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

从4名日本男性肝细胞癌（HCC）患者身上提取的细胞系被建立，并命名为OCUH-16、OCUH-A1、OCUH-Nuk1和OCUH-Mas1。它们都具有HCC的形态学和染色体特征。本研究使用OCUH-16和OCUH-A1。OCUH-16细胞除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）也在OCUH-16细胞中表达。电镜显示tgf - α免疫反应性与细胞膜相关，但在OCUH-16细胞条件下的培养基中，酶联免疫吸附试验未检测到tgf - α。逆转录和聚合酶链反应分析显示细胞中存在tgf - α信使RNA。在tgf - α中和抗体的存在下培养OCUH-16细胞抑制细胞增殖并诱导许多细胞凋亡。pRB在人类HCC组织中也被免疫组织化学检测到。pRB阳性颗粒见于HCC细胞核和窦状细胞（Kupffer细胞、内皮细胞、淋巴细胞和星状细胞）细胞核。pRB在早期和晚期HCC中表达良好。pRB的表达与DNA聚合酶α的表达有统计学意义。通过tgf - α失活诱导HCC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.