Tandem screening of toxic compounds on GFP-labeled bacteria and cancer cells in microtiter plates.
A 96-well fluorescence-based assay has been developed for the rapid screening of potential cytotoxic and bacteriocidal compounds. The assay is based on detection of green fluorescent protein (GFP) in HeLa human carcinoma cells as well as gram negative (Escherichia coli) and gram positive bacteria (Mycobacterium avium). Addition of a toxic compound to the GFP marked cells resulted in the loss of the GFP fluorescence which was readily detected by fluorometry. Thirty-nine distinct naphthoquinone derivatives were screened and several of these compounds were found to be toxic to all cell types. Apart from differences in overall toxicity, two general types of toxic compounds were detected, those that exhibited toxicity to two or all three of the cell types and those that were primarily toxic to the HeLa cells. Our results demonstrate that the parallel screening of both eukaryotic and prokaryotic cells is not only feasible and reproducible but also cost effective.
Montoya
Jessica
J
Department of Biological Sciences, University of Texas at El Paso, El Paso, TX 79968-0519, USA.
Varela-Ramirez
Armando
A
Shanmugasundram
Muthian
M
Martinez
Luis E
LE
Primm
Todd P
TP
Aguilera
Renato J
RJ
D000970:Antineoplastic Agents / Q000494:pharmacology; D001419:Bacteria / Q000166:cytology*; D001681:Biological Assay / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D005470:Fluorometry / Q000379:methods; D015964:Gene Expression Regulation, Bacterial; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D009162:Mycobacterium avium / Q000378:metabolism; D009285:Naphthoquinones / Q000494:pharmacology; D009369:Neoplasms / Q000473:pathology*
D000970:Antineoplastic Agents; D007202:Indicators and Reagents; D009285:Naphthoquinones; D049452:Green Fluorescent Proteins