Journal of the Korean Society of Food Science and Nutrition (한국식품영양과학회지)

The Korean Society of Food Science and Nutrition (한국식품영양과학회)
권호 표지
DOI QR코드

DOI QR Code

Induction of Apoptosis in HT-29 Human Colon Cancer Cells by the Pepper Component Piperine

후추의 주요 성분인 Piperine의 대장암세포 세포사멸 유도 효과

  • Kim, Eun-Ji (Center for Efficacy Assessment and Development of Functional Foods and Drugs, Hallym University) ;
  • Park, Hee-Sook (Dept. of Food Science and Nutrition, Hallym University) ;
  • Shin, Min-Jeong (Dept. of Food Science and Nutrition, Hallym University) ;
  • Shin, Hyun-Kyung (Dept. of Food Science and Nutrition, Hallym University) ;
  • YoonPark, Jung-Han (Dept. of Food Science and Nutrition, Hallym University)
  • 김은지 (한림대학교 식의약품의 효능평가 및 기능성소재개발센터) ;
  • 박희숙 (한림대학교 식품영양학과) ;
  • 신민정 (한림대학교 식품영양학과) ;
  • 신현경 (한림대학교 식품영양학과) ;
  • 윤정한 (한림대학교 식품영양학과)
  • Published : 2009.04.30
Download PDF
⟨ Previous Next ⟩

Abstract

Piperine is an alkaloid-amine found in pepper and has been reported to have anticarcinogenic properties. To explore the possibility that piperine has cancer chemopreventive and chemotherapeutic effects in colon cancer, we examined whether piperine inhibits the growth of HT-29 human colon cancer cells and investigated the mechanisms for this effect. Cells were cultured with various concentrations ($0{\sim}40{\mu}M$) of piperine. Piperine decreased the cell viability and induced apoptosis of HT-29 cells. Western blot analysis of total cell lysates revealed that piperine decreases the protein levels of Bcl-2, Mcl-1, and intact Bid but increases Bik levels. Piperine increased the percentage of cells with depolarized mitochondrial membrane, and the release of cytochrome c into cytoplasm. Piperine induced the cleavage of poly (ADP-ribose) polymerase and caspases 8, 9, 7, and 3 and increased the Fas levels. In addition, piperine significantly decreased the protein levels of survivin. The present results indicate that piperine inhibits the growth of HT-29 colon cancer cells by the induction of apoptosis, which may be mediated by its ability to change the Bcl-2 family proteins, increase the activation of caspases, and decrease survivin levels. Overall, our findings suggest that piperine has cancer chemotherapeutic effects in colon cancer.

후추의 주요 성분인 piperine은 다양한 생리활성을 나타내고 있으며, 특히 암예방 효과가 있는 것으로 생각되고 있다. 본 연구에서는 piperine의 항암 효과를 밝히기 위해 piperine이 인간의 대장에서 유래한 암세포인 HT-29 세포의 증식에 미치는 영향과 작용 기전을 연구하였다. Piperine을 HT-29 세포 배양액에 여러 농도($0{\sim}40{\mu}M$)로 첨가하여 세포를 배양한 경우 piperine 처리 농도가 증가할수록 세포의 증식이 감소하였고, 세포사멸이 증가하였다. 이는 piperine이 HT-29 세포의 세포사멸을 유도하여 세포 증식을 억제함을 제시한다. Piperine의 세포사멸 기전을 조사하기 위해 세포사멸 조절인자의 변화를 조사하였다. Piperine에 의해 anti-apoptotic Bcl-2 family 단백질인 Bcl-2와 Mcl-1 단백질 수준은 감소하였고, BH3-only 단백질인 Bid 단백질 수준은 감소하였으나, Bik 단백질 수준은 증가하였다. 또한 piperine에 의해 미토콘드리아 막의 투과성이 증가하였고, cytochrome c의 세포질로의 방출이 증가하였다. 또한 piperine 처리에 의해 caspase의 활성형인 cleaved caspase-8, -9, -7, -3 단백질 수준이 증가하였고, PARP의 불활성형인 cleaved PARP 수준이 증가하였다. Caspase의 활성을 저해하는 세포사멸억제단백질 중의 하나인 survivin 단백질 발현이 piperine에 의해 감소하였다. 이 결과로부터 대장암세포인 HT-29 세포에서 piperine이 Bcl-2 family 단백질 발현 변화를 초래하여 미토콘드리아 막 투과성 증가시키고 cytochrome c 방출을 증가시키고, caspase 활성을 증가시키고 survivin 단백질 발현을 억제하여 세포사멸을 유도하여 항암 효과를 나타냄을 알 수 있다. 본 연구는 piperine이 대장암에 강한 항암 효과가 있음을 밝혔으나 향후 암예방 및 암치료제로서 piperine을 활용하기 위해서는 동물실험 및 임상실험 등 다양한 추가 실험이 필요할 것으로 보인다.

Keywords

References

  1. Jemal A, Siegel R, Ward E, Murray T, Xu J, Thun MJ. 2007. Cancer statistics, 2007. CA Cancer J Clin 57: 43-66 https://doi.org/10.3322/canjclin.57.1.43
  2. Kim WS, Lee RA, Hwang DY, Hong YJ, Hong SI. 2004. Histoculture drug response assay in colorectal cancer specimen. J Korean Surg Soc 66: 109-115
  3. Maroun JA, Anthony LB, Blais N, Burkes R, Dowden SD, Dranitsaris G, Samson B, Shah A, Thirlwell MP, Vincent MD, Wong R. 2007. Prevention and management of chemptherapy-induced diarrhea in patients with colorectal cancer: a consensus statement by the canadian working group on chemotherapy-induced diarrhea. Curr Oncol 14: 13-20 https://doi.org/10.3747/co.2007.96
  4. de Gramount A, Figer A, Seymour M, Homerin M, Hmissi A, Cassidy J, Boni C, Cortes-Funes H, Cervantes A, Freyer G, Papamichael D, Le Bail N, Louvet C, Hendler D, de Braud F, Wilson C, Morvan F, Bonetti A. 2000. Leucovorin and fluorouracil with or without oxaliplatin as first-line treatmet in advanced colorectal cancer. J Clin Oncol 18: 2938-2947 https://doi.org/10.1200/JCO.2000.18.16.2938
  5. Parmar VS, Jain SC, Bisht KS, Jain R, Taneja P, Jha A, Tyagi OD, Prasad AK, Wengel J, Olsen CE, Boll PE. 1997. Phytochemistry of the genus Piper. Phytochemistry 46: 597-673 https://doi.org/10.1016/S0031-9422(97)00328-2
  6. Mittal R, Gupta RI. 2000. In vitro antioxidant activity of piperine. Methods Find Exp Clin Pharmacol 22: 271-274 https://doi.org/10.1358/mf.2000.22.5.796644
  7. Singh J, Reen RK, Wiebel FJ. 1994. Piperine, a major ingredient of black and long peppers, protects against AFB1-induced cytotoxicity and micronucliei formation in H4IIEC3 rat hepatoma cells. Cancer Lett 86: 195-200 https://doi.org/10.1016/0304-3835(94)90078-7
  8. Lee CS, Han ES, Kim YK. 2006. Piperine inhibition of 1-methyl-4-phenylpyridinium-induced mitochondrial dysfunction and cell death in PC12 cells. Eur J Pharmacol 537: 37-44 https://doi.org/10.1016/j.ejphar.2006.03.026
  9. Pradeep CR, Kuttan G. 2004. Piperine is a potent inhibitor of nuclear factor-kappaB (NF-kappaB), s-Fos, CREP, ATF-2 and proinflammatory cytokine gene expression in B16F-10 melanoma cells. Int Immunopharmacol 4: 1795-1803 https://doi.org/10.1016/j.intimp.2004.08.005
  10. Selvendiran K, Banu SM, Sakthisekaran D. 2004. Protective effect of piperine on benzo(a)pyrene-induced lung carcinogenesis in Swiss albino mice. Clin Chim Acta 350: 73-78 https://doi.org/10.1016/j.cccn.2004.07.004
  11. Selvendiran K, Singh JPV, Sakthisekaran D. 2006. In vivo effect of piperine on serum and tissue glycoprotein levels in benzo(a)pyrene induced lung carcinogenesis in Swiss albino mice. Pulm Pharmacol Ther 19: 107-111 https://doi.org/10.1016/j.pupt.2005.04.002
  12. Pradeep CR, Kuttan G. 2002. Effect of piperine on the inhibition of lung metastasis induced B16F-10 melanoma cells in mice. Clin Exp Metastasis 19: 703-708 https://doi.org/10.1023/A:1021398601388
  13. Bezerra DP, Castro FO, Alves AP, Pessoa C, Moraes MO, Silveira ER, Lima MA, Elmiro FJ, Costa-Lotufo LV. 2006. In vivo growth-inhibition of Sarcoma 180 by piplartine and piperine, two alkaloid amides from piper. Braz J Med Biol Res 39: 801-807 https://doi.org/10.1590/S0100-879X2006000600014
  14. Duessel S, Heuertz RM, Ezekiel UR. 2008. Growth inhibition of human colon cancer cells by plant compounds. Clin Lab Sci 21: 151-157
  15. Hengarther MO. 2000. The biochemistry of apoptosis. Nature 407: 770-776 https://doi.org/10.1038/35037710
  16. Jin Z, El-Deiry WS. 2005. Overview of cell death signaling pathways. Cancer Biol Ther 4: 139-163 https://doi.org/10.4161/cbt.4.2.1508
  17. Yao J, Jiang Z, Duan W, Huanq J, Zhanq L, He L, Li F, Xiao Y, Shu B, Lin C. 2008. Involvement of mitochondrial pathway in triptolide-induced cytotoxicity in human normal liver L-02 cells. Biol Pharm Bull 31: 592-597 https://doi.org/10.1248/bpb.31.592
  18. Ambrosini G, Adida C, Altieri DA. 1997. A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma. Nat Med 3: 917-921 https://doi.org/10.1038/nm0897-917
  19. Altieri DC, Marchisio PC. 1999. Survivin apoptosis: an interloper between cell death and cell proliferation in cancer. Lab Invest 79: 1327-1333
  20. Reed JC. 2004. Apoptosis mechanisms: implications for cancer drug discovery. Oncology 18: 10-20 https://doi.org/10.1159/000218690
  21. Frankfurt OS, Krishan A. 2003. Apoptosis-based drug screening and detection of selective toxicity to cancer cells. Anticancer Drugs 14: 555-561 https://doi.org/10.1097/00001813-200308000-00008
  22. Lowe SW, Lin AW. 2000. Apoptosis in cancer. Carcinogenesis 21: 485-495 https://doi.org/10.1093/carcin/21.3.485
  23. Kim EJ, Park SY, Hong J, Shin M, Lim SS, Shin HK, Park JHY. 2007. Inhibitory effect of the methanolic extract of Symphyocladia latiuscula on the growth of HT-29 human colon cancer cells. J Korean Soc Food Sci Nutr 36: 431-438 https://doi.org/10.3746/jkfn.2007.36.4.431
  24. Kim EJ, Park H, Lim SS, Kim JS, Shin HK, Park JHY. 2008. Effect of the hexane extracts of Saussure lappa on the growth of HT-29 human colon cancer cells. Korean J Food Sci Technol 40: 207-214
  25. Kim EJ, Park SY, Shin HK, Kwon DY, Surh YJ, Park JHY. 2007. Activation of caspase-8 contributes to 3,3'-diindolymethane-induced apoptosis in colon cancer cells. J Nutr 137: 1-6
  26. Kim EJ, Lee YJ, Shin HK, Park JHY. 2006. A study on the mechanisms by which the aqueous extract of Inonotus obliquus induces apoptosis and inhibits proliferation in HT-29 human colon cancer cells. J Korean Soc Food Sci Nutr 35: 516-523 https://doi.org/10.3746/jkfn.2006.35.5.516
  27. Denizot F, Lang R. 1986. Rapid colorimetric assay for cell growth and survival modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods 89: 271-277 https://doi.org/10.1016/0022-1759(86)90368-6
  28. Kim EJ, Shin HK, Cho JS, Lee SK, Won MH, Kim HW, Park JHY. 2006. Trans-10, cis-12 conjugated linoleic acid inhibits the G1-S cell cycle progression in DU145 human prostate carcinoma cells. J Med Food 9: 293-299 https://doi.org/10.1089/jmf.2006.9.293
  29. Jung JI, Lim SS, Choi HJ, Cho HJ, Shin HK, Kim EJ, Chung WY, Park KK, Park JH. 2006. Isoliquiritigenin induces apoptosis by depolarizing mitochondrial membranes in prostate cancer cells. J Nutr Biochem 17: 689-696 https://doi.org/10.1016/j.jnutbio.2005.11.006
  30. Eguchi Y, Srinivasan A, Tomaselli KJ, Shimizu S, Tsujimoto Y. 1999. ATP-dependent steps in apoptotic signal transduction. Cancer Res 59: 2174-2181
  31. Sunila ES, Kuttan G. 2004. Immunomodulatory and antitumor activity of Pier longum Linn. and piperine. J Ethnopharmacol 90: 339-346 https://doi.org/10.1016/j.jep.2003.10.016
  32. Yu Z, Li W. 2006. Induction of apoptosis by puerarin in colon cancer HT-29 cells. Cancer Lett 238: 53-60 https://doi.org/10.1016/j.canlet.2005.06.022
  33. Luo X, Budihardjo I, Zou H, Slaughter C, Wang X. 1998. Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell 94: 481-490 https://doi.org/10.1016/S0092-8674(00)81589-5
  34. Oliver FJ, de la Rubia G, Rolli V, Ruiz-Ruiz MC, de Murcia G, Murcia JM. 1998. Importance of poly(ADP-ribose) polymerase and its cleavage in apoptosis. Lesson from an uncleavable mutant. J Biol Chem 273: 33533-33539 https://doi.org/10.1074/jbc.273.50.33533
  35. Budihardjo I, Oliver H, Lutter M, Wang X. 1999. Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol 15: 269-290 https://doi.org/10.1146/annurev.cellbio.15.1.269
  36. Ashkenazi A, Dixit VM. 1998. Death receptor: signaling and modulation. Science 281: 1305-1308 https://doi.org/10.1126/science.281.5381.1305
  37. Nijhawan LP, Budihardjo D, Srinivasula I, Ahmad SM, Alnemri M, Wang X. 1997. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91: 479-489 https://doi.org/10.1016/S0092-8674(00)80434-1
  38. Baker SJ, Reddy EP. 1998. Modulation of life and death by the TNF receptor superfamily. Oncogene 17: 3261-3270 https://doi.org/10.1038/sj.onc.1202568

Cited by

  1. 삼지구엽초의 인체 대장암세포 HCT116 사멸 효과 vol.18, pp.2, 2014, https://doi.org/10.13050/foodengprog.2014.18.2.154
  2. Resveratrol Induces Apoptosis in Primary Human Prostate Cancer Cells vol.39, pp.8, 2010, https://doi.org/10.3746/jkfn.2010.39.8.1119
  3. Piperine-A Major Principle of Black Pepper: A Review of Its Bioactivity and Studies vol.9, pp.20, 2009, https://doi.org/10.3390/app9204270