Shiota Goshi, Oyama Kenji, Udagawa Akihide, Nomi Takahiro, Tanaka Kiwamu, Tsutsumi Atsushi, Noguchi Noya, Okano Jun-ichi, Kishimoto Yosuke, Kanbe Takamasa, Kishimoto Hiroyuki, Kawasaki Hironaka
Department of Genetic Medicine and Regenerative Therapeutics, Graduate School of Medicine, Tottori University, Japan.
Hepatogastroenterology. 2009 Sep-Oct;56(94-95):1516-20.
It has been reported that hepatitis B virus (HBV) DNA is detected in serum and/or liver in patients with hepatocellular carcinoma (HCC) without HBsAg. To adress this issue, we analyzed HBV genome in 2 HCC cases without HBsAg. The DNA from serum from patients with HCC was amplified with a nested PCR, and 'a' determinant of S region, core promoter region and precore region were sequenced. The first case, a 50 years-old male, was negative for HBsAg and HBeAg, and positive for anti-HBs, anti-HBe and anti-HBc. Viral load of HBV in serum was 4.0 log genome equivalent/ml by TMA assay, and was 1.1 X 105 copy/ml by real-time PCR system. A nucleotide analysis of the common 'a' determinant of S gene showed that the 5 first amino acids of 'a' determinant, CTIPA, were changed to CKTCTTPA. The second case, a 76 years-old male, was positive for anti-HBe, but negative for HBsAg, anti-HBs, HBeAg and anti-HBc. No missense or nonsense mutations were seen in 'a' determinant of S region. Viral load of serum HBV was < 3.7 log genome equivalent/ml by TMA assay, but was 2.4X103 copy/ml by real-time PCR system. The results of the present study suggest that the mechanisms of HBsAg loss are diverse among HCC patients without HBsAg, and that an analysis of HBV genome is a useful tool to dissolve molecular mechanisms losing HBs antigenicity.
据报道，在无乙肝表面抗原（HBsAg）的肝细胞癌（HCC）患者的血清和/或肝脏中检测到乙肝病毒（HBV）DNA。为解决这一问题，我们分析了2例无HBsAg的HCC病例中的HBV基因组。用巢式聚合酶链反应（PCR）扩增HCC患者血清中的DNA，并对S区的“a”决定簇、核心启动子区和前核心区进行测序。第一例患者为50岁男性，HBsAg和HBeAg均为阴性，抗-HBs、抗-HBe和抗-HBc均为阳性。通过转录介导的扩增（TMA）检测，血清中HBV的病毒载量为4.0 log基因组当量/ml，通过实时PCR系统检测为1.1×105拷贝/ml。对S基因常见“a”决定簇的核苷酸分析显示，“a”决定簇的前5个氨基酸CTIPA变为CKTCTTPA。第二例患者为76岁男性，抗-HBe阳性，但HBsAg、抗-HBs、HBeAg和抗-HBc均为阴性。在S区的“a”决定簇中未发现错义或无义突变。通过TMA检测，血清HBV的病毒载量<3.7 log基因组当量/ml，但通过实时PCR系统检测为2.4×103拷贝/ml。本研究结果表明，在无HBsAg的HCC患者中，HBsAg缺失的机制多种多样，并且对HBV基因组的分析是解析丧失HBs抗原性分子机制的有用工具。
