The role of apoptosis in cell killing by cisplatin: a flow cytometric study.

Cell killing of L1210 cells by cisplatin has been studied using flow cytometry and DNA gel electrophoresis. Ten hours after a supralethal dose of drug (100 microM), extensive apoptosis was induced. Cells were also susceptible to the induction of apoptosis by nutritional deprivation, for example by incubation in arginine-deficient medium. After treatment in full medium with doses of drug in the range 1-10 microM, cells experienced a slow-down in S-phase transit followed by a G2 block. Cells either overcame the G2 block and continued to cycle or enlarged and eventually died. There was no evidence to suggest that cells dying from the G2 block underwent apoptosis. The data were consistent with a dual mechanism of cell death-higher doses of drug led to rapid death through apoptosis; lower doses led to death at later times resulting from failure to overcome a block in G2.

[1]  A. Eastman,et al.  Activation of programmed cell death (apoptosis) by cisplatin, other anticancer drugs, toxins and hyperthermia. , 1990, Biochemical pharmacology.

[2]  F. Cope,et al.  Apoptosis in C3H/10T1/2 mouse embryonic cells: evidence for internucleosomal DNA modification in the absence of double-strand cleavage. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[3]  L. Thompson,et al.  Defective removal of DNA cross-links in a repair-deficient mutant of Chinese hamster cells. , 1982, Cancer research.

[4]  D L Evans,et al.  Analysis and discrimination of necrosis and apoptosis (programmed cell death) by multiparameter flow cytometry. , 1992, Biochimica et biophysica acta.

[5]  A. Wyllie,et al.  Cell death: the significance of apoptosis. , 1980, International review of cytology.

[6]  Z. Darżynkiewicz,et al.  Changes in nuclear chromatin related to apoptosis or necrosis induced by the DNA topoisomerase II inhibitor fostriecin in MOLT-4 and HL-60 cells are revealed by altered DNA sensitivity to denaturation. , 1992, Experimental cell research.

[7]  W. Telford,et al.  Evaluation of glucocorticoid‐induced DNA fragmentation in mouse thymocytes by flow cytometry , 1991, Cell proliferation.

[8]  J. J. Roberts,et al.  REPAIR OF cis -PLATINUM (II) DIAMMINE DICHLORIDE-INDUCED DNA DAMAGE AND CELL SENSITIVITY , 1980 .

[9]  W. Telford,et al.  Comparative evaluation of several DNA binding dyes in the detection of apoptosis-associated chromatin degradation by flow cytometry. , 1992, Cytometry.

[10]  E. Pettersen,et al.  Synergistic cell inactivation by cis-dichlorodiammineplatinum in combination with 1-propargyl-5-chloropyrimidin-2-one. , 1987, British Journal of Cancer.

[11]  J. Cidlowski,et al.  Analysis of glucocorticoid actions on rat thymocyte deoxyribonucleic acid by fluorescence-activated flow cytometry. , 1988, Endocrinology.

[12]  T. Tsuji,et al.  Flow cytometric analysis of the kinetic effects of cisplatin on lung cancer cells. , 1989, Cytometry.

[13]  J. Roberts,et al.  Effects of cis-platinum(II) diamminedichloride on survival and the rate of DNA synthesis in synchronously growing HeLa cells in the absence and presence of caffeine. , 1978, Chemico-biological interactions.

[14]  M. Ormerod,et al.  Cell cycle analysis of asynchronous cell populations by flow cytometry using bromodeoxyuridine label and Hoechst-propidium iodide stain. , 1992, Cytometry.

[15]  A. Eastman.  The formation, isolation and characterization of DNA adducts produced by anticancer platinum complexes. , 1987, Pharmacology & therapeutics.

[16]  P. Rabinovitch,et al.  BrdU-Hoechst flow cytometry: a unique tool for quantitative cell cycle analysis. , 1988, Experimental cell research.

[17]  I Nicoletti,et al.  A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. , 1991, Journal of immunological methods.

[18]  M. Collins,et al.  Regulation of apoptosis in interleukin‐3‐dependent hemopoietic cells by interleukin‐3 and calcium ionophores. , 1990, The EMBO journal.

[19]  X. M. Sun,et al.  Key morphological features of apoptosis may occur in the absence of internucleosomal DNA fragmentation. , 1992, The Biochemical journal.

[20]  G. Bastian,et al.  BrdUrd/DNA flow cytometry analysis demonstrates cis-diamminedichloroplatinum (II)-induced multiple cell-cycle modifications on human lung carcinoma cells. , 1992, Cytometry.

[21]  Z. Darżynkiewicz,et al.  Features of apoptotic cells measured by flow cytometry. , 1992, Cytometry.

[22]  T. L. Jones,et al.  Quantitative determination of the induction of apoptosis in a murine B cell line using flow cytometric bivariate cell cycle analysis. , 1992, Cellular immunology.

[23]  X. M. Sun,et al.  A flow-cytometric method for the separation and quantitation of normal and apoptotic thymocytes. , 1992, Analytical biochemistry.

[24]  J. Roberts.  The Repair of DNA Modified by Cytotoxic, Mutagenic, and Carcinogenic Chemicals , 1978 .

[25]  C. Sorenson,et al.  Analysis of events associated with cell cycle arrest at G2 phase and cell death induced by cisplatin. , 1990, Journal of the National Cancer Institute.

[26]  J. Hickman,et al.  Drug-target interactions: only the first step in the commitment to a programmed cell death? , 1991, British Journal of Cancer.

[27]  H. Folch,et al.  Protein kinase-C involvement in thymocyte apoptosis induced by hydrocortisone. , 1990, Cellular immunology.

[28]  X. M. Sun,et al.  Increased membrane permeability of apoptotic thymocytes: a flow cytometric study. , 1993, Cytometry.

[29]  A. Eastman.  Interstrand cross-links and sequence specificity in the reaction of cis-dichloro(ethylenediamine)platinum(II) with DNA. , 1985, Biochemistry.

[30]  M. Collins,et al.  Apoptosis in interleukin-3-dependent haemopoietic cells. Quantification by two flow cytometric methods. , 1992, Journal of immunological methods.

[31]  P. Walker,et al.  Topoisomerase II-reactive chemotherapeutic drugs induce apoptosis in thymocytes. , 1991, Cancer research.

[32]  Z. Darżynkiewicz,et al.  Apoptosis of rat thymocytes triggered by prednisolone, camptothecin, or teniposide is selective to G0 cells and is prevented by inhibitors of proteases. , 1992, Oncology research.

[33]  A. Wyllie,et al.  Apoptosis: mechanisms and roles in pathology. , 1991, International review of experimental pathology.