晚期结直肠癌西妥昔单抗治疗前后循环肿瘤DNA、循环B细胞特异性莫洛尼白血病病毒插入位点1 mRNA和微小RNA-21水平变化

Changes of circulating tumor DNA, circulating B cell-specific Moloney leukemia virus insertion site 1 mRNA and microRNA-21 levels before and after cetuximab treatment in advanced colorectal cancer

目的探讨晚期结直肠癌表皮生长因子受体(EGFR)单克隆抗体治疗前后循环肿瘤DNA(ctDNA)、循环B细胞特异性莫洛尼白血病病毒插入位点1 mRNA(Bmi-1mRNA)、微小RNA-21(miR-21)变化及与治疗反应性的关联性。方法回顾性分析2019年3月至2021年3月烟台毓璜顶医院98例晚期结直肠癌患者的临床资料。西妥昔单抗治疗后完全缓解4例,部分缓解26例,疾病稳定39例,疾病进展29例。将完全缓解和部分缓解作为缓解组(30例),疾病稳定和疾病进展作为未缓解组(68例)。治疗前和治疗2个周期后采用高通量测序方法检测血浆ctDNA水平,采用实时荧光定量聚合酶链反应法检测Bmi-1mRNA和miR-21水平。采用Spearman相关性分析治疗2个周期后ctDNA、Bmi-1mRNA和miR-21与治疗反应性的关系;采用多因素Logistic回归分析影响治疗反应性的独立危险因素;绘制受试者工作特征(ROC)曲线,评估治疗2个周期后ctDNA、Bmi-1mRNA和miR-21预测缓解的价值。结果两组治疗前ctDNA、Bmi-1mRNA和miR-21比较差异无统计学意义(P>0.05);缓解组治疗2个周期后ctDNA、Bmi-1mRNA和miR-21明显低于未缓解组[(10.03±3.32)μg/L比(15.33±5.14)μg/L、0.13±0.04比0.19±0.05和0.81±0.26比1.08±0.24],差异有统计学意义(P<0.01)。Spearman相关性分析结果显示,ctDNA、Bmi-1mRNA和miR-21与治疗反应性呈负相关(r=-0.500、-0.506和-0.531,P<0.01)。多因素Logistic回归分析结果显示,控制远处转移器官数量和临床分期后,ctDNA、Bmi-1mRNA和miR-21仍是影响晚期结直肠癌患者治疗反应性的独立危险因素(OR=3.342、2.725和1.838,95%CI 3.116~3.584、2.647~2.805和1.768~1.911,P<0.01)。ROC曲线分析结果显示,治疗2个周期后ctDNA、Bmi-1mRNA联合miR-21预测治疗反应性的曲线下面积最大为0.922。结论晚期结直肠癌患者EGFR单克隆抗体治疗前后ctDNA、Bmi-1mRNA和miR-21变化与治疗反应性有关,联合检测有利于筛查EGFR靶向治疗的敏感患者,并能为寻找晚期结直肠癌分子干预新靶点提供参考。

Objective To investigate the changes of circulating tumor DNA(ctDNA),circulating B cell-specific Moloney leukemia virus insertion site 1 mRNA(Bmi-1 mRNA)and microRNA-21(miR-21)before and after treatment with epidermal growth factor receptor(EGFR)monoclonal antibody in advanced colorectal cancer,and analyze their association with treatment response.Methods The clinical data of 98 patients with advanced colorectal cancer from March 2019 to March 2021 in Yantai Yuhuangding Hospital were retrospectively analyzed.After treatment with cetuximab,complete remission was in 4 cases,partial remission in 26 cases,stable disease in 39 cases,and progressive disease in 29 cases.The patients with complete remission and partial remission were classified as remission group(30 cases),the stable disease and progressive disease were classified as non-remission group(68 cases).Before treatment and after 2 cycles of treatment,the plasma level of ctDNA was detected by high-throughput sequencing;the levels of Bmi-1mRNA and miR-21 were detected by real-time fluorescence quantitative polymerase chain reaction.Spearman correlation was used to analyze the relationship between ctDNA,Bmi-1mRNA,miR-21 and treatment responsiveness after 2 cycles of treatment;multivariate Logistic regression was used to analyze the independent risk factors affecting treatment responsiveness;receiver operating characteristic(ROC)curve was drawn to evaluate the value of ctDNA,Bmi-1mRNA and miR-21 in predicting remission after 2 cycles of treatment.Results There were no significant differences in ctDNA,Bmi-1mRNA and miR-21 before treatment between 2 groups(P>0.05);the ctDNA,Bmi-1mRNA and miR-21 after 2 cycles of treatment in remission group were significantly lower than those in non-remission group:(10.03±3.32)μg/L vs.(15.33±5.14)μg/L,0.13±0.04 vs.0.19±0.05 and 0.81±0.26 vs.1.08±0.24,and there were statistical differences(P<0.01).Spearman correlation analysis result showed that ctDNA,Bmi-1mRNA and miR-21 were negatively correlated with treatment response(r=-0.500,-0.506 and-0.531;P<0.01).Multivariate Logistic regression analysis result showed that,after controlling for the number of distant metastatic organs and clinical stage,ctDNA,Bmi-1mRNA and miR-21 were still independent risk factors for treatment response in patients with advanced colorectal cancer(OR=3.342,2.725 and 1.838;95%CI 3.116 to 3.584,2.647 to 2.805 and 1.768 to 1.911;P<0.01).ROC curve analysis result showed that the area under the curve(AUC)of ctDNA,Bmi-1mRNA combined with miR-21 after 2 cycles of treatment to predict the treatment response was the largest with 0.922.Conclusions The changes of ctDNA,Bmi-1mRNA and miR-21 in patients with advanced colorectal cancer before and after treatment with EGFR monoclonal antibody are related to the treatment response.Combined detection is helpful for screening patients sensitive to EGFR-targeted therapy,and can provide reference for new targets of molecular intervention.