Journal Article

Stable expression of rat sulfotransferase 1B1 in V79 cells: activation of benzylic alcohols to mutagens

Wera Teubner, Walter Meinl and Hansruedi Glatt

in Carcinogenesis

Volume 23, issue 11, pages 1877-1884
Published in print November 2002 | ISSN: 0143-3334
Published online November 2002 | e-ISSN: 1460-2180 | DOI: http://dx.doi.org/10.1093/carcin/23.11.1877
Stable expression of rat sulfotransferase 1B1 in V79 cells: activation of benzylic alcohols to mutagens

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We have constructed Chinese hamster V79-derived cell lines (V79-rSULT1B1-A and -B) that express rat sulfotransferase 1B1 (rSULT1B1). Sulfotransferase activity towards 1-naphthol was 1020 ± 220 pmol/min/mg cytosolic protein in V79-rSULT1B1-A cells and 57 ± 9 pmol/ min/mg in V79-rSULT1B1-B cells. These activities were similar over 100 population doublings and at varying cell densities. Immunostaining indicated a cytoplasmatic localization of rSULT1B1. Expression usually was homogeneous within colonies but showed some variation between colonies. The level of rSULT1B1 protein in V79-rSULT1B1-B cells was similar to that in rat liver but higher than in colon mucosa. The cytotoxicity of the benzylic alcohols 4H-cyclopenta[def]chrysen-4-ol and 6-hydroxymethylbenzo-[a]pyrene was enhanced >100-fold in V79-rSULT1B1-A cells compared with SULT-deficient cells (V79p). Likewise, these compounds showed mutagenic effects (at the hprt locus) in V79-rSULT1B1-A cells starting at a concentration of 0.02 and 0.01 μM, respectively, but were inactive in V79p cells even at a concentration of 1 μM. The cell line with the lower expression level, V79-rSULT1B1-B, showed only marginal toxification of the compounds investigated, indicating an important role of the expression level in the test system. A thoroughly characterized mammalian cell system, including positive controls, is now available for studying rSULT1B1-mediated bioactivation of promutagens and protoxicants.

Keywords: BSA, bovine serum albumin; h1B1, human sulfotransferase 1B1; HMBP, 6-hydroxymethylbenzo[a]pyrene; KCP, phosphate-buffered KCl solution; LC50, lethal concentration 50 (leading to a 50% decrease in cell number compared with control cultures); OH-CPC, 4H-cyclopenta[def]chrysen-4-ol; PAH, polycyclic aromatic hydrocarbon; PAPS, 3′-phosphoadenosine-5′-phosphosulfate; PBS, phosphate-buffered saline; PCR, polymerase-chain reaction; r1B1, rat sulfotransferase 1B1; SULT, member of the superfamily of soluble sulfotransferases.

Journal Article.  6991 words.  Illustrated.

Subjects: Clinical Cytogenetics and Molecular Genetics

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