Skip to main content
SpringerLink
Log in

Resveratrol induces mitochondria-mediated AIF and to a lesser extent caspase-9-dependent apoptosis in human lung adenocarcinoma ASTC-a-1 cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Resveratrol (RV), a natural plant polyphenol widely present in foods such as grapes, wine, and peanuts, has an ability to inhibit various stages of carcinogenesis in vitro and in vivo. In this report, we explored the roles of intrinsic and extrinsic apoptotic pathways during RV-induced apoptosis in human lung adenocarcinoma (ASTC-a-1) cells. After exposure of cells to different concentrations of RV, we found that RV induced concentration-dependent apoptosis. Fluorometric substrates assay and western blotting (WB) analysis showed that caspase-8 was not activated, which was further verified by monitoring the cleavage of Bid to tBid using fluorescence resonance energy transfer (FRET) microscopy imaging inside single living cells, indicating that extrinsic apoptotic pathway was not involved in RV-induced apoptosis. In addition, inhibition of caspases-3 or -9 but not caspase-8 using the specific inhibitors of caspases modestly but significantly attenuated RV-induced apoptosis. Moreover, flow cytometry (FCM) analysis showed that RV treatment induced time-dependent loss of mitochondrial membrane potential (∆ψm), in combination with the activation of caspases-3 and -9; we therefore concluded that RV-induced apoptosis involved the intrinsic apoptotic pathway. It is noteworthy that RV treatment induced translocation of AIF from mitochondria to nucleus in a time dependent manner, and that knockdown of AIF remarkably attenuated RV-induced apoptosis. Collectively, our findings demonstrate that RV induces caspase-8-independent apoptosis via AIF and to a lesser extent caspase-9-dependent mitochondrial pathway in ASTC-a-1 cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

AIF:

Apoptosis-inducing factor

CCK-8:

Cell Counting Kit

Endo G:

Endonuclease G

FCM:

Flow cytometry

FRET:

Fluorescence resonance energy transfer

HtrA2/Omi:

High temperature requirement protein A2

Rho123:

Rhodamine123

RV:

Resveratrol

Smac/Diablo:

Second mitochondrial activator of caspases

STS:

Staurosporine

ΔΨm :

Mitochondrial membrane potential

ECFP:

Enhanced cyan fluorescent protein

Venus:

A mutant of yellow fluorescent protein (YFP)

References

  1. Fennell DA (2005) Caspase regulation in non-small cell lung cancer and its potential for therapeutic exploitation. Clin Cancer Res 11:2097–2105

    Article  CAS  PubMed  Google Scholar 

  2. Ghavami S, Hashemi M, Ande SR, Yeganeh B, Xiao W, Eshraghi M, Bus CJ, Kadkhoda K, Wiechec E, Halayko AJ, Los M (2009) Apoptosis and cancer: mutations within caspase genes. J Med Genet 46:497–510

    Article  CAS  PubMed  Google Scholar 

  3. Ravagnan L, Roumier T, Kroemer G (2002) Mitochondria—the killer organelles and their weapons. J Cell Physiol 192:131–137

    Article  CAS  PubMed  Google Scholar 

  4. Sareen D, van Ginkel PR, Takach JC, Darjatmoko SR, Mohiuddin A, Albert DM, Polans AS (2006) Mitochondria as the primary target of resveratrol-induced apoptosis in human retinoblastoma cells. Invest Ophthalmol Vis Sci 47:3708–3716

    Article  PubMed  Google Scholar 

  5. Lee TJ, Kim EJ, Kim S, Jung EM, Park JW, Jeong SH, Park SE, Yoo YH, Kwon TK (2006) Caspase-dependent and caspase-independent apoptosis induced by evodiamine in human leukemic U937 cells. Mol Cancer Ther 5:2398–2407

    Article  CAS  PubMed  Google Scholar 

  6. Debatin KM (2004) Apoptosis pathways in cancer and cancer therapy. Cancer Immunol Immun 53:153–159

    Google Scholar 

  7. Mohan J, Gandhi AA, Bhavya BC, Rashmi R, Karunagaran D (2006) Caspase-2 triggers bax-bak-dependent and -independent cell death in colon cancer cells treated with resveratrol. J Biol Chem 281:17599–17611

    Article  CAS  PubMed  Google Scholar 

  8. Athar M, Back JH, Kopelovich L, Bickers DR, Kim AL (2009) Multiple molecular targets of resveratrol: anti-carcinogenic mechanisms. Arch Biochem Biophys 486:95–102

    Article  CAS  PubMed  Google Scholar 

  9. Shankar S, Siddiqui I, Srivastava RK (2007) Molecular mechanisms of resveratrol (3,4,5-trihydroxy-transstilbene) and its interaction with TNF-related apoptosis inducing ligand (TRAIL) in androgen-insensitive prostate cancer cells. Mol Cell Biochem 304:273–285

    Article  CAS  PubMed  Google Scholar 

  10. Souza IC, Martins LA, Coelho BP, Grivicich I, Guaragna RM, Gottfried C, Borojevic R, Guma FC (2008) Resveratrol inhibits cell growth by inducing cell cycle arrest in activated hepatic stellate cells. Mol Cell Biochem 315:1–7

    Article  CAS  PubMed  Google Scholar 

  11. Bishayee A (2009) Cancer prevention and treatment with resveratrol: from rodent studies to clinical trials. Cancer Prev Res 2:409–418

    Article  CAS  Google Scholar 

  12. Joe AK, Liu H, Suzui M, Vural ME, Xiao D, Weinstein IB (2002) Resveratrol induces growth inhibition, S-phase arrest, apoptosis, and changes in biomarker expression in several human cancer cell lines. Clin Cancer Res 8:893–903

    CAS  PubMed  Google Scholar 

  13. Srivastava RK, Unterman TG, Shankar S (2010) FOXO transcription factors and VEGF neutralizing antibody enhance antiangiogenic effects of resveratrol. Mol Cell Biochem 337:201–212

    Article  CAS  PubMed  Google Scholar 

  14. Byun HS, Song JK, Kim YR, Piao L, Won M, Park KA, Choi BL, Lee H, Hong JH, Park J, Seok JH, Lee YJ, Kang SW, Hur GM (2008) Caspase-8 has an essential role in resveratrol-induced apoptosis of rheumatoid fibroblast-like synoviocytes. Rheumatology 47:301–308

    Article  CAS  PubMed  Google Scholar 

  15. Dörrie J, Gerauer H, Wachter Y, Zunino SJ (2001) Resveratrol induces extensive apoptosis by depolarizing mitochondrial membranes and activating caspase-9 in acute lymphoblastic leukemia cells. Cancer Res 61:4731–4739

    PubMed  Google Scholar 

  16. Kim MY, Trudel LJ, Wogan GN (2009) Apoptosis induced by capsaicin and resveratrol in colon carcinoma cells requires nitric oxide production and caspase activation. Anticancer Res 29:3733–3740

    CAS  PubMed  Google Scholar 

  17. Pozo-Guisado E, Merino JM, Mulero-Navarro S, Lorenzo-Benayas MJ, Centeno F, Alvarez-Barrientos A, Fernandez-Salguero PM (2005) Resveratrol-induced apoptosis in MCF-7 human breast cancer cells involves a caspase-independent mechanism with down-regulation of Bcl-2 and NF-kappaB. Int J Cancer 115:74–84

    Article  CAS  PubMed  Google Scholar 

  18. Whyte L, Huang YY, Torres K, Mehta RG (2007) Molecular mechanisms of resveratrol action in lung cancer cells using dual protein and microarray analyses. Cancer Res 67:12007–12017

    Article  CAS  PubMed  Google Scholar 

  19. Kim YA, Lee WH, Choi TH, Rhee SH, Park KY, Choi YH (2003) Involvement of p21WAF1/CIP1, pRB, Bax and NF-κB in induction of growth arrest and apoptosis by resveratrol in human lung carcinoma A549 cells. Int J Oncol 23:1143–1149

    CAS  PubMed  Google Scholar 

  20. Takemoto K, Nagai T, Miyawaki A, Miura M (2003) Spatio-temporal activation of caspase revealed by indicator that is insensitive to environmental effects. J Cell Biol 160:235–243

    Article  CAS  PubMed  Google Scholar 

  21. Onuki R, Nagasaki A, Kawasaki H, Baba T, Uyeda TQP, Taira K (2002) Confirmation by FRET in individual living cells of the absence of significant amyloid-mediated caspase 8 activation. Proc Natl Acad Sci USA 99:14716–14721

    Article  CAS  PubMed  Google Scholar 

  22. Tsuruta F, Masuyama N, Gotoh Y (2002) The phosphatidylinositol 3-kinase (PI3K)-Akt pathway suppresses Bax translocation to mitochondria. J Biol Chem 277:14040–14047

    Article  CAS  PubMed  Google Scholar 

  23. Wu Y, Xing D, Chen WR (2006) Single cell FRET imaging for determination of pathway of tumor cell apoptosis induced by photofrin-PDT. Cell Cycle 5:729–734

    Article  CAS  PubMed  Google Scholar 

  24. Lu Y, Chen T, Wang XP, Qu JL, Chen M (2010) The JNK inhibitor SP600125 enhances dihydroartemisinin-induced apoptosis by accelerating Bax translocation into mitochondria in human lung adenocarcinoma cells. FEBS Lett 584:4019–4026

    Article  CAS  PubMed  Google Scholar 

  25. Wang LX, Chen TS, Qu JL (2009) Photobleaching-based quantitative analysis of fluorescence resonance energy transfer inside single living cell. J Fluoresc 20:27–35

    Article  PubMed  Google Scholar 

  26. Zhang YJ, Xing D, Liu L (2009) PUMA promotes bax translocation by both directly activating bax and antagonizing bcl-xl during UV-induced apoptosis. Mol Biol Cell 20:3077–3087

    Article  CAS  PubMed  Google Scholar 

  27. Zinkel SS, Hurov KE, Ong C, Abtahi FM, Gross A, Korsmeyer SJ (2005) A role for proapoptotic BID in the DNA-damage response. Cell 122:579–591

    Article  CAS  PubMed  Google Scholar 

  28. Zha J, Weiler S, Oh KJ, Wei MC, Korsmeyer S (2000) Posttranslational N-myristoylation of BID as a molecular switch for targeting mitochondria and apoptosis. Science 290:1761–1765

    Article  CAS  PubMed  Google Scholar 

  29. Li YL, Xing D, Chen Q, Chen WR (2010) Enhancement of chemotherapeutic agent-induced apoptosis by inhibition of NF-jB using ursolic acid. Int J Cancer 127:462–473

    Article  CAS  PubMed  Google Scholar 

  30. Carney DN (2002) Lung cancer-time to move on from chemotherapy. New Engl J Med 346:126–128

    Article  PubMed  Google Scholar 

  31. Ganapathy S, Chen Q, Singh KP, Shankar S, Srivastava RK (2010) Resveratrol enhances antitumor activity of TRAIL in prostate cancer xenografts through activation of FOXO transcription factor. PLoS One 5:e15627

    Article  CAS  PubMed  Google Scholar 

  32. Chen Q, Ganapathy S, Singh KP, Shankar S, Srivastava RK (2010) Resveratrol induces growth arrest and apoptosis through activation of FOXO transcription factors in prostate cancer cells. PLoS One 5:e15288

    Article  CAS  PubMed  Google Scholar 

  33. Frisch SM (2008) Caspase-8: fly or die. Cancer Res 68:4491–4493

    Article  CAS  PubMed  Google Scholar 

  34. Lovell JF, Billen LP, Bindner S, Shamas-Din A, Fradin C, Leber B, Andrews DW (2008) Membrane binding by tbid initiates an ordered series of events culminating in membrane permeabilization by bax. Cell 135:1074–1084

    Article  CAS  PubMed  Google Scholar 

  35. Lockshin RA, Zakeri Z (2004) Apoptosis, autophagy, and more. Int J Biochem Cell B 36:2405–2419

    Article  CAS  Google Scholar 

  36. Guha P, Dey A, Dhyani MV, Sen R, Chatterjee M, Chattopadhyay S, Bandyopadhyay SK (2010) Calpain and caspase orchestrated death signal to accomplish apoptosis induced by resveratrol and its novel analog hydroxystilbene-1 [correction of hydroxstilbene-1] in cancer cells. J Pharmacol Exp Ther 334:381–394

    Article  CAS  PubMed  Google Scholar 

  37. Van Ginkel PR, Darjatmoko SR, Sareen D, Subramanian L, Bhattacharya S, Lindstrom MJ, Albert DM, Polans AS (2008) Resveratrol inhibits uveal melanoma tumor growth via early mitochondrial dysfunction. Invest Ophthalmol Vis Sci 49:1299–1306

    Article  PubMed  Google Scholar 

  38. Green D, Kroemer G (1998) The central executioners of apoptosis: caspases or mitochondria? Trends Cell Biol 8:267–271

    Article  CAS  PubMed  Google Scholar 

  39. Ma XD, Tian XM, Huang XX, Yan F, Qiao DF (2007) Resveratrol-induced mitochondrial dysfunction and apoptosis are associated with Ca2+ and mCICR-mediated MPT activation in HepG2 cells. Mol Cell Biochem 302:99–109

    Article  CAS  PubMed  Google Scholar 

  40. Sareen D, Darjatmoko SR, Albert DM, Polans AS (2007) Mitochondria, calcium, and calpain are key mediators of resveratrol-induced apoptosis in breast cancer. Mol Pharmacol 72:1466–1475

    Article  CAS  PubMed  Google Scholar 

  41. Mahyar-Roemer M, Kohler H, Roemer K (2002) Role of Bax in resveratrol induced apoptosis of colorectal carcinoma cells. BMC Cancer 2:27

    Article  PubMed  Google Scholar 

  42. Jiang H, Zhang L, Kuo J, Kuo K, Gautam SC, Groc L, Rodriguez AI, Koubi D, Hunter TJ, Corcoran GB, Seidman MD, Levine RA (2005) Resveratrol-induced apoptotic death in human U251glioma cells. Mol Cancer Ther 4:554–561

    Article  CAS  PubMed  Google Scholar 

  43. Kroemer G, Galluzzi L, Brenner C (2007) Mitochondrial membrane permeabilization in cell death. Physiol Rev 87:99–163

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by the National Natural Science Foundation of China (Grant Nos. 31071218 and 81071491).

Author information

Authors and Affiliations

  1. MOE Key Libratory of Laser Life Science, Institute of Laser Life Science, South China Normal University, Guangzhou, 510631, China

    Weiwei Zhang

  2. Department of Anesthesiology, First Clinical Medical College, Jinan University, Guangzhou, 510632, China

    Xiaoping Wang

  3. Institute of Laser Life Science, South China Normal University, Guangzhou, 510631, China

    Tongsheng Chen

Authors
  1. Weiwei Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoping Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tongsheng Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tongsheng Chen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, W., Wang, X. & Chen, T. Resveratrol induces mitochondria-mediated AIF and to a lesser extent caspase-9-dependent apoptosis in human lung adenocarcinoma ASTC-a-1 cells. Mol Cell Biochem 354, 29–37 (2011). https://doi.org/10.1007/s11010-011-0802-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-011-0802-9

Keywords

Search

Navigation