联合检测尿液微小核糖核酸375和微小核糖核酸145-5p水平在膀胱癌诊断和预后的临床价值

Clinical value of combined detection of urinary miRNA-375 and miR-145-5p levels in the diagnosis and prognosis of bladder cancer

目的观察联合检测尿液微小核糖核酸375(miRNA-375)和微小核糖核酸145-5p(miR-145-5p)水平在膀胱癌诊断和预后的临床价值。方法选择2012年1月至2016年1月在复旦大学附属中山医院青浦分院就诊确诊为膀胱癌的病人112例,为膀胱癌组,选择同期在复旦大学附属中山医院青浦分院行健康体检者45例,为健康对照组。选取两组的尿液标本采用实时荧光定量聚合酶链式反应(qRT-PCR)的方法检测miRNA-375和miR-145-5p的表达水平。观察两组尿液miRNA-375和miR-145-5p水平的差异,膀胱癌病人尿液miRNA-375和miR-145-5p表达水平术前术后变化,与临床指标和预后的关系,及其在诊断膀胱癌和判断其预后的灵敏度和特异度。结果膀胱癌组尿液miRNA-375表达明显高于健康对照组[(3.95±1.26)比(2.30±0.54)P<0.01],术后较术前明显降低[(2.43±1.08)比(3.95±1.26),P<0.01],而miR-145-5p的表达明显低于健康对照组[(1.48±0.75)比(2.68±0.61),P<0.01],术后较术前明显升高[(2.56±0.67)比(1.48±0.75),P<0.01]。尿液miRNA-375和miR-145-5p表达在诊断膀胱癌具有较高的灵敏度和特异度,联合检测的灵敏度87.5%,特异度93.3%,其曲线下面积明显高于miRNA-375(z=2.316,P=0.021)和miR-145-5p(z=2.727,P=0.006)单独检测,而miRNA-375和miR-145-5p之间无明显差别(z=0.921,P>0.05)尿液miRNA-375和miR-145-5p表达水平与性别、年龄、肿瘤部位和肿瘤分级无明显相关性(P>0.05),而与肿瘤分期、淋巴结转移和肿瘤数目具有明显相关性(P<0.01)。膀胱癌病人随访3年,出现23例死亡,89例存活,死亡组的miRNA-375表达水平明显高于存活组(P<0.01),而miR-145-5p表达水平明显低于存活组(P<0.01)。尿液miRNA-375表达水平在预测膀胱癌3年内出现死亡的灵敏度为78.3%,特异度95.5%,其曲线下面积为0.891;而尿液miR-145-5p表达水平在预测膀胱癌3年内出现死亡的灵敏度为95.7%,特异度为70.8%,曲线下面积为0.856。两者联合检测并不能提高预测3年内出现死亡的诊断效能。结论miRNA-375和miR-145-5p参与了膀胱癌的发生发展,尿液中的miRNA-375和miR-145-5p检测对于膀胱癌的诊断和预后判断具有重要临床价值。

Objective To observe the clinical value of combined detection of urinary miRNA-375 and miR-145-5p levels in the diagnosis and prognosis of bladder cancer.Methods 112 patients with bladder cancer in Qingpu Branch of Zhongshan Hospital,Fudan University,from January 2012 to January 2016,were enrolled as the bladder cancer group,and 45people were selected as the healthy control group at the same period.The urine samples in each group were selected to detect the expression levels of miRNA-375 and miR-145-5p by qRT-PCR.The levels of urine miRNA-375 and miR-145-5p levels were observed between bladder cancer group and healthy control group.The urine miRNA-375 and miR-145-5p expression levels in patients with bladder cancer were also observed between before and after operation,and were related with clinical indicators and prognosis.The sensitivity and specificity were also observed in diagnosing bladder cancer and judging its prognosis.Results The expression of urinary miRNA-375 in bladder cancer group was significantly higher than that in healthy control group[(3.95±1.26)vs.(2.30±0.54)P<0.01],and was significantly lower than that before operation[(2.43±1.08)vs.(3.95±1.26),P<0.01],while the expression of miR-145-5p was significantly lower than that in healthy control group[(1.48±0.75)vs.(2.68±0.61),P<0.01],increased significantly compared with before operation[(2.56±0.67)vs.(1.48±0.75),P<0.01].The expression of urinary miRNA-375 and miR-145-5p has high sensitivity and specificity in the diagnosis of bladder cancer,the sensitivity of combined detection was 87.5%and the specificity was 93.3%,the area under the curve was significantly higher than that of miRNA-375(z=2.316,P=0.021)and miR-145-5p(z=2.727,P=0.006),separately,while there was no significant difference between miRNA-375 and miR-145-5p(z=0.921,P>0.05).The expression of miRNA-375 and miR-145-5p was not correlated with gender,age,tumor location and tumor grade(P>0.05),but was significantly correlated with tumor stage,lymph node metastasis and tumor number(P<0.01).Bladder cancer patients were followed up for 3 years,23 deaths occurred,89 patients survived.The expression of miRNA-375 in the death group was significantly higher than that in the survival group(P<0.01),while the expression of miR-145-5p was significantly lower than that in the survival group(P<0.01).The urinary miRNA-375 expression was 78.3%,the specificity was 95.5%,and the area under the curve was 0.891.In predicting death in bladder cancer within 3 years,the sensitivity of urine miR-145-5p expression was 95.7%,specificity was 70.8%,and the area under the curve was 0.856,but the combined detection not improve the diagnostic efficacy inpredicting death within 3 years.Conclusion MiRNA-375 and miR-145-5p are involved in the development of bladder cancer.The detection of miRNA-375 and miR-145-5p in urine has important clinical value for the diagnosis and prognosis of bladder cancer.