目的观察原发性肝癌(PHC)患者血清微小RNA(miRNA)-29b的表达,探讨miRNA-29b和甲胎蛋白(AFP)联合检测对PHC的诊断意义。方法收集2007年1月-2010年5月温州医学院附属第-医院外科96名健康志愿者、87例住院PHC患者术前和术后血清,采用定量反转录PCR检测miRNA.29b表达量及ELISA法检测AFP含量,并分析miRNA-29b表达水平与各临床参数间的关系。结果PHC组miRNA.29b[0.250(0.124-0.381)]与健康对照组[0.860(0.587-1.338)]及PHC术后7d组[0.890(0.637~1.414)]比较,表达量明显低(P〈0.001)。PHC组AFP含量[65.4(20.1—212.3)μg/L]与健康对照组[13.3(7.1—19.8)μg/L],及PHC术后7d组[23.2(11.6—55.7)μg/L]比较,表达量明显高(P〈0.001)。miRNA-29b表达水平与病程发展相关,分化程度越低及TNM分期越高其表达水平就越低(P〈0.045和P〈0.001)。Kaplan-Meier结果显示miRNA-29b低表达组的生存率远低于其高表达组,其中位生存期(月)分别为25.52和36.94(P=0.008)。Cox多变量分析显示miRNA.29b水平是影响PHC患者生存率的独立危险因素之一,相对危险度为0.482(95% CI:0.236~0.985)。当ROC曲线确诊PHC的miRNA-29b临界值为0.38和AFP临界值为23.1μg/L时,PHC组血清miRNA-29b、AFP单项检测的敏感度分别为75.9%和70.1%,特异度分别为89.5%和92.7%,两者联合检测的敏感度为87.3%,特异度为88.5%。结论血清miRNA-29b的测定有助于PHC的诊断和预后的估计,联合AFP检测则可有效提高PHC诊断率。
Objective To explore the clinical value of miRNA-29b expression and the combined detection of serum miRNA-29b and alpha-fetoprotein(AFP) in the diagnosis of primary hepatic carcinoma (PHC). Methods From January 2007 to May 2010, the serum levels of miRNA-29b and AFP from 96 healthy controls and 87 PHC patients were measured by quantitative reverse transcription-polymerase chain reaction(qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) respectively. The relationship of miRNA-29b and various clinical parameters was analyzed. Results Serum levels of miRNA-29b in PHC pre-operative group ( 0. 250 ( 0. 124 - 0. 381 ) ) significantly decreased versus the control group [ 0. 860 (0. 587 - 1. 338) ] and the post-operative group (0. 890(0. 637 - 1. 414) ) (P 〈0. 001 ). Also, the levels of AFP in PHC pre-operative group (65.4 (20. 1 -212. 3 ) ) was obviously higher than that in the control group ( 13.3 (7.1 - 19. 8 ) ) and the post-operative group (23.2 ( 11.6 - 55.7 ) ) (P 〈 0. 001 ). A lower expression of miRNA-29b was correlated with lower differentiation and higher TNM stages (P 〈 0. 045, P 〈 0. 001 ). Kaplan-Meier curve analysis revealed that PHC patients with a low serum expression of miRNA-29b had a significantly shortened overall survival when compared with a high serum expression of miRNA-29b(25.52 vs 36. 94 months, P = 0. 008 ). Multivariable Cox regression analysis indicated that the serum expression of miRNA-29b was an independent risk factor for overall survival. Relative risk was 0. 482 (95% confidenceinterval: 0. 236 -0. 985). The critical values for miRNA-29b and AFP were determined at 0. 38 and 23.1 μg/L through the ROC curves. Under the critical value, the sensitivity of miRNA-29b and AFP were 75.9% and 70. 1% and the specificity of miRNA-29b and AFP 89. 5% and 92. 7% respectively. Combined detection could increase the sensitivity up to 87.3%, and achieve a specificity of 88.5% . Conclusion The combined detectio