Radiation Oncology Journal

  • 제29권2호
  • /
  • Pages.83-90
  • /
  • 2011
  • /
  • 2234-1900(pISSN)
  • /
  • 2234-3156(eISSN)
대한방사선종양학회 (The Korean Society for Radiation Oncology)
권호 표지
DOI QR코드

DOI QR Code

마우스를 이용한 생체내 실험에서의 플라보피리돌의 방사선민감화 효과

In Vitro Radiosensitization of Flavopiridol Did Not Translated into In Vivo Radiosensitization

  • 김수지 (가톨릭대학교 의과대학 방사선종양학교실)
  • Kim, Su-Zy (Department of Radiation Oncology, The Catholic University of Korea School of Medicine)
  • 투고 : 2011.04.05
  • 심사 : 2011.05.17
  • 발행 : 2011.06.30
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

목 적: 이전의 암세포주를 이용한 실험실내 연구에서 플라보피리돌은 암세포의 방사선에 의한 아포토시스를 증가시키는 것을 알 수 있었다. 이번 연구에서는 마우스를 이용한 생체내 실험에서 플라보피리돌의 방사선민감화 효과를 알아보고자 하였다. 대상 및 방법: 마우스 유방암 세포주인 EMT-6를 Balb/c 마우스에 피하주사하여 종양을 만든 후 플라보피리돌 단독 치료군, 방사선 단독 치료군, 방사선과 플라보피리돌 병합 치료군 및 대조군으로 나누어 종양의 성장 속도를 비교 하였다. 플라보피리톨은 2.5 mg/kg을 하루 2회 복강내에 주사하였고, 방사선은 1일 1회, 회당 4 Gy를 조사하여 총 28 Gy를 조사하였다. 각 치료군에서의 종양 성장 곡선을 구하여 비교하였다. 마우스로부터 채취한 종양으로 파라핀 절편을 만틀어 TUNEL 및 면역조직화학염색을 시행하였다. 결 과: 종양 성장을 비교하였을 때 대조군보다 방사선 단독 치료군과 방사선과 플라보피리돌 병합 치료군에서 종양 성장이 지연되는 것을 볼 수 있었다. 그러나 대조군과 플라보피리돌 단독 치료군 사이에서는 종양 성장에 차이가 없었고, 방사선 단독 치료군과 방사선과 플라보피리돌 병합 치료군 사이에서도 차이가 없었다. TUNEL 염색으로 아포토시스율을 비교했을 때 각 치료군 사이에 차이가 없었으며, 면역조직화학염색으로 Ku70 발현을 비교했을 때에도 각 치료군 사이에 차이가 없었다. 결 론: 플라보피리돌은 마우스 유방암 모델에서 방사선민감화 효과를 나타내지 않았다.

Purpose: Flavopiridol enhanced radiation-induced apoptosis of cancer cells in our previous in vitro study. The purpose of this study was to assess if flavopiridol could enhance the radioresponse of mouse mammary tumors in vivo. Materials and Methods: Balb/c mice bearing EMT-6 murine mammary carcinoma were treated with flavopiridol only, radiation only, or both for 7 days. Flavopiridol was administered 2.5 mg/kg twice a day intraperitoneally (IP). Radiation was delivered at a 4 Gy/fraction at 24-h intervals for a total dose of 28 Gy. Tumor volume was measured and compared among the different treatment groups to evaluate the in vivo radiosensitizing effect of flavopiridol. Tumors were removed from the mice 20 days after treatment, and TUNEL and Immunohistochemical stainings were performed. Results: Significant tumor growth delay was observed in the radiation only and combined treatment groups, when compared with the control group. However, there was no significant difference between the tumor growth curves of the control and flavopiridol only group or between the radiation only and combination treatment group. Apoptotic cells of different treatment groups were detected by terminal deoxynucleotidyl transferase-medicated nick end labeling (TUNEL) staining. The expressions of Ku70 in tumor tissues from the different groups were analyzed by immunohistochemistry. Similarly, no significant difference was found between the apoptotic rate or Ku70 expression among the different treatment groups. Conclusion: Flavopiridol did not show evidence of enhancing the radioresponse of mouse mammary tumors in this study.

키워드

참고문헌

  1. Senderowicz AM. Flavopiridol: the first cyclin-dependent kinase inhibitor in human clinical trials. Invest New Drugs 1999;17:313-320 https://doi.org/10.1023/A:1006353008903
  2. Senderowicz AM, Sausville EA. Preclinical and clinical development of cyclin-dependent kinase modulators. J Natl Cancer Inst 2000;92:376-387 https://doi.org/10.1093/jnci/92.5.376
  3. Worland PJ, Kaur G, Stetler-Stevenson M, Sebers S, Sartor O, Sausville EA. Alteration of the phosphorylation state of p34cdc2 kinase by the flavone L86-8275 in breast carcinoma cells: correlation with decreased H1 kinase activity. Biochem Pharmacol 1993;46:1831-1840 https://doi.org/10.1016/0006-2952(93)90590-S
  4. Sedlacek HH. Mechanisms of action of flavopiridol. Crit Rev Oncol Hematol 2001;38:139-170 https://doi.org/10.1016/S1040-8428(00)00124-4
  5. Patel V, Senderowicz AM, Pinto D Jr, et al. Flavopiridol, a novel cyclin-dependent kinase inhibitor, suppresses the growth of head and neck squamous cell carcinomas by inducing apoptosis. J Clin Invest 1998;102:1674-1681 https://doi.org/10.1172/JCI3661
  6. Melillo G, Sausville EA, Cloud K, Lahusen T, Varesio L, Senderowicz AM. Flavopiridol, a protein kinase inhibitor, down-regulates hypoxic induction of vascular endothelial growth factor expression in human monocytes. Cancer Res 1999;59:5433-5437
  7. Bible KC, Kaufmann SH. Cytotoxic synergy between flavopiridol (NSC 649890, L86-8275) and various antineoplastic agents: the importance of sequence of administration. Cancer Res 1997;57:3375-3380
  8. Motwani M, Delohery TM, Schwartz GK. Sequential dependent enhancement of caspase activation and apoptosis by flavopiridol on paclitaxel-treated human gastric and breast cancer cells. Clin Cancer Res 1999;5:1876-1883
  9. Raju U, Nakata E, Mason KA, Ang KK, Milas L. Flavopiridol, a cyclin-dependent kinase inhibitor, enhances radiosensitivity of ovarian carcinoma cells. Cancer Res 2003;63:3263-3267
  10. Jung C, Motwani M, Kortmansky J, et al. The cyclin-dependent kinase inhibitor flavopiridol potentiates gamma-irradiation-induced apoptosis in colon and gastric cancer cells. Clin Cancer Res 2003;9:6052-6061
  11. Newcomb EW, Lymberis SC, Lukyanov Y, et al. Radiation sensitivity of GL261 murine glioma model and enhanced radiation response by flavopiridol. Cell Cycle 2006;5:93-99 https://doi.org/10.4161/cc.5.1.2271
  12. Kim S, Wu HG, Shin JH, Park HJ, Kim IA, Kim IH. Enhancement of radiation effects by flavopiridol in uterine cervix cancer cells. Cancer Res Treat 2005;37:191-195 https://doi.org/10.4143/crt.2005.37.3.191
  13. Kim S, Kwon EK, Lee SH, Park HJ, Wu HG. Effect of flavopiridol on radiation-induced apoptosis of human laryngeal and lung cancer cells. J Korean Soc Ther Radiol Oncol 2007;25:227-232
  14. Jeggo PA. Identification of genes involved in repair of DNA double-strand breaks in mammalian cells. Radiat Res 1998;150(5 Suppl):S80-S91 https://doi.org/10.2307/3579810
  15. Omori S, Takiguchi Y, Suda A, et al. Suppression of a DNA double-strand break repair gene, Ku70, increases radio-and chemosensitivity in a human lung carcinoma cell line. DNA Repair (Amst) 2002;1:299-310 https://doi.org/10.1016/S1568-7864(02)00006-X
  16. Mason KA, Hunter NR, Raju U, et al. Flavopiridol increases therapeutic ratio of radiotherapy by preferentially enhancing tumor radioresponse. Int J Radiat Oncol Biol Phys 2004;59:1181-1189 https://doi.org/10.1016/j.ijrobp.2004.03.003
  17. MacLachlan TK, Sang N, Giordano A. Cyclins, cyclindependent kinases and cdk inhibitors: implications in cell cycle control and cancer. Crit Rev Eukaryot Gene Expr 1995;5:127-156 https://doi.org/10.1615/CritRevEukarGeneExpr.v5.i2.20
  18. Patel V, Jakus J, Harris CM, Ensley JF, Robbins KC, Yeudall WA. Altered expression and activity of G1/S cyclins and cyclin-dependent kinases characterize squamous cell carcinomas of the head and neck. Int J Cancer 1997;73:551-555 https://doi.org/10.1002/(SICI)1097-0215(19971114)73:4<551::AID-IJC16>3.0.CO;2-C
  19. Losiewicz MD, Carlson BA, Kaur G, Sausville EA, Worland PJ. Potent inhibition of CDC2 kinase activity by the flavonoid L86-8275. Biochem Biophys Res Commun 1994;201:589-595 https://doi.org/10.1006/bbrc.1994.1742
  20. Raju U, Ariga H, Koto M, et al. Improvement of esophageal adenocarcinoma cell and xenograft responses to radiation by targeting cyclin-dependent kinases. Radiother Oncol 2006;80:185-191 https://doi.org/10.1016/j.radonc.2006.07.027
  21. Willingham MC. Cytochemical methods for the detection of apoptosis. J Histochem Cytochem 1999;47:1101-1110 https://doi.org/10.1177/002215549904700901
  22. Gu Y, Jin S, Gao Y, Weaver DT, Alt FW. Ku70-deficient embryonic stem cells have increased ionizing radiosensitivity, defective DNA end-binding activity, and inability to support V(D)J recombination. Proc Natl Acad Sci USA 1997;94:8076-8081 https://doi.org/10.1073/pnas.94.15.8076
  23. Ouyang H, Nussenzweig A, Kurimasa A, et al. Ku70 is required for DNA repair but not for T cell antigen receptor gene recombination in vivo. J Exp Med 1997;186:921-929 https://doi.org/10.1084/jem.186.6.921
  24. Camphausen K, Brady KJ, Burgan WE, et al. Flavopiridol enhances human tumor cell radiosensitivity and prolongs expression of gammaH2AX foci. Mol Cancer Ther 2004;3:409-416
  25. Hara T, Omura-Minamisawa M, Kang Y, Cheng C, Inoue T. Flavopiridol potentiates the cytotoxic effects of radiation in radioresistant tumor cells in which p53 is mutated or Bcl-2 is overexpressed. Int J Radiat Oncol Biol Phys 2008;71:1485-1495 https://doi.org/10.1016/j.ijrobp.2008.03.039