗前的血浆exoDLEU1水平可作为预测其治疗反应的指标,有助于在临床中筛选出免疫治疗更易获益的患者。

Objective To investigate the clinical significance of plasma exosomal deleted in lymphocytic leukemia 1(exo-DLEU1)in predicting immunotherapy response of epidermal growth factor receptor(EGFR)/anaplastic lymphoma kinase(ALK)wild type advanced non-small cell lung cancer(NSCLC)patients.Methods The blood samples of 91 patients with EGFR/ALK wild type advanced NSCLC who received immunotherapy in the First People’s Hospital of Huzhou,Zhejiang Province from June 2017 to February 2019 were collected and divided into response group(53 cases)and non-response group(38 cases)according to the response after immunotherapy.Plasma exosomes were extracted from the two groups before treatment,and exoDLEU1 expression was detected by real-time fluorescent quantitative PCR.The influencing factors of immunotherapy response in EGFR/ALK wild type advanced NSCLC patients were analyzed by logistic regression.Receiver operator characteristic(ROC)curve was plotted to evaluate the predictive value of plasma exoDLEU1 level in immunotherapy response.Results Under transmission electron microscopy,the vesicles had circular or elliptical membranes with diameters ranging from 30 to 150 nm,and the original concentration was 6.6×10^(10) particles/ml.Flow cytometry and Western blot showed that the extracts contained exosome markers CD63,CD9,CD81,and heat shock protein 70.Real-time fluorescence quantitative PCR results showed that the plasma exoDLEU1 level in the response group before treatment was lower than that in the non-response group,and the difference was statistically significant(P<0.05).Analysis of influencing factors showed that plasma exoDLEU1 expression level was an independent influencing factor for immunotherapy non-response in EGFR/ALK wild type advanced NSCLC patients(OR=6.119,95%CI:2.768-13.526,P<0.05).ROC curve analysis showed that plasma exoDLEU1 expression level predicted immunotherapy response of EGFR/ALK wild type advanced NSCLC patients with an area under the curve of 0.900(P<0.05).Conclusion The plasma exoDLEU1 level of EGFR/ALK wild type advanced NSCLC patients before immunotherapy can be used as an indicator to predict their treatment response,and help to screen out patients who are more likely to benefit from immunotherapy in the clinic.