Host cell reactivation of cisplatin-damaged plasmid DNA in human non-T leukocyte cell lines.
Host-cell reactivation of cisplatin-damaged pRSVcat was assessed in three B cell lines (SKW 6.4, WIL2-NS, RPMI 1788), the monocytic cell line THP-1, and promyelocytic HL-60 cells. IC50 values following a 3-day exposure of the five leukocyte cell lines to cisplatin ranged from 0.45 to 1.92 microM. Transfer of pRSVcat into all cell lines was effected by electroporation and the resultant CAT activity was measured 24 h later by a rapid single vial CAT assay. CAT activity corresponding to an average of 0.06 units of purified CAT enzyme was expressed by WIL2-NS cells. Very low to no expression of the CAT vector was observed in all other cell lines studied, despite the presence of intracellular levels of 3H-labelled pRSVcat comparable to WIL2-NS. Epstein-Barr virus transformed B cells (SKW 6.4 and RPMI 1788) did not successfully perform host cell reactivation. In WIL2-NS cells, platination of pRSVcat to defined levels of 5-40 platinum molecules per plasmid led to a graded reduction in CAT activity expressed following transfection. Platination levels of 20 and 40 platinum molecules per plasmid did not alter the efficiency of transfer of pRSVcat into these cells by electroporation. Data obtained in this study suggests that EBV transformation may possibly be a negative influence on host cell-reactivation assays for cisplatin-DNA damaged plasmid in non-T human leukocytes.
Dabholkar
M
M
Medicine Branch, National Cancer Institute, Bethesda, Maryland 20892.
Reed
E
E
D001402:B-Lymphocytes; D002460:Cell Line; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D002945:Cisplatin / Q000494:pharmacology*; D004249:DNA Damage; D004260:DNA Repair; D004926:Escherichia coli / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D007962:Leukocytes; D010957:Plasmids; D015183:Restriction Mapping; D014162:Transfection
D015500:Chloramphenicol O-Acetyltransferase; D002945:Cisplatin