생명과학회지 (Journal of Life Science)

  • 제21권10호
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  • Pages.1494-1499
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  • 2011
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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β-lapachone에 의한 A549 인체폐암세포의 apoptosis 유도와 cyclooxygenase-2 활성 저하

β-Lapachone-Induced Apoptosis is Associated with Inhibition of Cyclooxygenase-2 Activity in Human Lung Cancer A549 Cells

  • 최영현 (동의대학교 한의과대학 생화학교실, 대학원 바이오물질제어학과(BK21 Program) 및 블루바이오소재개발센터)
  • Choi, Yung-Hyun (Department of Biochemistry, Dongeui University College of Oriental Medicine, Department of Biomaterial Control (BK21 program) and Blue-Bio Industry Regional Innovation Center, Dongeui University)
  • 투고 : 2011.08.12
  • 심사 : 2011.10.17
  • 발행 : 2011.10.31
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초록

${\beta}$-lapachone은 남미에 자생하는 lapacho 나무(Tabeuia avellanedae)의 수액에 함유된 quinone계열의 일종으로 많은 인체암세포에서 apoptosis를 유발하는 것으로 알려져 있다. 본 연구는 A549 인체폐암세포를 대상으로 ${\beta}$-lapachone에 의한 apoptosis 유발 과정에서 나타나는 또 다른 현상들을 조사하기 위하여 실시되었다. ${\beta}$-lapachone이 처리된 A549 세포는 처리 농도의 증가에 따라 생존율이 감소되었으며, 이는 apoptosis 유발과 연관이 있음을 MTT assay와 flow cytometry 분석을 통하여 확인하였다. ${\beta}$-lapachone에 의한 A549 세포의 증식억제는 종양억제유전자 p53과 cyclin-dependent kinase 저해제인 p21의 발현을 전사 및 번역 수준에서 증가시켰으며, p53 단백질의 인산화 증가와 연관성이 있었다. 또한 ${\beta}$-lapachone은 caspase-3과 -9를 활성화시켰으며, 활성화된 caspase-3의 기질 단백질들[poly(ADP-ribose) polymerase, ${\beta}$-catenin 및 phospholipase C-$\gamma$1]의 단편화를 유도하였다. 아울러 ${\beta}$-lapachone은 cyclooxygenase (COX)-2의 mRNA 및 단백질의 발현을 억제하였으나 COX-1의 발현에는 영향을 미치지 않았으며, ${\beta}$-lapachone에 의한 COX-2의 발현억제는 prostaglandin E2의 생성 저하에 관련이 있었다. 본 연구의 결과는 ${\beta}$-lapachone의 항암활성 기전의 이해와 더불어 ${\beta}$-lapachone이 폐암세포에서 강력한 항암활성이 있음을 보여 주는 것이다.

${\beta}$-lapachone, a quinone of lapachol extracted from the bark of the lapacho tree, has been found to induce apoptosis in various human cancer cells. In the present study, we investigated further possible mechanisms by which ${\beta}$-lapachone exerts its pro-apoptotic action in cultured human lung cancer A549 cells. ${\beta}$-lapachone treatment resulted in inhibition of growth and induction of apoptosis in a concentration-dependent manner, as determined by MTT assay and flow cytometry analysis. The induction of apoptosis by ${\beta}$-lapachone was associated with up-regulation of the expression of p53 and p21 in both transcriptional and translational levels, and the phosphorylation of p53. In addition, ${\beta}$-lapachone activated caspase-3 and -9, and induced degradation of caspase-3 target proteins such as poly (ADP-ribose) polymerase (PARP) and ${\beta}$-catenin. Furthermore, ${\beta}$-lapachone treatment caused a progressive decrease in the expression levels of cyclooxygenase (COX)-2 without significant changes in the levels of COX-1, which was correlated with a decrease in prostaglandin E2 synthesis. Taken together, these results indicated that ${\beta}$-lapachone may have therapeutic potential in human lung cancer treatment.

키워드

참고문헌

  1. Anderson, W. F., A. Umar, J. L. Viner, and E. T. Hawk. 2002. The role of cyclooxygenase inhibitors in cancer prevention. Curr. Pharm. 8, 1035-1062. https://doi.org/10.2174/1381612023394935
  2. Choi, Y. H., H. S. Kang, and M. A. Yoo. 2002. Suppression of Human Prostate Cancer Cell Growth by $\beta$-lapachone via Inhibition of pRB Phosphorylation and Induction of Cdk inhibitor p21. J. Biochem. Mol. Biol. 36, 223-229.
  3. Choi, Y. H., M. J. Kim, S. Y. Lee, Y. N. Lee, G. Y. Chi, H. S. Eom, N. D. Kim, and B. T. Choi. 2002. Phosphorylation of p53, induction of Bax and activation of caspases during $\beta$-lapachone-mediated apoptosis in human prostate epithelial cells. Int. J. Oncol. 21, 1293-1299.
  4. Don, M. J., Y. H. Chang, K. K. Chen, L. K. Ho, and Y. P. Chau. 2001. Induction of CDK inhibitors (p21(WAF1) and p27(Kip1)) and Bak in the $\beta$-lapachone-induced apoptosis of human prostate cancer cells. Mol. Pharmacol. 59, 784-794.
  5. Fujita, H., K. Koshida, E. T. Keller, Y. Takahashi, T. Yoshimito, M. Namiki, and A. Mizokami. 2002. Cyclooxygenase-2 promotes prostate cancer progression. Prostate 53, 232-240. https://doi.org/10.1002/pros.10152
  6. Gately, S. and R. Kerbel. 2003. Therapeutic potential of selective cyclooxygenase-2 inhibitors in the management of tumor angiogenesis. Prog. Exp. Tumor. Res. 37, 179-192. https://doi.org/10.1159/000071373
  7. Huang, L., and A. B. Pardee. 1999. $\beta$-lapachone induces cell cycle arrest and apoptosis in human colon cancer cells. Mol. Med. 5, 711-720.
  8. Hueber, A. P. Esser, K. Heimann, N. Kociok, S. Winter, and M. Weller. 1998. The topoisomerase I inhibitors, camptothecin and $\beta$-lapachone, induce apoptosis of human retinal pigment epithelial cells. Exp. Eye Res. 67, 525-530. https://doi.org/10.1006/exer.1998.0544
  9. Lee, J. H., J. Cheong, Y. M. Park, and Y. H. Choi. 2005. Down-regulation of cyclooxygenase-2 and telomerase activity by $\beta$-lapachone in human prostate carcinoma cells. Pharmacol. Res. 51, 553-560. https://doi.org/10.1016/j.phrs.2005.02.004
  10. Lee, J. I., D. Y. Choi, H. S. Chung, H. G. Seo, H. J. Woo, B. T. Choi, and Y. H. Choi. 2006. $\beta$-lapachone induces growth inhibition and apoptosis in bladder cancer cells by modulation of Bcl-2 family and activation of caspases. Exp. Oncol. 28, 30-35.
  11. Li, C. J., L. Averboukh, and A. B. Pardee. 1993. $\beta$-Lapachone, a novel DNA topoisomerase I inhibitor with a mode of action different from camptothecin. J. Biol. Chem. 268, 22463-22468.
  12. Meng, R. D. and W. S. El-Deiry. 2001. p53-independent upregulation of KILLER/DR5 TRAIL receptor expression by glucocorticoids and interferon-$\gamma$. Exp. Cell Res. 262, 154-169. https://doi.org/10.1006/excr.2000.5073
  13. Moon, D. O., Y.H. Choi, N. D. Kim, Y.M. Park, and G. Y. Kim. 2007. Anti-inflammatory effects of $\beta$-lapachone in lipopolysaccharide-stimulated BV2 microglia. Int. Immunopharmacol. 7, 506-514. https://doi.org/10.1016/j.intimp.2006.12.006
  14. Noble, J. R., K. E. Willetts, W. E. Mercer, and R. R. Reddel. 1992. Effects of exogenous wild-type p53 on a human lung carcinoma cell line with endogenous wild-type p53. Exp. Cell Res. 203, 297-304. https://doi.org/10.1016/0014-4827(92)90002-P
  15. Pink, J. J., S. M. Planchon, C. Tagliarino, M. E. Varnes, D. Siegel, and D. A. Boothman. 2000. NAD(P)H:Quinone oxidoreductase activity is the principal determinant of $\beta$ -lapachone cytotoxicity. J. Biol. Chem. 275, 5416-5424. https://doi.org/10.1074/jbc.275.8.5416
  16. Planchon, S. M., J. J. Pink, C. Tagliarino, W. G. Bornmann, M. E. Varnes, and D. A. Boothman. 2001. $\beta$-lapachone-induced apoptosis in human prostate cancer cells: involvement of NQO1/xip3. Exp. Cell Res. 267, 95-106. https://doi.org/10.1006/excr.2001.5234
  17. Planchon, S. M., S. Wuerzberger, B. Frydman, D. T. Witiak, P. Hutson, D. R. Church, G. Wilding, and D. A. Boothman. 1995. $\beta$-lapachone-mediated apoptosis in human promyelocytic leukemia (HL-60) and human prostate cancer cells: a p53-independent response. Cancer Res. 55, 3706-3711.
  18. Planchon, S. M., S. M. Wuerzberger-Davis, J. J. Pink, K. A. Robertson, W. G. Bornmann, and D. A. Boothman. Bcl-2 protects against $\beta$-lapachone-mediated caspase 3 activation and apoptosis in human myeloid leukemia (HL-60) cells. Oncol. Rep. 6, 485-492.
  19. Schaffner-Sabba, K., K. H. Schmidt-Ruppin, W. Wehrli, A. R. Schuerch, and J.W. Wasley. 1984. $\beta$-lapachone: synthesis of derivatives and activities in tumor models. J. Med. Chem. 27, 990-994. https://doi.org/10.1021/jm00374a010
  20. Weller, M., J. Rieger, C. Grimmel, E.G. Van Meir, N. De Tribolet, S. Krajewski, J. C. Reed, A. von Deimling, and J. Dichgans. 1998. Predicting chemoresistance in human malignant glioma cells: the role of molecular genetic analyses. Int. J. Cancer 79, 640-644. https://doi.org/10.1002/(SICI)1097-0215(19981218)79:6<640::AID-IJC15>3.0.CO;2-Z
  21. Weller, M., S. Winter, C. Schmidt, P. Esser, A. Fontana, J. Dichgans, and P. Groscurth. 1997. Topoisomerase-I inhibitors for human malignant glioma: differential modulation of p53, p21, bax and bcl-2 expression and of CD95-mediated apoptosis by camptothecin and $\beta$ -lapachone. Int. J. Cancer 73, 707-714. https://doi.org/10.1002/(SICI)1097-0215(19971127)73:5<707::AID-IJC16>3.0.CO;2-2
  22. Wuerzberger, S. M., J. J. Pink, S. M. Planchon, K. L. Byers, W. G. Bornmann, and D. A. Boothman. 1998. Induction of apoptosis in MCF-7:WS8 breast cancer cells by $\beta$-lapachone. Cancer Res. 58, 1876-1885.