Cytotoxicity, anchorage-independent growth, and DNA adduct formation in human neonatal fibroblasts by 1,2,3,4-tetrahydro-7,12-dimethylbenz(a)-anthracene (TH-DMBA), its six aryl fluoro regioisomers, and an exo methylene tautomer.
1,2,3,4-Tetrahydro-7,12-dimethylbenz(a)anthracene (TH-DMBA), its six possible fluorosubstituted regioisomers, and the C-7 exo methylene tautomer of the 11F derivative have been investigated for their cytotoxicity and for their ability to induce anchorage-independent growth and to form adducts in human neonatal foreskin fibroblasts. All compounds tested exhibited a low level of cytotoxicity, determined as percent cloning efficiency, up to a final concentration of 30 micrograms/ml. Except for 5F-TH-DMBA and the C-7 exo methylene tautomer, all compounds induced anchorage-independent growth of neonatal foreskin fibroblasts in soft agar at all concentrations tested (1, 3, 10 and 30 micrograms/ml). The C-7 exo methylene tautomer induced anchorage-independent growth only at a concentration of 10 micrograms/ml. Among the compounds tested the 6F derivative was the most effective compound at 1 microgram/ml. The D-ring fluoro isomers induced anchorage-independent growth at a frequency comparable to TH-DMBA itself, with the 11F derivative being the least effective of the four D-ring regioisomers. All compounds except 5F-TH-DMBA formed detectable adducts with cellular DNA as determined by 32P postlabeling procedures, when the cells were treated at 1 microgram/ml. Two adducts were detected in cells treated with TH-DMBA and four adducts were detected in DNA obtained from cells treated with 6F-TH-DMBA. The level of bonding for the D-ring fluoro isomers was quantitatively less and sometimes qualitatively different than that for TH-DMBA. For the D-ring compounds, the ability to induce anchorage-independent growth frequency correlated with the total quantity of adduct formed. The C-7 exo methylene tautomer formed a single adduct and the level of bonding was less than one adduct per 10(9) nucleotides. Analysis of these results led to the proposal that the planar anthracene ring structure (rings B, C, and D) of TH-DMBA and possibly oxidative metabolism at benzylic carbon 4 of the A-ring are important to DNA bonding and initiation of induction of anchorage-independent growth.
Kumari
H L
HL
Comprehensive Cancer Center, College of Pharmacy, Ohio State University, Columbus.
Milo
G E
GE
Witiak
D T
DT
D015127:9,10-Dimethyl-1,2-benzanthracene / Q000032:analysis / Q000633:toxicity* / Q000031:analogs & derivatives; D015197:Carcinogenicity Tests; D002273:Carcinogens; D002455:Cell Division / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects*; D002478:Cells, Cultured; D004247:DNA / Q000032:analysis*; D018736:DNA Adducts; D005347:Fibroblasts; D006801:Humans; D007231:Infant, Newborn; D007536:Isomerism
C059444:7,12-dimethylbenz(a)anthracene-DNA adduct; D002273:Carcinogens; D018736:DNA Adducts; D015127:9,10-Dimethyl-1,2-benzanthracene; C033113:1,2,3,4-tetrahydro-7,12-dimethylbenz(a)anthracene; D004247:DNA