Clonogenicity of human leukemic cells protected from cell-lethal agents by heat shock protein 70.
Pretreatment of human leukemia THP-1 cells with heat shock protein Hsp70 (Hsp70) protected them from the cell-lethal effects of the topoisomerase II inhibitor, lucanthone and from ionizing radiation. Cell viability was scored in clonogenic assays of single cells grown in liquid medium containing 0.5% methyl cellulose. Colonies were observed and rapidly scored after staining with the tetrazolium salt, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide. The frequency of abasic sites in the deoxyribonucleic acid (DNA) of THP-1 cells was reduced when these cells were treated with Hsp70. Hsp70 is presumed to have protected the cells by promoting repair of cell DNA, in agreement with previous studies that showed that Hsp70 enhanced base excision repair by purified enzymes. The shoulders of radiation dose-response curves were enhanced by pretreatment of cells with Hsp70 and, importantly, were reduced when cells were transfected with ribonucleic acid designed to silence Hsp70. Hsp70 influenced repair of sublethal damage after radiation.
Bases
Robert
R
Departments of Radiology and Radiation Oncology, Montefiore Medical Center, Albert Einstein College of Medicine, Bronx, NY 10467, USA. rbases@worldnet.att.net
D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D003114:Colony-Forming Units Assay; D019610:Cytoprotection; D004249:DNA Damage / Q000187:drug effects / Q000528:radiation effects; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005720:Gamma Rays / Q000009:adverse effects*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D007948:Leukemia, Monocytic, Acute / Q000473:pathology; D008154:Lucanthone / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000378:metabolism
D004791:Enzyme Inhibitors; D018840:HSP70 Heat-Shock Proteins; D034741:RNA, Small Interfering; D008154:Lucanthone