Skip to main content

Advertisement

SpringerLink
Log in

Norcantharidin induces apoptosis in human prostate cancer cells through both intrinsic and extrinsic pathways

  • Original article
  • Published:
Pharmacological Reports Aims and scope Submit manuscript

Abstract

Background

Norcantharidin, a modified pure compound from blister beetles, was previously demonstrated to induce apoptosis of cancer cells. This study investigated its anti-cancer activity in prostate cancer cells and the mechanisms involved.

Methods

Two human prostate cancer cell lines, 22Rv1 and Du145, were treated with norcantharidin at concentrations ranging from 3 to 30 μg/ml. Cytotoxic effect of norcantharidin was determined by use of the 3-(4,5-dimethylthiazol-yl)-diphenyl tetrazoliumbromide (MTT) assay. The effects of apoptosis were evaluated by cell death assay, Caspase-3, -8, -9 activity and cytochrome c release. The apoptotic related protein expressions (Bcl-2 family and inhibitor of apoptosis proteins) were determined using western blotting.

Results

An MTT assay revealed that norcantharidin induced cytotoxicity against both prostate cancer cells in dose- and time-dependent manners. Treatment with norcantharidin at 3 μg/ml or higher significantly increased oligonucleosomal formation with concomitant appearance of PARP cleavage, implicating the induction of apoptosis. Norcantharidin intrinsically elevated cytosolic cytochrome c levels and activated caspase-3, -8, and -9. Extrinsically, it upregulated the expression of not only the death receptors Fas and DR5 in 22Rv1 cells, but also of RIP and TRADD adaptor proteins in Du145 cells. Mechanistically, norcantharidin increased ratios of pro-/anti-apoptotic proteins and decreased expression of IAP family member proteins, including cIAP1 and survivin, regardless of the distinct status of androgen receptor expression in both cells.

Conclusions

Norcantharidin exhibited cytotoxicity against 22Rv1 and Du145 prostate cancer cells by inducing both intrinsic and extrinsic apoptotic pathways and could thus potentially be a remedy for prostate cancer.

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

Similar content being viewed by others

References

  1. Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA Cancer J Clin 2014;64:9–29.

    Article  PubMed  Google Scholar 

  2. Ganju A, Yallapu MM, Khan S, Behrman SW, Chauhan SC, Jaggi M. Nanoways to overcome docetaxel resistance in prostate cancer. Drug Resist Updat 2014; 17: 13–23.

    Article  PubMed  PubMed Central  Google Scholar 

  3. Wang GS. Medical uses of mylabris in ancient China and recent studies. J Ethnopharmacol 1989;26:147–62.

    Article  CAS  PubMed  Google Scholar 

  4. Kok SH, Hong CY, Kuo MY, Lee CH, Lee JJ, Lou IU, et al. Comparisons of norcantharidin cytotoxic effects on oral cancer cells and normal buccal keratinocytes. Oral Oncol 2003;39:19–26.

    Article  CAS  PubMed  Google Scholar 

  5. Yang EB, Tang WY, Zhang K, Cheng LY, Mack PO. Norcantharidin inhibits growth of human HepG2 cell-transplanted tumor in nude mice and prolongs host survival. Cancer Lett 1997;117:93–8.

    Article  CAS  PubMed  Google Scholar 

  6. Chen YN, Chen JC, Yin SC, Wang GS, Tsauer W, Hsu SF, et al. Effector mechanisms of norcantharidin-induced mitotic arrest and apoptosis in human hepatoma cells. Int J Cancer 2002;100:158–65.

    Article  CAS  PubMed  Google Scholar 

  7. Yeh CH, Yang YY, Huang YF, Chow KC, Chen MF. Induction of apoptosis in human Hep3B hepatoma cells by norcantharidin through a p53 independent pathway via TRAIL/DR5 signal transduction. Chin J Integr Med 2012;18:676–82.

    Article  CAS  PubMed  Google Scholar 

  8. Zhang S, Li G, Ma X, Wang Y, Liu G, Feng L, et al. Norcantharidin enhances ABT-737-induced apoptosis in hepatocellular carcinoma cells by transcriptional repression of Mcl-1. Cell Signal 2012;24:1803–9.

    Article  CAS  PubMed  Google Scholar 

  9. Liao HF, Chen YJ, Chou CH, Wang FW, Kuo CD. Norcantharidin induces cell cycle arrest and inhibits progression of human leukemic Jurkat T cells through mitogen-activated protein kinase-mediated regulation of interleukin-2 production. Toxicol In Vitro 2011;25:206–12.

    Article  CAS  PubMed  Google Scholar 

  10. Peng F, Wei YQ, Tian L, Yang L, Zhao X, Lu Y, et al. Induction of apoptosis by norcantharidin in human colorectal carcinoma cell lines: involvement of the CD95 receptor/ligand. J Cancer Res Clin Oncol 2002;128:223–30.

    Article  CAS  PubMed  Google Scholar 

  11. Chen YJ, Kuo CD, Tsai YM, Yu CC, Wang GS, Liao HF. Norcantharidin induces anoikis through Jun-N-terminal kinase activation in CT26 colorectal cancer cells. Anticancer Drugs 2008;19:55–64.

    Article  PubMed  CAS  Google Scholar 

  12. Yang PY, Chen MF, Tsai CH, Hu DN, Chang FR, Wu YC. Involvement of caspase and MAPK activities in norcantharidin-induced colorectal cancer cell apoptosis. Toxicol In Vitro 2010;24:766–75.

    Article  CAS  PubMed  Google Scholar 

  13. An WW, Wang MW, Tashiro S, Onodera S, Ikejima T. Norcantharidin induces human melanoma A375-S2 cell apoptosis through mitochondrial and caspase pathways. J Korean Med Sci 2004;19:560–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. An WW, Wang WM, Tashiro SI, Onodera S, Ikejima T. Mitogen-activated protein kinase-dependent apoptosis in norcan-tharidin-treated A375-S2 cells is proceeded by the activation of protein kinase C. Chin Med J 2005;118:198–203.

    CAS  PubMed  Google Scholar 

  15. Hong CY, Huang SC, Lin SK, Lee JJ, Chueh LL, Lee CH, et al. Norcantharidin-induced post-G(2)/M apoptosis is dependent on wild-type p53 gene. Biochem Biophys Res Commun 2000;276:278–85.

    Article  CAS  PubMed  Google Scholar 

  16. Fan YZ, Zhao ZM, Fu JY, Chen CQ, Sun W. Norcantharidin inhibits growth of human gallbladder carcinoma xenografted tumors in nude mice by inducing apoptosis and blocking the cell cycle in vivo. Hepatobiliary Pancreat Dis Int 2010;9:414–22.

    CAS  PubMed  Google Scholar 

  17. Chen S, Wan P, Ding W, Li F, He C, Chen P, et al. Norcantharidin inhibits DNA replication and induces mitotic catastrophe by degrading initiation protein Cdc6. Int J Mol Med 2013;32:43–50.

    Article  PubMed  CAS  Google Scholar 

  18. Shen B, He PJ, Shao CL. Norcantharidin induced DU145 cell apoptosis through ROS-mediated mitochondrial dysfunction and energy depletion. PLoS One 2013;8:e84610.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  19. Xiao W, Dai B, Zhu Y, Ye D. Norcantharidin induces autophagy-related prostate cancer cell death through Beclin-1 upregulation by miR-129-5p suppression. Tumour Biol 2015;36:1–6.

    Google Scholar 

  20. Chen YJ, Shieh CJ, Tsai TH, Kuo CD, Ho LT, Liu TY, et al. Inhibitory effect of norcantharidin, a derivative compound from blister beetles, on tumor invasion and metastasis in CT26 colorectal adenocarcinoma cells. Anticancer Drugs 2005;16:293–9.

    Article  CAS  PubMed  Google Scholar 

  21. Cimmino F, Scoppettuolo MN, Carotenuto M, De Antonellis P, Dato VD, De Vita G, et al. Norcantharidin impairs medulloblastoma growth by inhibition of Wnt/beta-catenin signaling. J Neurooncol 2011;106:59–70.

    Article  PubMed  CAS  Google Scholar 

  22. Chen YJ, Tsai YM, Kuo CD, Ku KL, Shie HS, Liao HF. Norcantharidin is a small-molecule synthetic compound with anti-angiogenesis effect. Life Sci 2009;85:642–51.

    Article  CAS  PubMed  Google Scholar 

  23. An WW, Gong XF, Wang MW, Tashiro S, Onodera S, Ikejima T. Norcantharidin induces apoptosis in HeLa cells through caspase, MAPK, and mitochondrial pathways. Acta Pharmacol Sin 2004;25:1502–8.

    CAS  PubMed  Google Scholar 

  24. Chen YJ, Chang WM, Liu YW, Lee CY, Jang YH, Kuo CD, et al. A small-molecule metastasis inhibitor, norcantharidin, downregulates matrix metalloproteinase-9 expression by inhibiting Sp1 transcriptional activity in colorectal cancer cells. Chem Biol Interact 2009;181(3):440–6.

    Article  CAS  PubMed  Google Scholar 

  25. Yang PY, Chen MF, Kao YH, Hu DN, Chang FR, Wu YC. Norcantharidin induces apoptosis of breast cancer cells: involvement of activities of mitogen activated protein kinases and signal transducers and activators of transcription. Toxicol In Vitro 2011;25:699–707.

    Article  PubMed  CAS  Google Scholar 

  26. Chen YJ, Kuo CD, Chen SH, Chen WJ, Huang WC, Chao KS, et al. Small-molecule synthetic compound norcantharidin reverses multi-drug resistance by regulating Sonic hedgehog signaling in human breast cancer cells. PLoS One 2012;7:e37006.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Zhu ML, Horbinski CM, Garzotto M, Qian DZ, Beer TM, Kyprianou N. Tubulintargeting chemotherapy impairs androgen receptor activity in prostate cancer. Cancer Res 2010;70:7992–8002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Shankar S, Chen Q, Sarva K, Siddiqui I, Srivastava RK. Curcumin enhances the apoptosis-inducing potential of TRAIL in prostate cancer cells: molecular mechanisms of apoptosis, migration and angiogenesis. J Mol Signal 2007;2:10.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  29. Hahm ER, Arlotti JA, Marynowski SW, Singh SV. Honokiol, a constituent of oriental medicinal herb magnolia officinalis, inhibits growth of PC-3 xenografts in vivo in association with apoptosis induction. Clin Cancer Res 2008;14:1248–57.

    Article  CAS  PubMed  Google Scholar 

  30. Liao HF, Su SL, Chen YJ, Chou CH, Kuo CD. Norcantharidin preferentially induces apoptosis in human leukemic Jurkat cells without affecting viability of normal blood mononuclear cells. Food Chem Toxicol 2007;45:1678–87.

    Article  CAS  PubMed  Google Scholar 

  31. Chen YN, Cheng CC, Chen JC, Tsauer W, Hsu SL. Norcantharidin-induced apoptosis is via the extracellular signal-regulated kinase and c-Jun-NH2-terminal kinase signaling pathways in human hepatoma HepG2 cells. Br J Pharmacol 2003;140:461–70.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Lv H, Li Y, Du H, Fang J, Song X, Zhang J. The synthetic compound norcantharidin induced apoptosis in mantle cell lymphoma in Vivo and in vitro through the PI3K-Akt-NF-kappa B signaling pathway. Evid Based Complement Alternat Med 2013;2013:461487.

    PubMed  PubMed Central  Google Scholar 

  33. Yu CC, Ko FY, Yu CS, Lin CC, Huang YP, Yang JS, et al. Norcantharidin triggers cell death and DNA damage through S-phase arrest and ROS-modulated apoptotic pathways in TSGH 8301 human urinary bladder carcinoma cells. Int J Oncol 2012;41:1050–60.

    Article  CAS  PubMed  Google Scholar 

  34. Haupt S, Berger M, Goldberg Z, Haupt Y. Apoptosis — the p53 network. J Cell Sci 2003;116:4077–85.

    Article  CAS  PubMed  Google Scholar 

  35. Sun SY, Yue P, Shroot B, Hong WK, Lotan R. Implication of c-Myc in apoptosis induced by the retinoid CD437 in human lung carcinoma cells. Oncogene 1999;18:3894–901.

    Article  CAS  PubMed  Google Scholar 

  36. Sun SY, Yue P, Hong WK, Lotan R. Induction of Fas expression and augmentation of Fas/Fas ligand-mediated apoptosis by the synthetic retinoid CD437 in human lung cancer cells. Cancer Res 2000;60:6537–43.

    CAS  PubMed  Google Scholar 

  37. Sun SY, Yue P, Chen X, Hong WK, Lotan R. The synthetic retinoid CD437 selectively induces apoptosis in human lung cancer cells while sparing normal human lung epithelial cells. Cancer Res 2002;62:2430–6.

    CAS  PubMed  Google Scholar 

  38. Kok SH, Cheng SJ, Hong CY, Lee JJ, Lin SK, Kuo YS, et al. Norcantharidin-induced apoptosis in oral cancer cells is associated with an increase of proapoptotic to antiapoptotic protein ratio. Cancer Lett 2005;217:43–52.

    Article  CAS  PubMed  Google Scholar 

  39. Huang Y, Liu Q, Liu K, Yagasaki K, Zhang G. Suppression of growth of highly-metastatic human breast cancer cells by norcantharidin and its mechanisms of action. Cytotechnology 2009.

  40. Srinivasula SM, Ashwell JD. IAPs: what’s in a name. Mol Cell 2008;30:123–35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Kelly RJ, Lopez-Chavez A, Citrin D, Janik JE, Morris JC. Impacting tumor cell-fate by targeting the inhibitor of apoptosis protein survivin. Mol Cancer 2011;10:35.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Fan YZ, Fu JY, Zhao ZM, Chen CQ. Inhibitory effect of norcantharidin on the growth of human gallbladder carcinoma GBC-SD cells in vitro. Hepatobiliary Pancreat Dis Int 2007;6:72–80.

    CAS  PubMed  Google Scholar 

  43. Conte D, Holcik M, Lefebvre CA, Lacasse E, Picketts DJ, Wright KE, et al. Inhibitor of apoptosis protein cIAP2 is essential for lipopolysaccharide-induced macrophage survival. Mol Cell Biol 2006;26:699–708.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Tse BW, Jovanovic L, Nelson CC, de Souza P, Power CA, Russell PJ. From bench to bedside: immunotherapy for prostate cancer. Biomed Res Int 2014;2014:981434. http://dx.doi.org/10.1155/2014/981434.

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  45. Hsieh CH, Chao KS, Liao HF, Chen YJ. Norcantharidin, derivative of cantharidin, for cancer stem cells. Evid Based Complement Alternat Med 2013;2013:838651. http://dx.doi.org/10.1155/2013/838651.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Author notes

  1. These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Research, Show Chwan Memorial Hospital, Changhua, Taiwan, ROC

    Pei-Yu Yang

  2. Tissue Culture Center, New York Eye and Ear Infirmary of Mount Sinai, New York, NY, USA

    Dan-Ning Hu

  3. Department of Medical Research, E-DA Hospital, Kaohsiung, Taiwan, ROC

    Ying-Hsien Kao

  4. Department of Family Medicine, Changhua Christian Hospital, Changhua, Taiwan, ROC

    I-Ching Lin

  5. School of Medicine, Chung Shan Medical University, Taichung, Taiwan, ROC

    I-Ching Lin

  6. School of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC

    I-Ching Lin

  7. Division of Urology, Department of Surgery, Show-Chwan Memorial Hospital, Changhua, Taiwan, ROC

    Chih-Yuan Chou

  8. School of Pharmacy, College of Pharmacy, China Medical University, Taichung, Taiwan, ROC

    Yang-Chang Wu

Authors
  1. Pei-Yu Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dan-Ning Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ying-Hsien Kao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. I-Ching Lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chih-Yuan Chou
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yang-Chang Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to I-Ching Lin.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yang, PY., Hu, DN., Kao, YH. et al. Norcantharidin induces apoptosis in human prostate cancer cells through both intrinsic and extrinsic pathways. Pharmacol. Rep 68, 874–880 (2016). https://doi.org/10.1016/j.pharep.2016.04.010

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1016/j.pharep.2016.04.010

Keywords

Search

Navigation