Lineage-related sensitivity to apoptosis in human tumor cells undergoing hyperthermia.
In this study the role of hyperthermia as an apoptotic trigger was analyzed in four human tumor cell lines: HL60, U937, DOHH2, and K562. These cell lines were chosen because of their well known and different expression of bcl-2 and bcr-abl genes, the expression of which is known to be an antiapoptotic condition. HL60 and U937 cells were strongly susceptible to heat exposure, while DOHH2 cells were weakly sensitive and K562 cells were resistant, thus suggesting a possible gene involvement in this type of programmed cell death. The mechanisms underlying this apoptosis were investigated by flow cytometry, agarose gel electrophoresis, and light and electron microscopy. A subdiploid peak and DNA laddering, both of which are parameters specifically correlated to programmed cell death, were present in HL60 and U937 and, even if less evident, in DOHH2 cells undergoing hyperthermic treatment, and were absent in K562 cells. In addition, DNA single-strand cleavage was revealed by in situ nick translation, observed by confocal microscopy. Morphological analysis confirmed these results and revealed the typical chromatin changes, followed by the appearance of micronuclei and apoptotic bodies.
Falcieri
E
E
Centro di Citometria e Citomorfologia, Università di Urbino, Italy. e.falcieri@uniurb.it
Luchetti
F
F
Burattini
S
S
Canonico
B
B
Santi
S
S
Papa
S
S
D017209:Apoptosis; D002453:Cell Cycle / Q000235:genetics; D019070:Cell Lineage; D004273:DNA, Neoplasm / Q000032:analysis; D004277:DNA, Single-Stranded / Q000378:metabolism; D004587:Electrophoresis, Agar Gel; D005334:Fever / Q000503:physiopathology*; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D020287:In Situ Nick-End Labeling; D020014:K562 Cells; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D014407:Tumor Cells, Cultured / Q000473:pathology* / Q000648:ultrastructure; D020298:U937 Cells
D004273:DNA, Neoplasm; D004277:DNA, Single-Stranded