Transformation of immortalized woodchuck hepatic cell lines with the c-Ha-ras proto-oncogene.
Transformation of immortalized woodchuck hepatic cell lines with the c-Ha-ras proto-oncogene.
复制标题
用 c-Ha-ras 原癌基因转化永生土拨鼠肝细胞系。
DOI：
10.1093/carcin/17.4.631
复制
发表时间：
1996
期刊：
影响因子：
4.7
通讯作者：
Tennant,BC
中科院分区：
文献类型：
--
作者：
Jacob,JR;Tennant,BC
土拨鼠肝细胞用猴病毒40 T抗原（SV 40）永生化 T-ag）癌基因并用于致癌转化测定。转染 这些细胞系具有激活的c-Ha-ras癌基因（EJ6.6） 导致细胞转化为特征为 软琼脂中的锚定非依赖性生长。转化细胞的集落是 亚克隆和高达80%的癌蛋白表达检测阳性 通过免疫印迹和北方印迹程序。与亲代细胞相比， 行，ras转化的衍生物被改变， 形态学和生长速度。的致瘤潜力 c-Ha-ras转化细胞在重度 联合免疫缺陷（SCID）小鼠。潜伏期1 ~ 4周 在肿瘤被检测到之前， 肿瘤直径达到1 cm。从组织学上看， 经SV 40 T-ag完全转化的株系具有良好的外观， 分化型肝细胞癌（HCC），而来自 c-Ha-ras转化细胞系具有以下外观： 低分化肝癌诱发致癌转化的能力 永生土拨鼠肝细胞系中的事件应提供 有机会研究嗜肝DNA病毒基因的协同作用， 肝癌发生
Woodchuck hepatocytes were immortalized with the simian virus 40 T antigen (SV40 T-ag) oncogene and utilized in an oncogenic transformation assay. Transfection of these cell lines with an activated c-Ha-rasoncogene (EJ6.6) resulted in the transformation of cells to a phenotype characterized by anchorage-independent growth in soft agar. Colonies of transformed cells were subcloned and up to 80% were positive for oncoprotein expression detected by immunoblotand Northern blot procedures. When compared with the parental cell lines,ras-transformed derivatives were altered both morphologically and in growth rate. The tumorigenic potential of c-Ha-rastransformed cells was demonstrated in severe combined immunodeficient (SCID) mice. There was a latency period of 1 to 4 weeks before tumors were detectable and a period of over 7 weeks was required for tumors to reach a diameter of 1 cm. Histologically, tumorsderived from cell lines fully transformed by the SV40 T-ag had the appearance of well differentiated hepatocellular carcinoma (HCC) while tumors derived from c-Ha-rastransformed cell lines had the appearance of poorly differentiated HCC. The capacity to induce oncogenic transformation events in immortalized woodchuck hepatic cell lines should provide the opportunity to study the cooperative effects of hepadnaviral genes in hepatocarcinogenesisin vitro.