乙肝病毒相关原发性肝细胞癌的癌前标志物检测

Dectetion of Preneoplastic Markers of Hepatitis B Virus-Associated Hepatocellular Carcinoma

探讨乙肝病毒(HBV)相关原发性肝细胞癌(HCC)癌前标志物的检测及其临床意义。在HBx基因转染的HepG2细胞系中,采用PCR选择性分离cDNA,进行差减杂交,得到5个HBxAg上调基因URG4、URG7、URG11、S15 a、Su i1。建立特异ELISA方法,对中国居民及美国的韩国裔移民共1046例(其中慢性HBV感染400例,对照组646例),检测5个HBxAg上调基因的相应抗体,定其为HBV相关HCC癌前标志物,简称癌前抗体。结果表明,慢性乙肝组、乙肝肝硬化组、HBV相关HCC组以及HCV相关HCC组检出癌前抗体的阳性率与正常人群相比均有显著差异(P<0.01),且差异随慢性乙肝-乙肝肝硬化-肝癌的发展而增大,而HBV携带组、其它肝炎组以及其他肿瘤组与正常人群相比均无显著差异(P>0.05)。在5个癌前抗体中,HBV相关HCC组检出3个或3个以上癌前抗体的阳性率为52.6%(30/57),乙肝肝硬化组为29.2%(35/120),慢性乙肝组为19.0%(29/153),而正常人群仅为0.8%(4/494)。在HBV感染各组病例中,慢性乙肝组癌前抗体阳性率为48.4%(74/153),乙肝肝硬化组为66.7%(80/120),HBV相关HCC组为82.5%(47/57)(P<0.01),平均癌前抗体阳性数分别为1.37、2.12和3.52(P<0.01)。这5个癌前抗体在正常人群中检测不到,而在肝癌及其高危人群中高比例地出现,且阳性率随慢性乙肝-乙肝肝硬化-肝癌的发展而显著升高,反映了这些癌前抗体是肝癌发生的高危风险因子,在肿瘤的发展过程中逐步产生,可以作为癌前病变趋势的诊断依据。

To identify serologic markers that may indicate the early presence of Hepatocellular Carcinoma （HCC） associated with Hepatitis B Virus （HBV）, and analyze their clinical significance. HBV-encoded X （HBx） gene positive and negative HepG2 cells were made and subjected to cDNA subtraction. The 5 host genes, which expression were up-regulated by HBxAg, were named URG4,URG7,URG11,S15a,Sui1. When specific ELISAs were constructed measuring differentially expressed antigens and corresponding antibodies in 1046 individuals from China and American Korean Immigrants（494 HBV-uninfected individuals, 400 HBV-infected individuals and 152 control samples）. Individual antibodies were found in a small percentage of uninfected individuals, in HBV carriers, non- HBV hepatitis patients and other tumor patients. In contrast, antibodies to multiple up-regulated genes were detectable largely in serum samples from patients with chronic HBV hepatitis, HBV-associated cirrhosis and HCC patients（P 〈0.01 ）. The number of antibodies in individual serum samples also differed in these groups. Three or more antibodies were found in 30（52.6% ） of 57 HCC patients, in 35（29.2% ） of 120 patients with cirrhosis, in 29 （19.0%）of 153 patients with HBV hepatitis, and only 4（0.8% ）of 494 uninfected individuals （P 〈 0.01 ）. The positive rate of single antibody were 82.5% , 66.7% and 48.4% in the HCC , cirrhosis and hepatitis patients,and correspondingly the average number of antibodies were 1.37,2.12 和 3.52（P 〈0.01 ）. Hence, antibodies were detected in HBV patients at highest risk for tumor development and in HCC patients. The number of antibodies in a sample was related to increased risk for HCC. This suggests that these antibodies may serve as preneoplastic markers for HCC in HBV-infected patients with chronic liver disease.