Computerized Video Time-Lapse Microscopy Studies of Ionizing Radiation-Induced Rapid-Interphase and Mitosis-Related Apoptosis in Lymphoid Cells

Abstract Endlich, B., Radford, I.R., Forrester, H.B. and Dewey, W.C. Computerized Video Time-Lapse Microscopy Studies of Ionizing Radiation-Induced Rapid-Interphase and Mitosis-Related Apoptosis in Lymphoid Cells. Computerized video time-lapse (CVTL) microscopy of X-irradiated cultures of cells of the murine lymphoma cell lines ST4 and L5178Y-S and the human lymphoid cell line MOLT-4 demonstrated that these cells exhibit a wide disparity in the timing of induction and execution of radiation-induced cell death that included rapid-interphase apoptosis, delayed apoptosis, and postmitotic apoptosis. ST4 cells that received 2.5 or 4 Gy of X radiation underwent rapid-interphase apoptosis within 2 h. Apoptosis commenced with a 10–20-min burst of membrane blebbing followed by swelling for 2–4 h and cell collapse. No apoptotic bodies were formed. After a dose of 1 Gy, approximately 90% of ST4 cells died by rapid-interphase apoptosis, while the remainder completed several rounds of cell division prior to cell death. Postmitotic death of ST4 cells occurred with the same morphological sequence of events as during rapid-interphase apoptosis induced by doses of 1–4 Gy. In contrast, L5178Y-S and MOLT-4 cells that received 4 Gy underwent apoptosis more slowly, with a complex series of events occurring over 30–60 h. Only 3% of L5178Y-S cells and 24% of MOLT-4 cells underwent apoptosis without attempting cell division. The cells became abnormally large during a long G2-phase delay, and then most of the cells (76–97%) attempted to divide for the first or second time at ∼18–30 h postirradiation. However, either mitosis failed or division was aberrant; i.e., the large cells divided into three or four fragments which eventually fused together. This process was followed by several rounds of complex and unpredictable membrane blebbing, gross distortions of shape, fragmentation–refusion events, and formation of apoptotic bodies, after which the cells collapsed at 36–60 h postirradiation.

[1]  P. Alexander.  Mouse Lymphoma Cells with Different Radiosensitivities , 1961, Nature.

[2]  L. Tolmach,et al.  Time lapse cinemicrographic studies of x-irradiated HeLa S3 cells. I. Cell progression and cell disintegration. , 1969, Biophysical journal.

[3]  L. Thompson,et al.  Proliferation kinetics of x-irradiated mouse L cells studied WITH TIME-lapse photography. II. , 1969, International journal of radiation biology and related studies in physics, chemistry, and medicine.

[4]  A. Wyllie,et al.  Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics , 1972, British Journal of Cancer.

[5]  D. Kemp,et al.  Expression of the immunoglobulin C mu gene in mouse T and B lymphoid and myeloid cell lines. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Cohen,et al.  Gene induction by gamma-irradiation leads to DNA fragmentation in lymphocytes. , 1987, Journal of immunology.

[7]  H. Ohyama,et al.  Radiation-induced interphase death of rat thymocytes is internally programmed (apoptosis). , 1988, International journal of radiation biology and related studies in physics, chemistry, and medicine.

[8]  W. Bodmer,et al.  p53 mutations in colorectal cancer. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[9]  M. Haas,et al.  Frequent mutations in the p53 tumor suppressor gene in human leukemia T-cell lines , 1990, Molecular and cellular biology.

[10]  B. Vogelstein,et al.  Participation of p53 protein in the cellular response to DNA damage. , 1991, Cancer research.

[11]  T. McMillan,et al.  Cellular recovery in two sub-lines of the L5178Y murine leukaemic lymphoblast cell line differing in their sensitivity to ionizing radiation. , 1992, International journal of radiation biology.

[12]  C. Purdie,et al.  Thymocyte apoptosis induced by p53-dependent and independent pathways , 1993, Nature.

[13]  K. Shinohara,et al.  Interphase death and reproductive death in X-irradiated MOLT-4 cells. , 1993, Radiation research.

[14]  V. Chow,et al.  Alternative splicing of the p53 tumor suppressor gene in the Molt-4 T-lymphoblastic leukemia cell line. , 1993, Cancer letters.

[15]  S. Sawada,et al.  Radiation-induced apoptosis and necrosis in Molt-4 cells: a study of dose-effect relationships and their modification. , 1993, International journal of radiation biology.

[16]  Scott W. Lowe,et al.  p53 is required for radiation-induced apoptosis in mouse thymocytes , 1993, Nature.

[17]  H. Tauchi,et al.  Analysis of mitotic cell death caused by radiation in mouse leukaemia L5178Y cells: apoptosis is the ultimate form of cell death following mitotic failure. , 1994, International journal of radiation biology.

[18]  K. Shinohara,et al.  X-ray-induced cell death: apoptosis and necrosis. , 1994, Radiation research.

[19]  I. Radford.  p53 status, DNA double-strand break repair proficiency, and radiation response of mouse lymphoid and myeloid cell lines. , 1994, International journal of radiation biology.

[20]  I. Radford.  Radiation response of mouse lymphoid and myeloid cell lines. Part I. Sensitivity to killing by ionizing radiation, rate of loss of viability, and cell type of origin. , 1994, International journal of radiation biology.

[21]  I. Radford,et al.  Radiation response of mouse lymphoid and myeloid cell lines. Part III. Different signals can lead to apoptosis and may influence sensitivity to killing by DNA double-strand breakage. , 1994, International journal of radiation biology.

[22]  I. Radford,et al.  Radiation response of mouse lymphoid and myeloid cell lines. Part II. Apoptotic death is shown by all lines examined. , 1994, International journal of radiation biology.

[23]  Bert Vogelstein,et al.  Uncoupling of S phase and mitosis induced by anticancer agents in cells lacking p21 , 1996, Nature.

[24]  C. Ling,et al.  Apoptosis induced by X-irradiation of rec-myc cells is postmitotic and not predicted by the time after irradiation or behavior of sister cells. , 1996, Cancer research.

[25]  R. Storer,et al.  The mouse lymphoma L5178Y Tk+/- cell line is heterozygous for a codon 170 mutation in the p53 tumor suppressor gene. , 1997, Mutation research.

[26]  A. Levine.  p53, the Cellular Gatekeeper for Growth and Division , 1997, Cell.

[27]  J. Little,et al.  p53 is involved in but not required for ionizing radiation-induced caspase-3 activation and apoptosis in human lymphoblast cell lines. , 1998, Cancer research.

[28]  K. Kinzler,et al.  Requirement for p53 and p21 to sustain G2 arrest after DNA damage. , 1998, Science.

[29]  I. Radford,et al.  Importance of DNA damage in the induction of apoptosis by ionizing radiation: effect of the scid mutation and DNA ploidy on the radiosensitivity of murine lymphoid cell lines. , 1999, International journal of radiation biology.

[30]  Radford Ir,et al.  Importance of DNA damage in the induction of apoptosis by ionizing radiation: effect of the scid mutation and DNA ploidy on the radiosensitivity of murine lymphoid cell lines. , 1999 .

[31]  C C Ling,et al.  Using computerized video time lapse for quantifying cell death of X-irradiated rat embryo cells transfected with c-myc or c-Ha-ras. , 1999, Cancer research.

[32]  B. Endlich,et al.  Ionizing radiation-induced, Bax-mediated cell death is dependent on activation of cysteine and serine proteases. , 1999, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[33]  K. Takahashi,et al.  Attenuation of caspase-3-dependent apoptosis by Trolox post-treatment of X-irradiated MOLT-4 cells. , 1999, International journal of radiation biology.

[34]  M. Faretta,et al.  Cell cycle perturbations and apoptosis induced by isohomohalichondrin B (IHB), a natural marine compound , 1999, British Journal of Cancer.

[35]  Apoptosis and cancer chemotherapy. , 2001, Trends in cell biology.