机构地区:[1]邢台市第三医院胸外科,河北邢台054000 [2]冀中能源邢台矿业集团有限责任公司总医院医务科,河北邢台054000 [3]邯郸市第二医院心内二科,河北邯郸056001
出 处:《临床和实验医学杂志》2021年第9期948-953,共6页Journal of Clinical and Experimental Medicine
基 金:邢台市科技计划项目(编号:2017ZC086);河北省卫生厅科技计划项目(编号:20181662)。
摘 要:目的分析肝激酶B1(LKB1)-腺苷酸活化蛋白激酶(AMPK)-雷帕霉素靶蛋白(mTOR)通路对患者胸腔积液性质的诊断价值。方法前瞻性选取2018年6月至2020年6月期间邢台市第三医院收治的168例胸腔积液肿瘤患者为研究对象,其中92例恶性胸水患者作为恶性组,其余76例良性胸水患者作为良性组。抽提2组患者胸腔积液内细胞总RNA和总蛋白,免疫组织化学法检测并比较LKB1、AMPK、mTOR表达,蛋白质印迹法检测并比较AMPKα、p-AMPKα、LKB1、mTOR、p-mTOR蛋白表达,实时荧光定量PCR检测并比较LKB1、AMPKα和mTOR mRNA表达,并分析LKB1、AMPK、mTOR在胸腔积液中的诊断效果。结果免疫组织化学法显示,2组胸腔积液内LKB1、AMPK、mTOR阳性染色均呈现棕黄色,主要于细胞浆内表达。恶性组的LKB1、AMPK的阳性率均低于良性组,mTOR阳性率高于良性组,差异均有统计学意义(P<0.05)。免疫蛋白印迹实验显示,2组胸腔积液内p-AMPKα蛋白相对表达量比较,差异无统计学意义(P>0.05);恶性组的AMPKα、p-mTOR蛋白相对表达量高于良性组,LKB1、mTOR蛋白相对表达量低于良性组,差异均有统计学意义(P<0.05)。实时荧光定量PCR显示,恶性组的LKB1、AMPKαmRNA表达均低于良性组,mTOR mRNA表达高于良性组,差异均有统计学意义(P<0.05)。接收者工作特征(ROC)曲线分析显示:LKB1、AMPK、mTOR等指标具有较高的对患者胸腔积液性质的诊断价值,曲线下面积(AUC)分别为0.861、0.831、0.852,95%CI分别为0.774~0.958、0.686~0.992、0.586~1.000;联合应用对胸腔积液性质的诊断AUC为0.918,95%CI为0.790~1.000,其诊断效能比单一指标有明显提高。结论LKB1-AMPK-mTOR通路对肿瘤患者胸腔积液良、恶性具有较佳的诊断价值。Objective To analyze the diagnostic value of liver kinase B1(LKB1)-adenylate activated protein kinase(AMPK)-target rapamycin(mTOR)pathway in the nature of pleural effusion in patients.Methods Using prospective analysis,a prospective analysis method was adopted to select 168 patients with pleural effusion tumor admitted to the Third Hospital of Xingtai City from June 2018 to June 2020 as the research object.Among them,92 patients with malignant pleural effusion were regarded as the malignant group,and the remaining 76 patients with benign pleural effusion were regarded as the benign group.The total RNA and total protein of cells in the pleural effusion of the two groups of patients were extracted,and the expressions of LKB1,AMPK and mTOR were detected and compared by immunohistochemistry,and AMPKα,p-AMPKα,LKB1,p-mTOR were detected and compared by Western blotting.LKB1,AMPKαand mTOR mRNA expression were detected and compared using real-time fluorescent quantitative PCR.And the diagnostic value of LKB1,AMPK,mTOR in pleural effusion was analyzed.Results Immunohistochemistry showed that the positive staining of LKB1,AMPK,and mTOR in the pleural effusion of the two groups was brown-yellow,mainly expressed in the cytoplasm.The positive expression rate of LKB1 and AMPK in the malignant group was lower than that of the benign group,and the positive expression rate of mTOR was higher than that of the benign group,and the differences were statistically significant(P<0.05).Western blotting experiments showed that the relative expression of p-AMPKαprotein in the pleural effusion of the two groups had no significant difference(P>0.05).The relative expression of AMPKαand p-mTOR protein in the malignant group was higher than that in the benign group,and the relative expression of LKB1 and mTOR protein was lower than that in the benign group,the differences were statistically significant(P<0.05).Real-time fluorescent quantitative PCR showed that the expression of LKB1 and AMPKαmRNA in the malignant group was lower than that
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