ANALYSIS OF ONCOGENS,TUMOR-SUPPRESSOR GENES,GROWTH FACTORS IN HUMAN HEPATOCELLULAR CARCINOMA AND THEIR APPLICATION TO TREATMENT

建立了4例日本男性肝细胞癌的细胞系，分别命名为OCUH-16、OCUH-A1、OCUH-Nuk1和OCUH-MAS1。它们都具有HCC的形态和染色体特征，本研究使用OCUH-16和OCUH-A1。OCUH-16细胞除表达白蛋白外，还表达转化生长因子-α(TGF-α)。DNA聚合酶-α、c-jun、c-Fos和视网膜母细胞瘤基因产物(PRB)在OCUH-16细胞中也有表达。电镜下可见与细胞膜相关的转化生长因子-α免疫反应，但酶联免疫吸附试验未检测到转化生长因子-α在OCUH-16细胞条件培养液中的表达。逆转录和聚合酶链式反应分析表明，细胞内存在转化生长因子-α信使RNA。在转化生长因子-α中和抗体存在的情况下培养OCUH-16细胞可抑制细胞增殖，并诱导许多细胞发生凋亡。实际上，在人肝细胞癌组织中也检测到了pRb。PRb阳性颗粒见于肝细胞癌的细胞核和肝窦细胞(Kupffer细胞、内皮细胞、淋巴细胞和星形细胞)的胞核中。pRb在早期和晚期肝癌中表达良好，pRb的表达与DNA聚合酶-α的表达有统计学意义的相关性。通过失活转化生长因子-α诱导肝癌细胞凋亡可能适用于临床治疗。

Some cell lines derived from four Japanese men with hepathocellular carcinomas (HCC) were established and designated OCUH-16, OCUH-A1, OCUH-Nuk1, and OCUH-Mas1. All of them have the morphological and chromosomal features of HCC.OCUH-16 and OCUH-A1 were used in this study. OCUH-16 cells were shown to express transforming growth factor-alpha (TGF-alpha) in addition to albumin. DNA polymerase-alpha, c-JUN,c-Fos, and retinoblastoma gene product (pRB) were also expressed in OCUH-16 cells. Electron microscopy revealed TGF-alpha immunoreactivity associated with the cell membrane, but TGF-alpha was not detected in medium conditioned by OCUH-16 cells by enzyme-linked immunosorbent assay. Reverse transcription and polymerase chain reaction analysis revealed the presence of TGF-alpha messenger RNA in the cells. Culture of OCUH-16 cells in the presence of a neutralizing antibody to TGF-alpha inhibited cell proliferation and induced many cells to undergo apoptosis. pRB was also actually detected immunohistochemically in human HCC tissues. The granules positive for pRB were found in the HCC cell nuclei and nuclei of sinusoidal cells (Kupffer cells, endothelial cells, lymphocytes, and stellate cells).pRB expression was well maintained in early and advanced HCC.The expression of pRB statistically correlated with that of DNA polymerase-alpha. The induction of apoptosis in HCC cells by inactivation of TGF-alpha may be applicable to clinical treatment.