Lin K H, Leach M F, Winters A L, Lindahl R
In Vitro Cell Dev Biol. 1986 May;22(5):263-72. doi: 10.1007/BF02621229.
Recent studies in our laboratory have shown that five established rat hepatoma cell lines provide a wide spectrum of tumor-associated aldehyde dehydrogenase (ALDH) activity representative of the range of activities of this enzyme seen in primary rat hepatocellular carcinomas. Four newly established rat hepatoma cell lines, RLT-2M, RLT-3C, RLT-9F, and RLT-5G, were derived from a primary hepatocellular carcinoma. The primary tumor was induced by a single injection of diethylnitrosamine (15 microM/g body weight) to a 1-d-old female S-D rat followed at weaning by chronic phenobarbital treatment. RLT-2M was established from outgrowths of minced tumor pieces. RLT-3C, RLT-9F, and RLT-5G were cloned from RLT-2M by the serial endpoint dilution. All four lines have been maintained in culture for over 100 passages. The ALDH phenotype in both the primary tumor and the four new cell lines was determined by total activity assay, gel electrophoresis, and histochemistry. By total activity assay, the primary tumor did not possess significant tumor-ALDH activity. In contrast, the four new cell lines expressed tumor-ALDH activity. However, they differed in their basal ALDH activities and in ALDH inducibility by 3-methylcholanthrene, benzo(a)pyrene, and phenobarbital. Additionally, significant decreases in tumor-ALDH activity occurred when cells from each line were passaged in vivo. The four lines have been characterized by light and electron microscopic morphology, tumorigenicity, chromosome number, doubling time, and colony formation efficiency in soft agar.
我们实验室最近的研究表明，五种已建立的大鼠肝癌细胞系具有广泛的肿瘤相关醛脱氢酶（ALDH）活性，代表了在原发性大鼠肝细胞癌中所见该酶活性的范围。四种新建立的大鼠肝癌细胞系，RLT-2M、RLT-3C、RLT-9F和RLT-5G，源自原发性肝细胞癌。原发性肿瘤是通过向1日龄雌性S-D大鼠单次注射二乙基亚硝胺（15微摩尔/克体重）诱导产生的，断奶后进行慢性苯巴比妥治疗。RLT-2M是由切碎的肿瘤块生长而成。RLT-3C、RLT-9F和RLT-5G是通过连续终点稀释从RLT-2M克隆而来。所有四个细胞系已在培养中传代超过100次。通过总活性测定、凝胶电泳和组织化学确定原发性肿瘤和四个新细胞系中的ALDH表型。通过总活性测定，原发性肿瘤不具有显著的肿瘤ALDH活性。相比之下，四个新细胞系表达肿瘤ALDH活性。然而，它们在基础ALDH活性以及对3-甲基胆蒽、苯并(a)芘和苯巴比妥的ALDH诱导性方面存在差异。此外，当每个细胞系的细胞在体内传代时，肿瘤ALDH活性显著降低。这四个细胞系已通过光镜和电镜形态学、致瘤性、染色体数目、倍增时间以及软琼脂中的集落形成效率进行了表征。
