Characteristics and aldehyde dehydrogenase activity of four rat hepatoma cell lines produced by diethylnitrosamine-phenobarbital treatment |
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Authors: | Kwang-Huei Lin Mark F Leach Alvin L Winters Ronald Lindahl |
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Institution: | (1) The Biochemistry Program, Department of Biology, The University of Alabama, 35486 University, Alabama;(2) Microbiology, The University of Alabama, 35486 Alabama, University |
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Abstract: | Summary 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 μM/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-methycholanthrene, 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.
This work was submitted by K.-H. L. in partial fulfillment of the requirements for the Doctor of Philosophy degree in The
Graduate School of The University of Alabama. This work was supported by grant CA-21103 from the National Cancer Institute,
Bethesda, MD. |
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Keywords: | aldehyde dehydrogenase hepatoma cell lines hepatocarcinogenesis |
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