血清miR-143和miR-145水平诊断肝癌的临床价值研究

发布时间：2018-03-24 12:48

本文选题：肝癌　切入点：miR-143　出处：《广西医科大学》2017年硕士论文

【摘要】：目的原发性肝癌(Hepatocellular carcinoma)全球发病率逐年增长,在肿瘤相关死亡中位居第二位。如能对原发性肝癌进行早期的诊断,就能使其治疗方案得到很好的效果,这对于提高患者的生活质量,并且延长其生存期都有很大帮助。现今病理学检查、影像学检查和血清甲胎蛋白(alpha-feto protein,AFP)检测等作为临床上主要的诊断肝癌的方法,虽然AFP检测特异性高,但有30%~40%的肝癌患者呈阴性。所以,各科研人员也认识到不断找寻更好更新的肝癌血清肿瘤标记物非常有必要。到目前为止,国内针对miR-143、miR-145在肝癌表达情况的研究较少。本研究探讨了肝癌患者血清中的miR-143、miR-145表达水平,并阐明血清miR-143、miR-145与肝癌标记物AFP、AST和ALT的关系,探讨miR-143、miR-145与AFP联合检测在肝癌患者中的诊断价值,为HCC临床诊断提供新的有效的分子标志物。方法符合要求的肝癌、肝硬化患者各30例和30例同期健康对照的血清均被纳入本研究,所有纳入人群清晨空腹静脉血离心得到上清液,按总RNA提取试剂盒说明书提取细胞总RNA,按逆转录试剂盒说明书将总RNA逆转录成c DNA,应用实时荧光定量PCR方法检测在肝癌、肝硬化患者和健康体检者血清中的miR-143、miR-145的表达水平,所得实验结果采用相对定量法进行分析,miRNA表达量以2-△△Ct表示。使用受试者工作特征(ROC)曲线和曲线下面积(AUC)分析miR-143、miR-145表达水平和AFP联合检测对肝癌诊断的灵敏度和特异性,并确定最佳诊断阈值,miR-143、miR-145与AFP的一致性检验采用Kappa分析。结果miR-143和miR-145在肝癌患者血清表达水平最高,肝硬化次之,正常人最低,3组间比较差异有统计学意义(P0.05)。不同临床分期肝癌患者其miR-143和miR-145表达水平无统计学差异(P0.05)。肝癌患者血清miR-143、miR-145表达水平与AFP、AST、ALT无显著相关性(P0.05)?miR-143诊断肝癌的AUC为0.86,最佳阈值为1.30,敏感度为0.871,特异度为0.733。miR-145诊断肝癌的AUC为0.70,最佳阈值为1.05,敏感度为0.806,特异度为0.600。miR-143+AFP、miR-145+AFP、miR-143+miR-145+AFP对肝癌诊断效能均较高,其中miR-143+miR-145+AFP的诊断效能最高(AUC=0.994),但miR-143+AFP的诊断敏感度最高(敏感度=0.903)。结论1、miR-143和miR-145在肝癌患者血清表达水平最高。2、miR-143、miR-145诊断肝癌的敏感度与特异度均较高。提示:miR-143、miR-145与肝癌的发病密切相关,并可作为肝癌有效的诊断标记物,联合检测miR-143、miR-145及AFP可以更有效的检测肝癌。
[Abstract]:Objective the global incidence of primary liver cancer (HCC) has been increasing year by year, and it ranks second in tumor-related deaths. It is of great help to improve the quality of life of patients and prolong their survival. Nowadays, pathological examination, imaging examination and serum alpha-feto protein protein (AFP) test are the main methods for the diagnosis of liver cancer. Although the specificity of AFP detection is high, 30% of HCC patients are negative. Therefore, researchers also realize that it is necessary to constantly search for better updated tumor markers in liver cancer serum. The expression of miR-143 miR-145 in serum of patients with liver cancer was studied, and the relationship between miR-143 miR-145 and liver cancer markers AFPNAST and ALT was elucidated. To explore the diagnostic value of miR-143 miR-145 combined with AFP in patients with liver cancer, and to provide a new and effective molecular marker for clinical diagnosis of HCC. Serum samples of 30 patients with liver cirrhosis and 30 healthy controls were included in this study. The supernatants were obtained by early morning fasting venous blood centrifugation in all subjects. Total RNA was extracted according to the instructions of the total RNA extraction kit, and the total RNA was reverse transcribed into c DNA according to the instructions of the reverse transcription kit. The expression of miR-143 miR-145 in serum of patients with liver cancer, liver cirrhosis and healthy persons was detected by real-time fluorescence quantitative PCR method. The results showed that the expression of miRNA was expressed as 2Ct by relative quantitative method. The expression level of miR-143 miR-145 and the sensitivity and specificity of combined detection of miR-143 miR-145 and AFP in the diagnosis of hepatocellular carcinoma were analyzed by using the roc curve and area under the curve. The best diagnostic threshold was determined. The consistency test between miR-143 miR-145 and AFP was analyzed by Kappa. Results the expression of miR-143 and miR-145 in serum of patients with liver cancer was the highest, followed by cirrhosis. There was no significant difference in the expression of miR-143 and miR-145 in patients with different clinical stages of liver cancer. There was no significant correlation between the expression of miR-143 miR-145 in serum and the level of alt in patients with liver cancer. The AUC of miR-143 was 0.86, the best threshold was 1.30, the sensitivity was 0.871, the specificity was 0.70 for 0.733.miR-145, the best threshold was 1.05, the sensitivity was 0.806, and the specificity was 0.600.miR-143 AFPmmiR-145 AFPmiR-143 miR-145 AFP for the diagnosis of HCC. The diagnostic efficacy of miR-143 miR-145 AFP was the highest, but the sensitivity of miR-143 AFP was the highest (sensitivity: 0.903). Conclusion\\\%\\%\%\% miR-143\%\%\%\% miR-@@. Is closely related to the onset of the disease, The combined detection of miR-143 miR-145 and AFP can be used as an effective diagnostic marker for HCC.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7

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