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Curcumin sensitizes non-small cell lung cancer cell anoikis through reactive oxygen species-mediated Bcl-2 downregulation

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Abstract

Anoikis, an apoptosis triggered by loss of cell anchorage, has been shown to be a principal mechanism of inhibition of tumor metastasis. Recently, anti-apoptotic Bcl-2 and Cav-1 proteins have been demonstrated to be highly associated with tumor metastasis and apoptosis resistance. Curcumin, a major active component of turmeric, Curcuma longa, has been shown to inhibit neoplastic evolution and tumor progression; however, the underlying mechanisms are unclear. In this study, we investigated the effect of curcumin on cell anoikis as a possible mechanism of anti-tumorigenic action of curcumin, and evaluated the potential role of Bcl-2 and Cav-1 in this process. Our results showed that ectopic expression of either Bcl-2 or Cav-1 induced anoikis resistance of lung carcinoma H460 cells. Curcumin downregulated Bcl-2 protein during anoikis and sensitized the cells to detachment-induced apoptosis, whereas it had no significant effect on Cav-1 protein expression. Bcl-2 down-regulation as well as anoikis enhancement by curcumin were inhibited by superoxide anion scavenger, Mn(III)tetrakis(4-benzoic acid) porphyrin chloride, but were unaffected by other ROS scavengers including catalase and deferoxamine, suggesting that superoxide anion is a key player in the downregulation of Bcl-2 by curcumin. Furthermore, we provided evidence that curcumin decreased Bcl-2 level through ubiquitin-proteasomal degradation which sensitized cells to detachment-induced apoptosis. These findings indicate a novel pathway for curcumin regulation of Bcl-2 and provide a key mechanism of anoikis regulation that may be exploited for metastatic cancer treatment.

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Abbreviations

XTT:

2,3-Bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5 carboxanilide inner salt

DHE:

Dihydroethidium

H2DCF-DA:

Dihydrodichlorofluorescein diacetate

MnTBAP:

Mn(III)tetrakis(4-benzoic acid) porphyrin chloride

Def:

Deferoxamine

Cat:

Catalase

polyHEMA:

Poly(2-hydroxy ethylmethacrylate)

Cav-1:

Caveolin-1

References

  1. Han SS, Chung ST, Robertson DA, Ranjan D, Bondada S (1999) Curcumin causes the growth arrest and apoptosis of B cell lymphoma by downregulation of egr-1, c-myc, bcl-XL, NF-kappa B, and p53. Clin Immunol 93:152–161. doi:10.1006/clim.1999.4769

    Article  PubMed  CAS  Google Scholar 

  2. Simon A, Allais DP, Duroux JL, Basly JP, Durand-Fontanier S, Delage C (1998) Inhibitory effect of curcuminoids on MCF-7 cell proliferation and structure-activity relationships. Cancer Lett 129:111–116. doi:10.1016/S0304-3835(98)00092-5

    Article  PubMed  CAS  Google Scholar 

  3. Bush JA, Cheung KJJ, Li G (2001) Curcumin induces apoptosis in human melanoma cells through a Fas receptor/caspase-8 pathway independent of p53. Exp Cell Res 271:305–314. doi:10.1006/excr.2001.5381

    Article  PubMed  CAS  Google Scholar 

  4. Bachmeier B, Nerlich AG, Iancu CM, Cilli M, Schleicher E, Vené R, Dell’Eva R, Jochum M, Albini A, Pfeffer U (2007) The chemopreventive polyphenol curcumin prevents hematogenous breast cancer metastases in immunodeficient mice. Cell Physiol Biochem 19:137–152

    Article  PubMed  CAS  Google Scholar 

  5. Chen HW, Lee JY, Huang JY, Wang CC, Chen WJ, Su SF, Huang CW, Ho CC, Chen JJ, Tsai MF, Yu SL, Yang PC (2008) Curcumin inhibits lung cancer cell invasion and metastasis through the tumor suppressor HLJ1. Cancer Res 68:7428–7438. doi:10.1158/0008-5472.CAN-07-6734

    Article  PubMed  CAS  Google Scholar 

  6. Yoshino M, Haneda M, Naruse M, Htay HH, Tsubouchi R, Qiao SL, Li WH, Murakami K, Yokochi T (2004) Prooxidant activity of curcumin: copper-dependent formation of 8-hydroxy-2′-deoxyguanosine in DNA and induction of apoptotic cell death. Toxicol In Vitro 18:783–789. doi:10.1016/j.tiv.2004.03.009

    Article  PubMed  CAS  Google Scholar 

  7. Cao J, Jia L, Zhou HM, Liu Y, Zhong LF (2006) Mitochondrial and nuclear DNA damage induced by curcumin in human hepatoma G2 cells. Toxicol Sci 91:476–483. doi:10.1093/toxsci/kfj153

    Article  PubMed  CAS  Google Scholar 

  8. Chanvorachote P, Pongrakhananon V, Wannachaiyasit S, Luanpitpong S, Rojanasakul Y, Nimmanit U (2009) Curcumin sensitizes lung cancer cells to cisplatin-induced apoptosis through superoxide anion-mediated Bcl-2 degradation. Cancer Invest 26:624–635. doi:10.1080/07357900802653472

    Article  CAS  Google Scholar 

  9. Grossmann J (2002) Molecular mechanisms of “detachment-induced apoptosis-anoikis”. Apoptosis 7:247–260. doi:10.1023/A:1015312119693

    Article  PubMed  CAS  Google Scholar 

  10. Mehlen P, Puisieux A (2006) Metastasis: a question of life or death. Nat Rev Cancer 6:449–458. doi:10.1038/nrc1886

    Article  PubMed  CAS  Google Scholar 

  11. Hanahan D, Weiberg RA (2000) The hallmarks of cancer. Cell 100:57–70. doi:10.1016/S0092-8674(00)81683-9

    Article  PubMed  CAS  Google Scholar 

  12. Frisch SM, Screaton RA (2001) Anoikis Mechanisms. Curr Opin Cell Biol 13:555–562. doi:10.1016/S0955-0674(00)00251-9

    Article  PubMed  CAS  Google Scholar 

  13. Chen Y, Kuo C, Tsai Y, Yu C, Wang G, Liao H (2008) Norcantharidin induces anoikis through Jun-N-terminal kinase activation in CT26 colorectal cancer cells. Anticancer Drugs 19:55–64. doi:10.1097/CAD.0b013e3282f18826

    Article  PubMed  CAS  Google Scholar 

  14. Dufour G, Demers MJ, Gagne D, Dydensborg AB, Teller IC, Bouchard V, Degongre I, Beaulieu JF, Cheng JQ, Fujita N, Tsuruo T, Vallee K, Vachon PH (2004) Human intestinal epithelial cell survival and anoikis: differentiation state distinct regulation and roles of protein kinase B/Akt isoforms. J Biol Chem 279:44113–44122. doi:10.1074/jbc.M405323200

    Article  PubMed  CAS  Google Scholar 

  15. Almeida EA, Ilic D, Han Q, Hauck CR, Jin F, Kawakatsu H, Schlaepfer DD, Damsky CH (2000) Matrix survival signaling: from fibronectin via focal adhesion kinase to c-Jun NH(2)-terminal kinase. J Cell Biol 149:741–754

    Article  PubMed  CAS  Google Scholar 

  16. Collins NL, Reginato MJ, Paulus JK, Sgroi DC, Joshua L, Brugge JS (2005) G1/S cell cycle arrest provides anoikis resistance through Erk-mediated Bim suppression. Mol Cell Biol 25:5282–5291. doi:10.1128/MCB.25.12.5282-5291.2005

    Article  PubMed  CAS  Google Scholar 

  17. Galante JM, Mortenson MM, Bowles TL, Virudachalam S, Bold RJ (2009) Erk/Bcl-2 pathway in the resistance of pancreatic cancer to anoikis. J Surg Res 152:18–25. doi:10.1016/j.jss.2008.05.017

    Article  PubMed  CAS  Google Scholar 

  18. Martin SS, Vuori K (2004) Regulation of Bcl-2 proteins during anoikis and amorphosis. Biochim Biophys Acta 1692:145–157. doi:10.1016/j.bbamcr.2004.02.008

    PubMed  CAS  Google Scholar 

  19. Reginato MJ, Mills KR, Paulus JK, Lynch DK, Sgroi DC, Debnath J, Muthuswamy SK, Brugg JS (2003) Intergrins and EGFR coordinately regulate the pro-apoptotic protein Bim to prevent anoikis. Nat Cell Biol 35:733–740. doi:10.1038/ncb1026

    Article  CAS  Google Scholar 

  20. Schmelzle T, Mailluex AA, Overholtzer M, Carroll JS, Solimini NL, Lightcap ES, Veiby OP, Brugge JS (2006) Functional role and oncogene-regulated expression of the BH3-only factor Bmf in mammary epithelial anoikis and morphogenesis. Proc Natl Acad Sci USA 104:3787–3792. doi:10.1073/pnas.0700115104

    Article  CAS  Google Scholar 

  21. Valentijn AJ, Metcalfe AD, Kott J, Streuli CH, Gilmore AP (2003) Spatial and temporal change in Bax subcellular localization during anoikis. J Cell Biol 162:599–612. doi:10.1083/jcb.200302154

    Article  PubMed  CAS  Google Scholar 

  22. Valentijn AJ, Gilmore AP (2004) Translocation of full-length Bid to mitochondria during anoikis. J Biol Chem 279:32848–32857. doi:10.1074/jbc.M313375200

    Article  PubMed  CAS  Google Scholar 

  23. Frankel A, Rosen K, Filmus J, Kerbel RS (2001) Induction of anoikis and suppression of human ovarian tumor growth in vivo by down-regalation of Bcl-XL. Cancer Res 61:4837–4841

    PubMed  CAS  Google Scholar 

  24. Scorrano L, Korsmeyer SJ (2003) Mechanisms of cytochrome c release by proapoptotic BCL-2 family members. Biochem Biophys Res Commun 304:437–444

    Article  PubMed  CAS  Google Scholar 

  25. Grossmann J, Walther K, Artinger M, Kiessling S, Scholmerich J (2001) Apoptotic signaling during initiation of detachment-induced apoptosis (‘Anoikis’) of primary human intestinal epithelial cells. Cell Growth Diff 12:147–155

    PubMed  CAS  Google Scholar 

  26. Lin D, Feng J, Chen W (2008) Bcl-2 and caspase-8 related anoikis resistance in human osteosarcoma MG-63 cells. Cell Biol Int 32:1199–1206. doi:10.1016/j.cellbi.2008.07.002

    Article  PubMed  CAS  Google Scholar 

  27. Dimmeler S, Breitschopf K, Haendeler J, Zeiher AM (1999) Dephosphorylation targets Bcl-2 for ubiquitin-dependent degradation: a link between the apoptosome and the proteasome pathway. J Exp Med 189:1815–1822

    Article  PubMed  CAS  Google Scholar 

  28. Chanvorachote P, Nimmanit U, Stehlik C, Wang L, Jiang B, Ongpipatanakul B, Rojanasakul Y (2006) Nitric oxide regulates cell sensitivity to cisplatin-induced apoptosis through S-nitrosylation and inhibition of Bcl-2 ubiquitination. Cancer Res 66:6353–6360. doi:10.1158/0008-5472.CAN-05-4533

    Article  PubMed  CAS  Google Scholar 

  29. Haendeler J, Messmer UK, Brune B, Neugebauer E, Dimmeler S (1996) Endotoxic shock leads to apoptosis in vivo and reduces Bcl-2. Shock 6:405–409

    Article  PubMed  CAS  Google Scholar 

  30. Paradis E, Doulliard H, Koutroumanis M, Goodyer C, Leblanc A (1996) Amyloid beta peptide of alzheimer’s disease downregulates Bcl-2 and upregulates Bax expression in human neurons. J Neurosci 16:7533–7539

    PubMed  CAS  Google Scholar 

  31. Yanamadala V, Negoro H, Gunaratnam L, Kong T, Denker BM (2007) Galpha12 stimulates apoptosis in epithelial cells through JNK1-mediated Bcl-2 degradation and up-regulation of IkappaBalpha. J Biol Chem 282:24352–24363. doi:10.1074/jbc.M702804200

    Article  PubMed  CAS  Google Scholar 

  32. Williams TM, Lisanti MP (2005) Caveolin-1 in oncogenic transformation, cancer, and metastasis. Am J Physiol Cell Physiol 288:C494–C506. doi:10.1152/ajpcell.00458.2004

    Article  PubMed  CAS  Google Scholar 

  33. Yang G, Truong LD, Timme TL, Ren C, Wheeler TM, Park SH, Nasu Y, Bangma CH, Kattan MW, Scardino PT, Thompson TC (1998) Elevated expression of caveolin is associated with prostate and breast cancer. Clin Cancer Res 4:1873–1880

    PubMed  CAS  Google Scholar 

  34. Ravid D, Maor S, Werner H, Liscovitch M (2006) Caveolin-1 inhibits anoikis and promotes survival signaling in cancer cells. Adv Enzyme Regul 46:163–175. doi:10.1016/j.advenzreg.2006.01.022

    Article  PubMed  CAS  Google Scholar 

  35. Fiucci G, Ravid D, Reich R, Liscovitch M (2002) Caveolin-1 inhibits anchorage-independent growth, anoikis and invasiveness in MCF-7 human breast cancer cells. Oncogene 21:2365–2375. doi:10.1038/sj/onc/1205300

    Article  PubMed  CAS  Google Scholar 

  36. Ravid D, Maor S, Werner H, Liscovitch M (2005) Caveolin-1 inhibits cell detachment-induced p53 activation and anoikis by upregulation of insulin-like growth factor-I receptors and signaling. Oncogene 24:1338–1347. doi:10.1038/sj.onc.1208337

    Article  PubMed  CAS  Google Scholar 

  37. Sandler AB, Nemunaitis J, Denham C, vonPawel J, Cormier Y, Gatzemeier U, Mattson K, Ch Manegold, Palmer MC, Gregor A, Nguyen B, Niyikiza C, Einhorn LH (2000) Phase III trial of gemcitabine plus cisplatin versus cisplatin alone in patients with locally advanced or metastatic non-small-cell lung cancer. J Clin Oncol 18:122–130

    PubMed  CAS  Google Scholar 

  38. Gan PP, Pasquier E, Kavallaris M (2007) Class III B-tubulin mediates sensitivity to chemotherapeutic drugs in non-small cell lung cancer. Cancer Res 67:9356–9363. doi:10.1158/0008-5472.CAN-07-0509

    Article  PubMed  CAS  Google Scholar 

  39. Bhaumik S, Anjum R, Rangaraj N, Pardhasaradhi BVV, Khar A (1999) Curcumin mediated apoptosis in AK-5 tumor cells involves the production of reactive oxygen intermediates. FEBS Lett 456:311–314. doi:10.1016/S0014-5793(99)00969-2

    Article  PubMed  CAS  Google Scholar 

  40. Syng-ai C, Kumari A, Khar A (2004) Effect of curcumin on normal and tumor cells: role of glutathione and bcl-2. Mol Cancer Ther 3:1101–1108

    PubMed  CAS  Google Scholar 

  41. Breitshopf K, Haendeler J, Malchow P, Zeiher AM, Dimmeler S (2000) Posttranslational modification of bcl-2 facilitates its proteasome-dependent degradation: molecular characterization of the involved signaling pathway. Mol Cell Biol 20:1886–1896

    Article  Google Scholar 

  42. Pillai GR, Srivastava AS, Hassanein TI, Chauhan DP, Carrier E (2004) Induction of apoptosis in human lung cancer cells by curcumin. Cancer Lett 208:163–170. doi:10.1016/j.canlet.2004.01.008

    Article  CAS  Google Scholar 

  43. Notarbartolo M, Poma P, Perri D, Dusonchet L, Cervello M, D’Alessandro N (2005) Antitumor effects of curcumin, alone or in combination with cisplatin or doxorubicin, on human hepatic cancer cells. Analysis of their possible relationship to changes in NF-kB activation levels and in IAP gene expression. Cancer Lett 244:53–65

    Google Scholar 

  44. Weir NM, Selvendiran K, Kutala VK, Tong L, Vishwanath S, Rajaram M, Tridandapani S, Anant S, Kuppusamy P (2007) Curcumin induced G2/M arrest and apoptosis in cisplatin-resistant human ovarian cancer cells by modulating Akt and p38 MAPK. Cancer Biol Ther 6:178–184

    Article  PubMed  CAS  Google Scholar 

  45. Du B, Jiang L, Xia Q, Zhong L (2006) Synergistic inhibitory effect of curcumin and 5-fluorouracil on the growth of the human colon cancer cell line HT-29. Chemotherapy 52:23–28. doi:10.1159/000090238

    Article  PubMed  CAS  Google Scholar 

  46. Deeb D, Jiang H, Gao X, Hafner MS, Wong H, Divine G, Chapman RA, Dulchavsky SA, Gautam SC (2004) Curcumin sensitizes prostate cancer cells to tumor necrosis factor-related apoptosis-inducing ligand/Apo2L by inhibiting nuclear factor-kappaB through suppression of IkappaB alpha phosphorelation. Mol Cancer Ther 3:803–812

    PubMed  CAS  Google Scholar 

  47. Jung EM, Lim JH, Lee TJ, Park JW, Choi KS, Kwon TK (2005) Curcumin sensitizes tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis through reactive oxygen species-mediated upregulation of death receptor 5 (DR5). Carcinogenesis 26:1905–1913. doi:10.1093/carcin/bgi167

    Article  PubMed  CAS  Google Scholar 

  48. Sloan EK, Stanley KL, Anderson RL (2004) Caveolin-1 inhibits breast cancer growth and metastasis. Oncogene 23:7893–7897. doi:10.1038/sj.onc.1208062

    Article  PubMed  CAS  Google Scholar 

  49. Cantiani L, Manara MC, Zucchini C, Sanctis PD, Zuntini M, Valvassori L, Serra M, Olivero M, Renzo MFD, Colombo MP, Picci P, Scotlandi K (2007) Caveolin-1 reduces osteosarcoma metastases by inhibiting c-Src activity and Met signaling. Cancer Res 67:7674–7685. doi:10.1158/0008-5472.CAN-06-4697

    Article  Google Scholar 

  50. Gu D, Li H, Wang Z, Chen Q, Jiang J, Zhu H (2009) Caveolin-1 inhibits the growth of human laryngeal squamous cell carcinoma and down regulates EGFR-MAPKs signaling pathway. Laryngoscope 117:1782–1789. doi:10.1097/MLG.0b013e31811edd31

    Article  CAS  Google Scholar 

  51. Thayyullathil F, Chathoth S, Hago A, Patel M, Galadari S (2008) Rapid reactive oxygen species (ROS) generation induced by curcumin leads to caspase-dependent and -independent apoptosis in L929 cells. Free Radic Biol Med 45:1403–1412. doi:10.1016/j.freeradbiomed.2008.08.014

    Article  PubMed  CAS  Google Scholar 

  52. Khar A, Ali AM, Pardhasaradhi BVV, Varalakshmi C, Anjum R, Kumari AL (2001) Induction of stress response renders human tumor cell lines resistant to curcumin-mediated apoptosis: role of reactive oxygen intermediates. Cell Stress Chaperones 6:368–376

    Article  PubMed  CAS  Google Scholar 

  53. Wang L, Chanvorachote P, Toledo D, Stehlik C, Mercer RR, Castranova V, Rojanasakul Y (2008) Peroxide is a key mediator of Bcl-2 down-regulation and apoptosis induction by cisplatin in human lung cancer cells. Mol Pharmacol 73:119–127. doi:10.1124/mol.107.040873

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by the Thailand Research Fund (MG5080134; Pithi Chanvorachote and RGJ 5.Q.CU/50/A.1).

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Authors and Affiliations

  1. Pharmaceutical Technology (International) Program, Chulalongkorn University, Bangkok, 10330, Thailand

    Varisa Pongrakhananon & Sudjit Luanpitpong

  2. National Nanotechnology Center, Pathumthani, 12120, Thailand

    Ubonthip Nimmannit

  3. Department of Pharmaceutical Sciences, West Virginia University, Morgantown, WV, 26506, USA

    Yon Rojanasakul

  4. Department of Pharmacology and Physiology, Chulalongkorn University, Bangkok, 10330, Thailand

    Pithi Chanvorachote

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  1. Varisa Pongrakhananon
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  2. Ubonthip Nimmannit
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  3. Sudjit Luanpitpong
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  4. Yon Rojanasakul
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  5. Pithi Chanvorachote
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Correspondence to Yon Rojanasakul or Pithi Chanvorachote.

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Pongrakhananon, V., Nimmannit, U., Luanpitpong, S. et al. Curcumin sensitizes non-small cell lung cancer cell anoikis through reactive oxygen species-mediated Bcl-2 downregulation. Apoptosis 15, 574–585 (2010). https://doi.org/10.1007/s10495-010-0461-4

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