Kim M Y, Park E, Park J H, Park D H, Moon W S, Cho B H, Shin H S, Kim D G
Department of Internal Medicine, Division of GI and Hepatology, Research Institute of Clinical Medicine, Chonbuk National University Medical School and Hospital, Chonju, Chonbuk 561-712, Republic of Korea.
Oncogene. 2001 Jul 27;20(33):4568-75. doi: 10.1038/sj.onc.1204626.
Chronic hepatitis B virus (HBV) infection is known to be one of the major causes in the development of hepatocellular carcinoma (HCC), although the biomolecular mechanism(s) involved remain unclear. To identify the cellular gene(s) involved in HBV-associated hepatocarcinogenesis, we used the mRNA differential display method and examined three paired tumor and nontumor tissues, all of which had chromosomally integrated HBV-DNA through chronic infection. Using 240 different combinations of three one-base anchored oligo-dT primers and 80 arbitrary 13-mers, genes decreased or increased in expression more than twofold between each tumor tissue and its paired nontumor tissue were identified. Twenty-nine known genes and four novel genes were differentially over-expressed in the HCC tumor tissues. In contrast, 27 known genes and five novel genes were under-expressed in those tumor tissues. The nucleotide sequences of the nine novel gene fragments were determined and their expression patterns were examined in 40 HCC samples. HA61T2, PT18, HG63T1, and HG57T1 were preferentially over-expressed in 32 cases (80%, P<0.001), 24 cases (60%), 23 cases (57.5%) and 22 cases (55%) of the 40 tumor tissues, respectively. There was an increased frequency of HG57T1 over-expression in HCC patients with HBV-positive serology and low serum alpha-feto protein (AFP) levels (P<0.05). DNT10, PT8, PT19, ENT25 and HA6T4 were under-expressed in 26 cases (65%), 23 cases (57.5%), 21 cases (53%), 20 cases (50%) and 18 cases (45%) of the 40 tumor samples, respectively. There was a strong correlation of DNT10 under-expression with high serum AFP level in HCC patients, irrespective of HBV serology (P<0.01). HA6T4 was preferentially under-expressed in HCC tumors in patients with HBV-positive serology and high serum AFP levels (P<0.05). Thus, the functional analyses of the known and novel genes identified in this study should prove valuable to further understand the mechanism(s) of hepatocarcinogenesis.
慢性乙型肝炎病毒（HBV）感染是肝细胞癌（HCC）发生发展的主要原因之一，尽管其中涉及的生物分子机制仍不清楚。为了鉴定与HBV相关肝癌发生有关的细胞基因，我们使用了mRNA差异显示方法，检测了三对肿瘤组织和非肿瘤组织，所有这些组织均通过慢性感染而发生了染色体整合的HBV-DNA。使用三种单碱基锚定的oligo-dT引物和80种任意的13聚体的240种不同组合，鉴定出在每个肿瘤组织与其配对的非肿瘤组织之间表达降低或升高超过两倍的基因。29个已知基因和4个新基因在HCC肿瘤组织中差异过表达。相反，27个已知基因和5个新基因在这些肿瘤组织中表达不足。测定了9个新基因片段的核苷酸序列，并在40个HCC样本中检测了它们的表达模式。HA61T2、PT18、HG63T1和HG57T1在40个肿瘤组织中分别有32例（80%，P<0.001）、24例（60%）、23例（57.5%）和22例（55%）优先过表达。在血清学HBV阳性且血清甲胎蛋白（AFP）水平低的HCC患者中，HG57T1过表达的频率增加（P<0.05）。DNT10、PT8、PT19、ENT25和HA6T4在40个肿瘤样本中分别有26例（65%）、23例（57.5%）、21例（53%）、20例（50%）和18例（45%）表达不足。在HCC患者中，无论HBV血清学情况如何，DNT10表达不足与高血清AFP水平密切相关（P<0.01）。HA6T4在血清学HBV阳性且血清AFP水平高的HCC肿瘤中优先表达不足（P<0.05）。因此，本研究中鉴定出的已知基因和新基因的功能分析对于进一步了解肝癌发生机制应具有重要价值。
