Coexpression of PCNA and p21 for DNA repair in small intestinal crypt cells of mouse with 60Co γ-rays irradiation

방사선을 조사한 마우스의 소장 음와세포에서 DNA 수복을 위한 PCNA와 p21의 발현 양상

  • Hong, Suji (Department of Veterinary Medcine, College of Applied Life Sciences and Applied Radiological Science Research Institute, Cheju National University) ;
  • Hwang, Insun (Department of Veterinary Medcine, College of Applied Life Sciences and Applied Radiological Science Research Institute, Cheju National University) ;
  • Ahn, Meejung (Department of Veterinary Medcine, College of Applied Life Sciences and Applied Radiological Science Research Institute, Cheju National University) ;
  • Shin, Taekyun (Department of Veterinary Medcine, College of Applied Life Sciences and Applied Radiological Science Research Institute, Cheju National University) ;
  • Joo, Hong-gu (Department of Veterinary Medcine, College of Applied Life Sciences and Applied Radiological Science Research Institute, Cheju National University) ;
  • Park, HyunJeong (Department of Veterinary Medcine, College of Applied Life Sciences and Applied Radiological Science Research Institute, Cheju National University) ;
  • Jee, Youngheun (Department of Veterinary Medcine, College of Applied Life Sciences and Applied Radiological Science Research Institute, Cheju National University)
  • 홍수지 (제주대학교 농업생명과학대학 수의학과 방사선응용과학연구소) ;
  • 황인선 (제주대학교 농업생명과학대학 수의학과 방사선응용과학연구소) ;
  • 안미정 (제주대학교 농업생명과학대학 수의학과 방사선응용과학연구소) ;
  • 신태균 (제주대학교 농업생명과학대학 수의학과 방사선응용과학연구소) ;
  • 주홍구 (제주대학교 농업생명과학대학 수의학과 방사선응용과학연구소) ;
  • 박현정 (제주대학교 농업생명과학대학 수의학과 방사선응용과학연구소) ;
  • 지영흔 (제주대학교 농업생명과학대학 수의학과 방사선응용과학연구소)
  • Accepted : 2005.11.22
  • Published : 2005.12.29

Abstract

The irradiation of radioactive ${\gamma}-ray$ induces apoptosis of radiosensitive organs for homeostasis. In this study, we investigated the repair mechanisms for homeostasis in the small intestine after cell damage by $^{60}Co\;{\gamma}-ray$ irradiation. The apoptosis was most frequently observed in the crypt cells of the small intestine after four and six hours by radioactive ${\gamma}-ray$ irradiation, and the frequency of apoptosis was proportional to the amount of irradiation. Also, the number of apoptotic cells was coincident with expression pattern of p53. Interestingly, PCNA (proliferating cell nuclear antigen) which is engaged in DNA replication and repair was expressed in apoptotic cells of small intestinal crypts. Also, it was observed that cell-cycle regulator p21 which is known to induce cell-cycle arrest is co-expressed in the same apoptotic cells of irradiated small intestinal crypt cells. These findings suggest that the co-expression of PCNA and p21 proteins, which may lead to resistance to DNA damage through cell-cycle arrest is closely associated with repair of damaged gastrointestinal cells after ${\gamma}-ray$ irradiation.

Keywords

References

  1. Ando T, Kawabe T, Ohara H, Docummun B, Itoh M, Okamoto T. Involvement of the interaction between p21 and proliferating cell nuclear antigen for the maintenance of G2/M arrest after DNA damage. J Biol Chem 2001, 276, 42971-42977 https://doi.org/10.1074/jbc.M106460200
  2. Brugarolas J, Chandrasekaran C, Gordon JI, Beach D, Jacks T, Hannon GJ. Radiation-induced cell cycle arrest compromised by p21 deficiency. Nature 1995, 377, 552-557 https://doi.org/10.1038/377552a0
  3. Clarke AR, Gledhill S, Hooper ML, Bird CC, Wyllie AH. p53 dependence of early apoptotic and proliferation responses within the mouse intestinal epithelium following gamma-irradiation. Oncogene 1994, 9, 1767-1773
  4. Clarke AR, Howard LA, Harrison DJ, Winton DJ. p53 mutation frequency and apoptosis in the murine small intestine. Oncogene 1997, 14, 2015-2018 https://doi.org/10.1038/sj.onc.1201040
  5. Dulic V, Kaufmann WK, Wilson SJ, Tlsty TD, Lees E, Harper JW, Elledge SJ, Reed SI. p53-dependent inhibition of cyclin-dependent kinase activities in human fibroblasts during radiation-induced G1 arrest. Cell 1994, 76, 1013-1023 https://doi.org/10.1016/0092-8674(94)90379-4
  6. El-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B. WAF1, a potential mediator of p53 tumor suppression. Cell 1993, 75, 817-825 https://doi.org/10.1016/0092-8674(93)90500-P
  7. Elledge SJ. Cell cycle checkpoints: preventing an identity crisis. Science 1996, 274, 1664-1672 https://doi.org/10.1126/science.274.5293.1664
  8. Gavrieli Y, Sherman Y, Ben-Sasson SA. Identification of programmed cell death In situ via specific labelling of nuclear DNA fragmentation. J Cell Biol 1992, 119, 493-501 https://doi.org/10.1083/jcb.119.3.493
  9. Goodhead DT. Mechanisms for the biological effectiveness of high-LET radiation. J Radiat Res (Tokyo) 1999, 40, 1-13
  10. Goubin F, Ducommun B. Identification of binding domains on the p21Cip1 cyclin-dependent kinase inhibitor. Oncogene 1995, 10, 2281-2287
  11. Hartwell LH, Weinert TA. Checkpoints: controls that ensure the order of cell cycle events. Science 1989, 246, 629-634 https://doi.org/10.1126/science.2683079
  12. Hindges R, Hubscher U. DNA polymerase delta, an essential enzyme for DNA transactions. Biol Chem 1997, 378, 345-362
  13. Kastan MB, Onyekwere O, Sidransky D, Vogelstein B, Craig RW. Participation of p53 protein in the cellular response to DNA damage. Cancer Res 1991, 51, 6304-6311
  14. Kastan MB, Radin AI, Kuerbitz SJ, Onyekwere O, Wolkow CA, Civin CI, Stone KD, Woo T, Ravindranath Y, Craig RW. Levels of p53 protein increase with maturation in human hematopoietic cells. Cancer Res 1991, 51, 4279-4286
  15. Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implication in the tissue kinetics. Br J cancer 1972, 26, 239-257 https://doi.org/10.1038/bjc.1972.33
  16. Lee JM, Bernstein A. p53 mutations increase resistance to ionizing radiation. Proc Natl Acad Sci USA 1993, 90, 5742-5746
  17. Merritt AJ, Allen TD, Potten CS, Hickman JA. Apoptosis in small intestinal epithelial from p53-null mice: evidence for a delayed, p53-independent G2/Massociated cell death after gamma-irradiation. Oncogene 1997, 14, 2759-2766 https://doi.org/10.1038/sj.onc.1201126
  18. Merritt AJ, Potten CS, Kemp CJ, Hickman JA, Balmain A, Lane DP, Hall PA. The role of p53 in spontaneous and radiation-induced apoptosis in the gastrointestinal tract of normal and p53-deficient mice. Cancer Res 1994, 54, 614-617
  19. Paunesku T, Mittal S, Protic M, Oryhon J, Korolev SV, Joachimiak A, Woloschak GE. Proliferating cell nuclear antigen (PCNA): ringmaster of the genome. Int J Radiat Biol 2001, 77, 1007-1021 https://doi.org/10.1080/09553000110069335
  20. Prelich G, Tan CK, Kostura M, Mathews MB, So AG, Downey KM, Stillman B. Functional identity of proliferating cell nuclear antigen and a DNA polymerasedelta auxiliary protein. Nature 1987, 326, 517-520 https://doi.org/10.1038/326517a0
  21. Prosperi E. Multiple roles of the proliferating cell nuclear antigen: DNA replication, repair and cell cycle control. Prog Cell Cycle Res 1997, 3, 193-210
  22. Selvakumaran M, Lin HK, Miyashita T, Wang HG, Krajewski S, Reed JC, Hoffman B, Liebermann D. Immediate early up-regulation of bax expression by p53 but not TGF beta 1: a paradigm for distinct apoptotic pathways. Oncogene 1994, 9, 1791-1798
  23. Szepesi A, Gelfand EW, Lucas JJ. Association of proliferating cell nuclear antigen with cyclin-dependent kinases and cyclins in normal and transformed human T lymphocytes. Blood 1994, 84, 3413-3421
  24. Tan CK, Castillo C, So AG, Downey KM. An auxiliary protein for DNA polymerase-delta from fetal calf thymus. J Biol Chem 1986, 261, 12310-12316
  25. Waga S, Hannon GJ, Beach D, Stillman B. The p21 inhibitor of cyclin-dependent kinases controls DNA replication by interaction with PCNA. Nature 1994, 369, 574-578 https://doi.org/10.1038/369574a0
  26. Wenz F, Azzam El, Little JB. The response of proliferating cell nuclear antigen to ionizing radiation in human lymphoblastoid cell lines is dependent on p53. Radiat Res 1998, 149, 32-40 https://doi.org/10.2307/3579679
  27. Xiong Y, Hannon GJ, Zhang H, Casso D, Kobayashi R, Beach D. p21 is a universal inhibitor of cyclin kinases. Nature 1993, 366, 701-704 https://doi.org/10.1038/366701a0
  28. Xiong Y, Zhang H, Beach D. D type cyclins associate with multiple protein kinases and the DNA replication and repair factor PCNA. Cell 1992, 71, 505-514 https://doi.org/10.1016/0092-8674(92)90518-H
  29. Zhan Q, Fan S, Bae I, Guillout C, Liebermann DA, O'Connor PM, Fornace AJ Jr. Induction of bax by genotoxic stress in human cells correlates with normal p53 status and apoptosis. Oncogene 1994, 9, 3743-3751
  30. Zhang H, Xiong Y, Beach D. Proliferating cell nuclear antigen and p21 are components of multiple cell cycle kinase complexes. Mol Biol Cell 1993, 4, 897-906 https://doi.org/10.1091/mbc.4.9.897