血清蛋白glypican-3、survivin在肝细胞癌中的表达水平及其临床意义

发布时间：2019-03-01 17:54

【摘要】：肝细胞癌(hepatocellular carcinoma,HCC)是全世界范围内常见的恶性肿瘤之一,其发病率在恶性肿瘤中高居第六位,肿瘤相关死亡率居第三位。肝细胞癌患者生存率低,复发转移率高,而大多数患者在就诊时已属中晚期,其全部5年生存率仅为3-5%且仅有30-40%的HCC患者在确诊时适宜手术,因此早期筛查和诊断对于提高肝细胞癌患者的生存率和临床预后具有重要的意义。人生存素survivin是凋亡抑制蛋白家族中分子最小的成员,近年来的研究发现其在肝癌、胃癌、胰腺癌、前列腺癌等多种肿瘤组织中呈高水平表达,而正常组织中几乎不表达,并且与化疗抵抗、肿瘤复发和肿瘤患者生存期有关。而关于survivin在肝细胞癌患者血清中的表达情况仅有2篇文献,且结论并不明确。磷脂酰肌醇蛋白聚糖-3(Glypican3,GPC3)是硫酸乙酰肝素糖蛋白中磷脂酰肌醇聚糖家族中的一员(GPC1-6),通过C端与糖基磷脂酰基醇的结合而锚定于细胞外膜上,其第358位的精氨酸与359位的丝氨酸发生解离,N端进入血液中,称为可溶性GPC3蛋白。大量研究发现GPC3在调节细胞增殖、分化中起到重要作用,且在胎肝和肝癌组织中特异性高表达,在正常成人组织中几乎无表达。对于GPC3在肝细胞癌血清中表达情况已有多篇文献报道,然而其结论并无一致性。因此,本文将深入研究survivin与GPC3在肝细胞癌患者血清中的表达情况,探讨其对肝细胞癌的诊断价值,并分析比较与肝癌病理分级、TNM分期等临床病理特征的关系,探讨其与预后的关系。目的:1.采用两种ELISA试剂盒对肝细胞癌血清中survivin表达水平进行检测分析,探讨检测血清survivin诊断肝细胞癌的可行性以及筛查一种有价值的诊断试剂盒。2.检测大样本肝细胞癌患者血清中GPC3表达水平,分析比较其对肝细胞癌的诊断精确性,并与常规肝癌肿瘤标志物AFP比较诊断价值方法:1.采用RD和abnova两种试剂盒以酶联免疫吸附法(ELISA)定量检测肝细胞癌、肝硬化、慢性乙型肝炎患者及正常成人血清中survivin蛋白浓度,并分析比较两种试剂盒检测结果的相关性与差异性。2.采用Human GPC3 RD Duoset试剂盒检测890例血清样本(包括肝细胞癌283例,肝硬化267例,慢性乙肝162例,正常对照162例和不典型增生16例)中GPC3蛋白含量,绘制ROC曲线,分析其对肝细胞癌的诊断精确性,并分析比较与肝癌病理分级、TNM分期等临床病理特征的关系,探讨其与预后的关系。结果:1.RD试剂盒检测的80份血清样品中,survivin阳性率为8.75%(7/80),肝细胞癌组阳性率为5%(1/20);对同样的血清样品,abnova试剂盒survivin阳性率为22.5%(18/80),其中肝细胞癌组阳性率为25%(5/20)。且两种试剂盒检测结果均显示HCC组与正常对照组survivin表达水平无明显差异;两种ELISA检测试剂盒对同一份样品检测结果的相关系数r 2为0.0064,,P=0.481,abnova试剂盒结果阳性率高于RD(P=0.002),表明RD和abnova两种试剂盒检测结果存在差异。2.血清GPC3在肝细胞癌、肝硬化、慢乙肝、正常对照和不典型增生中的表达水平分别为0 ng/ml(range=0-14.0ng/ml),0 ng/ml(range=0-0ng/ml),0ng/ml(range=0-12.5ng/ml),0 ng/ml(range=0-1.7ng/ml),和0ng/ml(range=0-4.3ng/ml)。血清GPC3在肝细胞癌(P=0.033)和肝硬化患者(P=0.001)明显高于正常对照,但肝细胞癌与肝硬化患者相比,并无明显差异(P=0.097)。ROC曲线显示血清GPC3诊断肝细胞癌敏感度为39.9%,特异度为60.6%,曲线下面积为0.519,最适cut off值为0.002 ng/ml;血清GPC3表达水平与AFP之间并无明显相关性(P0.05),同时在121例AFP阴性的肝细胞癌患者中有44例GPC3表达水平明显升高。结论:1.血清survivin浓度明显低于两种试剂盒检测下限,此两种试剂盒不适用于检测HCC血清survivin表达水平。2.GPC3可以鉴别肝细胞癌与正常对照,但诊断肝细胞癌的敏感性和特异性较低,与AFP联合诊断,可提高诊断敏感度。
[Abstract]:Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in the world. The survival rate of the patients with hepatocellular carcinoma is low, the rate of recurrence is high, and most of the patients are in the middle and late stage at the time of the visit, and the overall 5-year survival rate is only 3-5% and only 30-40% of the patients with the HCC are suitable for operation at the time of diagnosis, Therefore, early screening and diagnosis are of great significance for improving the survival rate and the clinical prognosis of patients with hepatocellular carcinoma. Survivin is a member of the smallest molecule in the apoptosis-inhibiting protein family. In recent years, it has been found to be a high-level expression in a variety of tumor tissues such as liver cancer, gastric cancer, pancreatic cancer, and prostate cancer. The recurrence of the tumor is related to the survival of the tumor. The expression of survivin in serum of patients with hepatocellular carcinoma is only 2 articles, and the conclusion is not clear. Phospholipid phytate-3 (Glycian3, GPC3) is a member of the glycosaminoglycan family (GPC1-6) in the glyoxaparin glycoprotein, anchored to the outer membrane of the cell by the combination of the C-terminal with the glycolipid-base alcohol, and its 358-position arginine is dissociated from the 359-position serine, The N-terminal enters the blood, known as the soluble GPC3 protein. It is found that GPC3 plays an important role in regulating cell proliferation and differentiation, and is highly expressed in fetal liver and liver cancer tissues, and has little expression in normal adult tissues. There are many literature reports on the expression of GPC3 in the serum of hepatocellular carcinoma, yet its conclusions are not consistent. Therefore, the expression of survivin and GPC3 in the serum of patients with hepatocellular carcinoma is studied in this paper. The diagnostic value of survivin and GPC3 in the serum of hepatocellular carcinoma is discussed, and the relationship between the clinical and pathological features such as the pathological grade and TNM stage of the liver cancer is analyzed and the relation with the prognosis is discussed. Objective:1. The expression level of survivin in the serum of hepatocellular carcinoma was analyzed by two ELISA kits. The feasibility of the detection of survivin in the diagnosis of hepatocellular carcinoma and the screening of a valuable diagnostic kit were also discussed. To detect the level of GPC3 expression in serum of patients with large-sample hepatocellular carcinoma, the diagnostic accuracy of HCC was compared and the diagnostic value of AFP was compared with that of conventional HCC. The expression of survivin protein in the serum of hepatocellular carcinoma, liver cirrhosis, chronic hepatitis B and normal adult was measured by ELISA, and the correlation and difference of the two test cases were compared. The content of GPC3 protein in 890 serum samples (including 283 cases of hepatocellular carcinoma,267 cases of liver cirrhosis,162 cases of chronic hepatitis B,162 cases of normal control and 16 cases of atypical hyperplasia) was detected by using the Human GPC3 RD Duboet kit, and the ROC curve was drawn to analyze its diagnostic accuracy for hepatocellular carcinoma. The relationship between the pathological grade, TNM stage and other clinicopathological characteristics of the liver cancer was analyzed and the relation with the prognosis was discussed. Results:1. The positive rate of survivin was 8.75% (7/80), the positive rate of survivin was 5% (1/20), and the positive rate of survivin was 22.5% (18/80) in the same serum samples and 25% (5/20). The results showed that there was no significant difference in the level of survivin expression in the HCC group and the normal control group. The correlation coefficient r 2 of the two ELISA test kits for the same sample was 0.0064, P = 0.481, and the positive rate of the abnova kit was higher than that of RD (P = 0.002). There is a difference in the results of the detection of both the RD and the abnova kits. The expression levels of serum GPC3 in hepatocellular carcinoma, liver cirrhosis, chronic hepatitis B, normal control and atypical hyperplasia were 0 ng/ ml (range = 0-14.0 ng/ ml),0 ng/ ml (range = 0-0 ng/ ml),0 ng/ ml (range = 0-12.5 ng/ ml),0 ng/ ml (range = 0-1.7 ng/ ml), and 0 ng/ ml (range = 0-4.3 ng/ ml). Serum GPC3 was significantly higher in hepatocellular carcinoma (P = 0.033) and in patients with liver cirrhosis (P = 0.001) than in normal control, but there was no significant difference (P = 0.097). The ROC curve showed that the sensitivity of serum GPC3 in the diagnosis of hepatocellular carcinoma was 39.9%, the specificity was 60.6%, and the area under the curve was 0.519. The best cut off value was 0.002 ng/ ml, and there was no significant correlation between the level of serum GPC3 expression and AFP (P0.05). Conclusion:1. The concentration of survivin was significantly lower than that of the two kits. The two kits were not suitable for detecting the level of survivin expression in HCC.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.7

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