Differential sensitivity to the induction of apoptosis by cisplatin in proliferating and quiescent immature rat thymocytes is independent of the levels of drug accumulation and DNA adduct formation.
Immature rat thymocytes readily undergo apoptosis following exposure to many different stimuli, including agents which cause DNA damage, such as the topoisomerase II inhibitor etoposide and irradiation. We have shown previously that cells isolated from the immature rat thymus are resistant to the induction of apoptosis by the DNA-damaging agent cis-diamminedichloroplatinum(II) (cisplatin) (D. L. Evans and C. Dive, Cancer Res., 53:2133-2139, 1993). More than 85% of these thymocytes are quiescent. Here, we demonstrate that following purification of the minority subpopulation of thymocytes that are proliferating, a 2-h exposure to 50 microM cisplatin resulted in rapid apoptosis with 66% apoptotic cells by 12 h. In contrast, purified, nonproliferating thymocytes treated with cisplatin exhibited control levels of apoptosis at 12 h. Both proliferating and nonproliferating thymocytes rapidly underwent apoptosis following continuous exposure to methylprednisolone (10 microM) and etoposide (10 microM). The discrepancy in the levels of apoptosis seen in proliferating and quiescent thymocytes in response to cisplatin could not be attributed to changes in total cellular levels of cisplatin or to the number of DNA-platinum adducts which were determined, respectively, by atomic absorption spectrometry and competitive enzyme-linked immunoadsorbent assay. These results imply that in contrast to engagement of thymocyte apoptosis by methylprednisolone and etoposide, where apoptosis was proliferation independent, cisplatin-induced apoptosis depends on the presence of cells in S and G2-M phases of the cell cycle. Moreover, comparison of etoposide and cisplatin responses in thymocytes suggests that DNA damage per se may not be sufficient to induce apoptosis and that the type of DNA damage is important in this regard.
Evans
D L
DL
Cancer Research Campaign Cellular and Molecular Pharmacology Group, School of Biological Sciences, Manchester University, United Kingdom.
Tilby
M
M
Dive
C
C
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D002458:Cell Fractionation; D002945:Cisplatin / Q000378:metabolism* / Q000493:pharmacokinetics* / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism*; D018736:DNA Adducts; D004249:DNA Damage; D004797:Enzyme-Linked Immunosorbent Assay; D005047:Etoposide / Q000494:pharmacology; D005260:Female; D016195:G2 Phase / Q000187:drug effects / Q000502:physiology; D008297:Male; D008775:Methylprednisolone / Q000494:pharmacology; D008938:Mitosis / Q000187:drug effects / Q000502:physiology; D010984:Platinum / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D016196:S Phase / Q000187:drug effects / Q000502:physiology; D013950:Thymus Gland / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism
D018736:DNA Adducts; C057270:cisplatin-DNA adduct; D010984:Platinum; D005047:Etoposide; D004247:DNA; D002945:Cisplatin; D008775:Methylprednisolone