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Biochem Pharmacol. 2002 Oct 1;64(7):1125-31.
Use of Chlamydomonas reinhardtii mutants for anticancer drug screening.
Maucourt K, Agarwal M, Rene B, Fermandjian S.
Departement de Biologie et Pharmacologie Structurales, LBPA, UMR 8532 CNRS, ENS Cachan, 94235 Cachan, France.
We investigated the possibility of utilizing alga cells instead of mammalian cells for the screening of anticancer drugs. The alga cells grow in synthetic media whereas the mammalian cells require complex and more expensive media along with heavy investment and manpower. To assess the validity of this new approach, analysis of growth inhibition by antitumor agents was carried out jointly on a wall-less (cw15) mutant of Chlamydomonas reinhardtii, that obviates the problem of drug uptake, and the murine leukemic cell line L1210, commonly used for anticancer drug screening. The presence of the topoisomerases I and II (approximately 97 and approximately 2 x 170 kDa, respectively) in the nuclear extracts of C. reinhardtii and their possible role as targets of the drugs was also investigated. Concentrated extracts were separated into >100 and <100 kDa fractions and their topoisomerase I and II activities were measured on relaxation of supercoiled plasmid DNA, decatenation of the catenated kinetoplast DNA and cleavage of plasmid DNA. Our results do not show significant difference in growth inhibition by antitumorals between the wall-less mutant of the alga and the murine leukemic cell line L1210. We noted that alga cells were inhibited by antibiotics that target gyrase, a bacterial variant of topoisomerase II which is also found in chloroplasts. At the molecular level, the alga nuclear fractions, >100 and <100 kDa, displayed the same activities as the mammalian enzymes topoisomerases I and II, respectively, and were blocked by the same poisons. We concluded that the wall-less cw15 mutant of C. reinhardtii could advantageously replace mammalian cells in the screening of the anticancer drugs. The alga enzymes could also provide an opportunity to delineate the phylogeny of the topoisomerase superfamily.
online pharmacy ref. source: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=12234615&dopt=Abstract
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