, Volume 13, Issue 12, pp 1465–1478 | Cite as

In vitro and in vivo induction of apoptosis by capsaicin in pancreatic cancer cells is mediated through ROS generation and mitochondrial death pathway

  • Ruifen Zhang
  • Ian Humphreys
  • Ravi P. Sahu
  • Yan Shi
  • Sanjay K. SrivastavaEmail author
Original Paper


Pancreatic cancer is one of the most common invasive malignancies and the fourth leading cause of cancer related mortality in U.S., thus developing new strategies to control pancreatic cancer is an important mission. We investigated the mechanism of capsaicin, the major pungent ingredient of red-chili pepper, in inducing apoptosis in pancreatic cancer cells. Treatment of AsPC-1 and BxPC-3 cells with capsaicin resulted in a dose-dependent inhibition of cell-viability and induction of apoptosis which was associated with the generation of ROS and persistent disruption of mitochondrial membrane potential. These effects were significantly blocked when the cells were pretreated with a general antioxidant N-acetyl cysteine (NAC). Exposure of AsPC-1 and BxPC-3 cells to capsaicin was also associated with increased expression of Bax, down-regulation of bcl-2, survivin and significant release of cytochrome c and AIF in the cytosol. On the contrary, above-mentioned effects were not observed in the normal acinar cells in response to capsaicin-treatment. Capsaicin-treatment resulted in the activation of JNK and JNK inhibitor SP600125 afforded protection against capsaicin-induced apoptosis. Furthermore, capsaicin when given orally markedly suppressed the growth of AsPC-1 pancreatic tumor xenografts in athymic nude mice, without side effects. Tumors from capsaicin treated mice demonstrated increased apoptosis, which was related to the activation of JNK and increased cytosolic protein expression of Bax, cytochrome c, AIF and cleaved caspase-3, as compared with controls. Taken together, these results show that capsaicin is an effective inhibitor of in vitro and in vivo growth of pancreatic cancer cells. These findings provide the rationale for further clinical investigation of capsaicin against pancreatic cancer.


Capsaicin Pancreatic cancer ROS Bax Cytochrome c Nude mice Xenograft 



This investigation was supported in part by USPHS RO1 grant CA106953 (to SKS) awarded by the National Cancer Institute. Funds from Texas Tech University Health Sciences Center, School of Pharmacy and Hillman Foundation (to SKS) are also acknowledged. The authors thank Dr. Massimo Trucco, University of Pittsburgh for providing acinar cells isolated from normal human pancreas and Shephalie Lahiri for technical assistance.


  1. 1.
    Polednak AP (1994) Projected numbers of cancers diagnosed in the US elderly population, 1990 through 2030. Am J Public Health 84:1313–1316PubMedGoogle Scholar
  2. 2.
    Niederhuber JE, Brennan MF, Menck HR (1995) The National Cancer Data Base Report on pancreatic cancer. Cancer 76:1617–1677. doi:10.1002/1097-0142(19951101)76:9<1671::AID-CNCR2820760926>3.0.CO;2-RCrossRefGoogle Scholar
  3. 3.
    DiMagno EP, Reber HA, Tempero MA (1999) AGA technical review on the epidemiology, diagnosis and treatment of pancreatic ductal adenocarcinoma: American Gastroenterological Association. Gastroenterology 117:1464–1484. doi: 10.1016/S0016-5085(99)70298-2 PubMedCrossRefGoogle Scholar
  4. 4.
    Li D (2001) Molecular epidemiology of pancreatic cancer. Cancer J 7:259–265PubMedGoogle Scholar
  5. 5.
    Ghaneh P, Kawesha A, Evans JD, Neoptolemos JP (2002) Molecular prognostic markers in pancreatic cancer. J Hepatobiliary Pancreat Surg 9:1–11. doi: 10.1007/s005340200000 PubMedCrossRefGoogle Scholar
  6. 6.
    DiGiuseppe JA, Yeo CJ, Hruban RH (1996) Molecular biology and the diagnosis and treatment of adenocarcinoma of the pancreas. Adv Anat Pathol 3:139–155Google Scholar
  7. 7.
    Schmid RM (2002) Acinar-to-ductal metaplasia in pancreatic cancer development. J Clin Invest 109:1403–1404PubMedGoogle Scholar
  8. 8.
    Banerjee S, Zhang Y, Wang Z, Che M, Chiao PJ, Abbruzzese JL, Sarkar FH (2007) In vitro and in vivo molecular evidence of genistein action in augmenting the efficacy of cisplatin in pancreatic cancer. Int J Cancer 120:906–917. doi: 10.1002/ijc.22332 PubMedCrossRefGoogle Scholar
  9. 9.
    Tyagi A, Agarwal R, Agarwal C (2003) Grape seed extract inhibits EGF-induced and constitutively activated mitogenic signaling but activates JNK in human prostate carcinoma DU145 cells: possible role in antiproliferation and apoptosis. Oncogene 22:1302–1316. doi: 10.1038/sj.onc.1206265 PubMedCrossRefGoogle Scholar
  10. 10.
    Shukla S, Mishra A, Fu P, MacLennan GT, Resnick MI, Gupta S (2005) Up-regulation of insulin-like growth factor binding protein-3 by apigenin leads to growth inhibition and apoptosis of 22Rv1 xenograft in athymic nude mice. FASEB J 19:2042–2044PubMedGoogle Scholar
  11. 11.
    Rao CV, Reddy BS, Steele VE, Wang C, Liu X, Ouyang N (2006) Nitric oxide releasing aspirin and indomethacin are potent inhibitors against colon cancer in azoxymethane-treated rats: effects on molecular targets. Mol Can Therap 5:1530–1538. doi: 10.1158/1535-7163.MCT-06-0061 CrossRefGoogle Scholar
  12. 12.
    Block G, Patterson B, Subar A (1992) Fruit, vegetables and cancer prevention: a review of the epidemiological evidence. Nutr Cancer 17:1–29Google Scholar
  13. 13.
    Aggarwal BB, Shishodia S (2006) Molecular targets of dietary agents for prevention and therapy of cancer. Biochem Pharmacol 71:1397–1421. doi: 10.1016/j.bcp.2006.02.009 PubMedCrossRefGoogle Scholar
  14. 14.
    Surh YJ, Lee E, Lee JM (1998) Chemoprotective properties of some pungent ingredients present in red pepper and ginger. Mutat Res 402:259–267. doi: 10.1016/S0027-5107(97)00305-9 PubMedGoogle Scholar
  15. 15.
    Buiatti E, Palli D, Decarli A (1989) A case–control study of gastric cancer and diet in Italy. Int J Cancer 444:611–616. doi: 10.1002/ijc.2910440409 CrossRefGoogle Scholar
  16. 16.
    Mozsik G, Szolcsanyi J, Racz I (2005) Gastroprotection induced by capsaicin in healthy subjects. World J Gastroenterol 11:5180–5184PubMedGoogle Scholar
  17. 17.
    Kang JY, Yeoh KG, Chia HP, Lee HP, Chia YW, Guan R (1995) Chilli protective factor against peptic ulcer. Dig Dis Sci 40:576–579. doi: 10.1007/BF02064373 PubMedCrossRefGoogle Scholar
  18. 18.
    Satyanarayana MN (2006) Capsaicin and gastric ulcers. Crit Rev Food Sci Nutr 29:435–473Google Scholar
  19. 19.
    Suzuki T, Iwai K (1994) Constitution of red pepper species: chemistry, biochemistry, pharmacology, and food science of the pungent principle of capsicum species. In: Brosi A (ed) The Alkaloids. Academeic press, New York, pp 227–299Google Scholar
  20. 20.
    Cordell GA, Araujo OE (1993) Capsaicin: identification, nomenclature, and pharmacotherapy. Ann Pharmacother 27:330–336PubMedGoogle Scholar
  21. 21.
    Parrish M (1996) Liquid chromatographic method of determining capsaicinoids in capsicums and their extractives: Collaborative study. J Assoc Anal Chem Intern 79:738–745Google Scholar
  22. 22.
    Busch SH, Burks TF (1983) Hot new pharmacological tool. Trends Pharmacol Sci 4:84–87. doi: 10.1016/0165-6147(83)90297-3 CrossRefGoogle Scholar
  23. 23.
    Taik-koo Y (1999) Update from Asia: Asian studies on cancer chemoprevention. Ann NY Acad Sci 889:157–192. doi: 10.1111/j.1749-6632.1999.tb08734.x CrossRefGoogle Scholar
  24. 24.
    Govindarajan VS, Sathyanarayana MN (1991) Capsicum-production, technology, chemistry, and quality. Part V. Impact on physiology, pharmacology, nutrition and metabolism; structure, pungency, pain and desensitization sequences. Crit Rev Food Sci Nutr 29:435–473PubMedCrossRefGoogle Scholar
  25. 25.
    Monsereenusorn Y (1983) Subchronic toxicity studies of capsaicin and capsicum in rats. Res Commun Chem Pathol Pharmacol 41:95–110PubMedGoogle Scholar
  26. 26.
    Lopez-Carrillo L, Avila HM, Dubrow R (1994) Chili pepper consumption and gastric cancer in Mexico: A case control study. Am J Epidemiol 139:263–271PubMedGoogle Scholar
  27. 27.
    Matucci CM, McCarthy G, Lombardi A, Pignone A, Partsch G (1995) Neurogenic influences in arthritis: potential modification by capsaicin. J Rheumatol 22:1447–1449Google Scholar
  28. 28.
    Sicuteri F, Fusco BM, Marabini S (1989) Beneficial effect of capsaicin application to the nasal mucosa in cluster headache. Clin J Pain 5:49–53. doi: 10.1097/00002508-198903000-00010 PubMedCrossRefGoogle Scholar
  29. 29.
    Watson CP, Evans RJ, Watt VR (1988) Post herpetic neuralgia and topical capsaicin. Pain 33:333–340. doi: 10.1016/0304-3959(88)90292-8 PubMedCrossRefGoogle Scholar
  30. 30.
    Holzer P (1991) Capsaicin: cellular targets, mechanisms of action, and selectivity for thin sensory neurons. Pharmacol Rev 43:143–201PubMedGoogle Scholar
  31. 31.
    Gunthorpe MJ, Szallasi A (2008) Peripheral TRPV1 receptors as targets for drug development: new approaches and mechanisms. Curr Pharm Des 14:32–41. doi: 10.2174/138161208783330754 PubMedCrossRefGoogle Scholar
  32. 32.
    Raisinghani M, Pabbidi RB, Premkumar LS (2005) Activation of transient receptor potential vanilloid 1 (TRPV1) by resiniferatoxin. J Phisiol 567:771–786. doi: 10.1113/jphysiol.2005.087874 CrossRefGoogle Scholar
  33. 33.
    Mori A, Lehmann S, O’Kelly J, Kumagai T, Desmond JC, Pervan M et al (2006) Capsaicin a component of red peppers inhibits the growth of andogen independent, p53 mutant prostate cancer cells. Cancer Res 66:3222–3229. doi: 10.1158/0008-5472.CAN-05-0087 PubMedCrossRefGoogle Scholar
  34. 34.
    Jang JJ, Kim SH, Yun TK (1989) Inhibitory effect of capsaicin on mouse lung development. In Vivo 3:49–53PubMedGoogle Scholar
  35. 35.
    Modly CE, Das M, Don PC, Marcelo CL, Mukhtar H, Bickers DR (1986) Capsaicin as in vitro inhibitor of benzo(a)pyrene metabolism and its DNA binding in human and murine keratinocytes. Drug Metab Dispos 14:413–416PubMedGoogle Scholar
  36. 36.
    Yoshitani SI, Tanaka T, Kohno H, Takashima S (2001) Chemoprevention of azoxymethane-induced rat colon carcinogenesis by dietary capsaicin and rotenone. Int J Oncol 19:929–939PubMedGoogle Scholar
  37. 37.
    Surh YJ, Lee SS (1995) Capsaicin, a double-edged sword: toxicity, metabolism, and chemopreventive potential. Life Sci 56:1845–1855. doi: 10.1016/0024-3205(95)00159-4 PubMedCrossRefGoogle Scholar
  38. 38.
    Teel RW (1991) Effects of capsaicin on rat liver S9-mediated metabolism and DNA binding of aflatoxin. Nutr Cancer 15:27–32PubMedCrossRefGoogle Scholar
  39. 39.
    Sanchez AM, Sanchez MG, Malagarie-Cazenave S, Olea N, Diaz-Laviada I (2006) Induction of apoptosis in prostate tumor PC-3 cells and inhibition of xenograft prostate tumor growth by the vanilloid capsaicin. Apoptosis 11:89–99. doi: 10.1007/s10495-005-3275-z PubMedCrossRefGoogle Scholar
  40. 40.
    Sanchez AM, Malagarie-Cazenave S, Olea N, Vara D, Chiloeches A, Diaz-Laviada I (2007) Apoptosis induced by capsaicin in prostate PC-3 cells involves ceramide accumulation, neutral sphingomyelinase, and JNK activation. Apoptosis 12:2013–2024. doi: 10.1007/s10495-007-0119-z PubMedCrossRefGoogle Scholar
  41. 41.
    Hail N, Lotan R (2002) Examining the role of mitochondrial respiration in vanilloid-induced apoptosis. J Natl Cancer Inst 94:1281–1292PubMedGoogle Scholar
  42. 42.
    Ito K, Nakazato T, Yamato K, Miyakawa Y, Yamada T, Hozumi N et al (2004) Induction of apoptosis in leukemic cells by homovanillic acid derivative capsaicin through oxidative stress implication of phosphorylation of p53 at Ser-15 residue by reactive oxygen species. Cancer Res 64:1071–1078. doi: 10.1158/0008-5472.CAN-03-1670 PubMedCrossRefGoogle Scholar
  43. 43.
    Kang HJ, Soh Y, Kim MS, Lee EJ, Surh YJ, Kim HC et al (2003) Roles of JNK-1 and p38 in selective induction of apoptosis by capsaicin in ras-transformed human breast epithelial cells. Int J Cancer 103:475–482. doi: 10.1002/ijc.10855 PubMedCrossRefGoogle Scholar
  44. 44.
    Lee YS, Kang YS, Lee JS, Nicolova SS, Kim JE (2004) Involvement of NADPH oxidase-mediated generation of reactive oxygen species in the apoptotic cell death by capsaicin in HepG2 human hepatoma cells. Free Radic Res 38:405–412. doi: 10.1080/10715760410001665262 PubMedCrossRefGoogle Scholar
  45. 45.
    Min JK, Han KY, Kim EC, Kim YM, Lee SW, Kim OH et al (2004) Capsaicin inhibits in vitro and in vivo angiogenesis. Cancer Res 64:644–651. doi: 10.1158/0008-5472.CAN-03-3250 PubMedCrossRefGoogle Scholar
  46. 46.
    Surh YJ (2002) More than spice: capsaicin in hot chili pepper makes tumor cells commit suicide. J Natl Cancer Inst 94:1263–1265PubMedGoogle Scholar
  47. 47.
    Srivastava SK, Singh SV (2004) Cell cycle arrest and apoptosis-induced by benzyl isothiocyanate are associated with inhibition of nuclear factor kappa B activation in human pancreatic cancer cells. Carcinogenesis 25:1701–1709. doi: 10.1093/carcin/bgh179 PubMedCrossRefGoogle Scholar
  48. 48.
    Zhang R, Loganathan S, Humphreys I, Srivastava SK (2006) Benzyl isothiocyanate induced DNA damage causes G2/M cell cycle arrest and apoptosis in human pancreatic cancer cells. J Nutr 136:2728–2734PubMedGoogle Scholar
  49. 49.
    Shi Y, Sahu RP, Srivastava SK (2008) Triphala inhibits both in vitro and in vivo xenograft growth of pancreatic tumor cells by inducing apoptosis. BMC Cancer 8:294. doi: 10.1186/1471-2407-8-294 PubMedCrossRefGoogle Scholar
  50. 50.
    Srivastava SK, Xiao D, Lew KL, Hershberger P, Kokkinakis DM, Johnson CS et al (2003) Allyl isothiocyanate, a natural constituent of cruciferous vegetables inhibits growth of PC-3 human prostate cancer xenografts in vivo in association with down-regulation of Bcl-2. Carcinogenesis 24:665–670. doi: 10.1093/carcin/bgg123 CrossRefGoogle Scholar
  51. 51.
    Singh SV, Srivastava SK, Choi S, Lew KL, Antosiewitz J, Xiao D et al (2005) Sulphoraphane-induced cell death in human prostate cancer cells is initiated by reative oxygen species. J Biol Chem 280:19911–19924. doi: 10.1074/jbc.M412443200 PubMedCrossRefGoogle Scholar
  52. 52.
    Shen H, Liu Z (2006) JNK signaling pathway is a key modulator in cell death mediated by reactive oxygen and nitrogen species. Free Radic Biol Med 40:928–939. doi: 10.1016/j.freeradbiomed.2005.10.056 PubMedCrossRefGoogle Scholar
  53. 53.
    Council of Europe (2001) Committee of experts on flavoring substances. Datasheet on capsaicin, 612Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Ruifen Zhang
    • 1
    • 2
  • Ian Humphreys
    • 2
  • Ravi P. Sahu
    • 3
  • Yan Shi
    • 3
  • Sanjay K. Srivastava
    • 1
    • 2
    • 3
    Email author
  1. 1.Department of PharmacologyUniversity of Pittsburgh School of MedicinePittsburghUSA
  2. 2.University of Pittsburgh Cancer InstituteUniversity of Pittsburgh School of MedicinePittsburghUSA
  3. 3.Department of Pharmaceutical Sciences and Cancer Biology Center, School of PharmacyTexas Tech University Health Sciences CenterAmarilloUSA

Personalised recommendations