Growth in culture and tumorigenicity after transfection with the ras oncogene of liver epithelial cells from carcinogen-treated rats.
Growth in culture and tumorigenicity after transfection with the ras oncogene of liver epithelial cells from carcinogen-treated rats.
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发表时间：
1987-08
期刊：
影响因子：
11.2
通讯作者：
L. Braun;M. Goyette;P. Yaswen;N. Thompson;N. Fausto
中科院分区：
文献类型：
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作者：
L. Braun;M. Goyette;P. Yaswen;N. Thompson;N. Fausto
两个上皮细胞系命名为LE/2和LE/6建立从细胞离心淘洗从致癌剂处理的大鼠的肝脏。两种细胞系均表现出胎肝细胞的一些特征，如2.3-脱氢酶甲胎蛋白mRNA、醛缩酶A和乳酸脱氢酶4和5的表达。原代培养物含有在体外不增殖的γ-谷氨酰转移酶阳性细胞。第一次传代后，LE/2和LE/6细胞系均为γ-谷氨酰转移酶阴性。通过形态学、非贴壁依赖性生长或裸鼠肿瘤形成测定，两种细胞系均未转化。到第50代，LE/6细胞在存在表皮生长因子的软琼脂中形成大量集落，而在缺乏该生长因子的培养基中没有集落生长。来自5个表皮生长因子诱导的软琼脂集落的克隆细胞群在裸鼠中没有致瘤性。这表明，尽管表皮生长因子反应性晚期传代细胞获得了通常与肿瘤细胞相关的一些表型特性，但这些细胞并未完全转化。LE/6细胞的转化通过转染rasH癌基因（EJ）完成。皮下接种rasH（EJ）转染的LE/6细胞在注射部位产生肿瘤，其组织学特征为中度至高度分化的小梁型肝细胞癌。来自裸鼠肿瘤的肿瘤细胞系为γ-谷氨酰转移酶阳性，并表达甲胎蛋白mRNA。一个克隆细胞系同时表达甲胎蛋白和白蛋白mRNA。这些结果表明，非实质性肝上皮细胞转染一个激活的癌基因可以引起分化的肝细胞肿瘤相似的大鼠肝脏中诱发的致癌性饮食。
Two epithelial cell lines designated LE/2 and LE/6 were established from cells isolated by centrifugal elutriation from the livers of carcinogen-treated rats. Both cell lines exhibit some characteristics of fetal liver cells, such as the expression of the 2.3-kilobase alpha-fetoprotein mRNA, aldolase A, and lactate dehydrogenases 4 and 5. Primary cultures contain gamma-glutamyl transferase-positive cells which do not proliferate in vitro. After the first passage, the LE/2 and LE/6 cell lines are uniformly gamma-glutamyl transferase negative. Neither cell line is transformed as assayed by morphology, anchorage-independent growth, or tumor formation in nude mice. By the 50th passage, LE/6 cells form numerous colonies in soft agar in the presence of epidermal growth factor, while no colonies grow in medium lacking this growth factor. Clonal cell populations derived from five epidermal growth factor-induced soft agar colonies were not tumorigenic in nude mice. This indicates that, although epidermal growth factor-responsive late passage cells had acquired some of the phenotypic properties commonly associated with tumor cells, these cells were not fully transformed. Transformation of LE/6 cells was accomplished by transfection of the rasH oncogene (EJ). Subcutaneous inoculation of rasH (EJ)-transfected LE/6 cells produced tumors at the site of injection with histological features of moderate to well-differentiated trabecular hepatocellular carcinomas. Tumor cell lines derived from the nude mouse tumors are gamma-glutamyl transferase positive and express alpha-fetoprotein mRNA. One clonal cell line expresses both alpha-fetoprotein and albumin mRNA. These results show that nonparenchymal liver epithelial cells transfected with an activated oncogene can give rise to differentiated hepatocellular tumors similar to those induced in livers of rats fed a carcinogenic diet.