All Title Author
Keywords Abstract

PLOS ONE  2012 

Small-Molecule Synthetic Compound Norcantharidin Reverses Multi-Drug Resistance by Regulating Sonic Hedgehog Signaling in Human Breast Cancer Cells

DOI: 10.1371/journal.pone.0037006

Full-Text   Cite this paper   Add to My Lib


Multi-drug resistance (MDR), an unfavorable factor compromising treatment efficacy of anticancer drugs, involves upregulated ATP binding cassette (ABC) transporters and activated Sonic hedgehog (Shh) signaling. By preparing human breast cancer MCF-7 cells resistant to doxorubicin (DOX), we examined the effect and mechanism of norcantharidin (NCTD), a small-molecule synthetic compound, on reversing multidrug resistance. The DOX-prepared MCF-7R cells also possessed resistance to vinorelbine, characteristic of MDR. At suboptimal concentration, NCTD significantly inhibited the viability of DOX-sensitive (MCF-7S) and DOX-resistant (MCF-7R) cells and reversed the resistance to DOX and vinorelbine. NCTD increased the intracellular accumulation of DOX in MCF-7R cells and suppressed the upregulated the mdr-1 mRNA, P-gp and BCRP protein expression, but not the MRP-1. The role of P-gp was strengthened by partial reversal of the DOX and vinorelbine resistance by cyclosporine A. NCTD treatment suppressed the upregulation of Shh expression and nuclear translocation of Gli-1, a hallmark of Shh signaling activation in the resistant clone. Furthermore, the Shh ligand upregulated the expression of P-gp and attenuated the growth inhibitory effect of NCTD. The knockdown of mdr-1 mRNA had not altered the expression of Shh and Smoothened in both MCF-7S and MCF-7R cells. This indicates that the role of Shh signaling in MDR might be upstream to mdr-1/P-gp, and similar effect was shown in breast cancer MDA-MB-231 and BT-474 cells. This study demonstrated that NCTD may overcome multidrug resistance through inhibiting Shh signaling and expression of its downstream mdr-1/P-gp expression in human breast cancer cells.


[1]  Weigel MT, Meinhold-Heerlein I, Bauerschlag DO, Schem C, Bauer M, et al. (2009) Combination of imatinib and vinorelbine enhances cell growth inhibition in breast cancer cells via PDGFR b signaling. Cancer Lett 273: 70–79.
[2]  Hines SJ, Litz JS, Krystal GW (1999) Coexpression of c-kit and stem cell factor in breast cancer results in enhanced sensitivity to members of the EGF family of growth factors. Breast Cancer Res Treat 58: 1–10.
[3]  Faneyte IF, Kristel PM, Maliepaard M, Scheffer GL, Scheper RJ, et al. (2002) Expression of the breast cancer resistance protein in breast cancer. Clin Cancer Res 8: 1068–1074.
[4]  Vasconcelos FC, Cavalcanti GB Jr, Silva KL, de Meis E, Kwee JK, et al. (2007) Contrasting features of MDR phenotype in leukemias by using two fluorochromes: implications for clinical practice. Leuk Res 31: 445–454.
[5]  Patel NH, Rothenberg ML (1994) Multidrug resistance in cancer chemotherapy. Invest New Drugs 12: 1–13.
[6]  Legrand O, Simonin G, Beauchamp-Nicoud A, Zittoun R, Marie JP (1999) Simultaneous activity of MRP1 and Pgp is correlated with in vitro resistance to daunorubicin and with in vivo resistance in adult acute myeloid leukemia. Blood 94: 1046–1056.
[7]  Jia J, Jiang J (2006) Decoding the Hedgehog signal in animal development. Cell Mol Life Sci 63: 1249–1265.
[8]  Feng YZ, Shiozawa T, Miyamoto T, Kashima H, Kurai M, et al. (2007) Overexpression of hedgehog signaling molecules and its involvement in the proliferation of endometrial carcinoma cells. Clin Cancer Res 13: 1389–1398.
[9]  Chen YJ, Wang YF, Chao KSC (2009) Cancer stem cells and sonic hedgehog signaling in head and neck cancer: potential targets for overcoming chemoradiation resistance. J Clin Oncol Soc 25: 81–88.
[10]  Chen YJ, Sims-Mourtada1 J, Izzo J, Chao KS (2007) Targeting the hedgehog pathway to mitigate treatment resistance. Cell Cycle 6: 1826–1830.
[11]  Li Y, Yang W, Yang Q, Zhou S (2012) Nuclear localization of GLI1 and elevated expression of FOXC2 in breast cancer is associated with the basal-like phenotype. Histol Histopathol 27: 475–484.
[12]  Jiao X, Wood LD, Lindman M, Jones S, Buckhaults P, et al. (2012) Somatic mutations in the notch, NF-κB, PIK3CA, and hedgehog pathways in human breast cancers. Genes Chromosomes Cancer 51: 480–489.
[13]  Chen YJ, Kuo CD, Tsai YM, Yu CC, Wang GS, et al. (2008) Norcantharidin induces anoikis through Jun-N-terminal kinase activation in CT26 colorectal cancer cells. Anticancer Drugs 19: 55–64.
[14]  Chen YJ, Tsai YM, Kuo CD, Ku KL, Shie HS, et al. (2009) Norcantharidin is a small-molecule synthetic compound with anti-angiogenesis characteristics. Life Sci 85: 642–651.
[15]  Chen YJ, Chang WM, Liu YW, Lee CY, Jang YH, et al. (2009) A small-molecule metastasis inhibitor, norcantharidin, downregulates matrix metalloproteinase-9 expression by inhibiting Sp1 transcriptional activity in colorectal cancer cells. Chem Biol Interact 181: 440–446.
[16]  Liao HF, Su SL, Chen YJ, Chou CH, Kuo CD (2007) Norcantharidin preferentially induces apoptosis in human leukemic Jurkat cells without affecting viability of normal blood mononuclear cells. Food Chem Toxicol 45: 1678–1687.
[17]  Mechetner E, Kyshtoobayeva A, Zonis S, Kim H, Stroup R, et al. (1998) Levels of multidrug resistance (MDR1) P-glycoprotein expression by human breast cancer correlate with in vitro resistance to taxol and doxorubicin. Clin Cancer Res 4: 389–398.
[18]  McCluskey A, Walkom C, Bowyer MC, Ackland SP, Gardiner E, et al. (2001) Cantharimides: a new class of modified cantharidin analogues inhibiting protein phosphatases 1 and 2A. Bioorg Med Chem Lett 11: 2941–2946.
[19]  Wang GS (1989) Medical uses of mylabris in ancient China and recent studies. J Ethnopharmacol 26: 147–162.
[20]  Chen YJ, Shieh CJ, Tsai TH, Kuo CD, Ho LT, et al. (2005) Inhibitory effect of norcantharidin, a derivative compound from blister beetles, on tumor invasion and metastasis in CT26 colorectal adenocarcinoma cells. Anticancer Drugs 16: 293–299.
[21]  Takeshita H, Kusuzaki K, Ashihara T, Gebhardt MC, Mankin HJ, et al. (2000) Intrinsic Resistance to Chemotherapeutic Agents in Murine Osteosarcoma Cells. J Bone Joint Surg Am 82-A: 963–969.
[22]  Zheng LH, Bao YL, Wu Y, Yu CL, Meng X, et al. (2008) Cantharidin reverses multidrug resistance of human gepatoma HepG2/ADM cells via down-regulation of P-glycoprotein expression. Cancer Lett 272: 102–109.
[23]  Sims-Mourtada J, Izzo JG, Ajani J, Chao KS (2007) Sonic hedgehog promotes multiple drug resistance by regulation of drug transport. Oncogene 26: 5674–5679.
[24]  Ray A, Nkhata KJ, Cleary MP (2007) Effects of leptin on human breast cancer cell lines in relationship to estrogen receptor and HER2 status. Int J Oncol 30: 1499–1509.
[25]  Lorusso V, Forcignano R, Cinieri S, Tinelli A, Porcelli L, et al. (2012) Which role for EGFR therapy in breast cancer? Front Biosci (Schol Ed) 4: 31–42.
[26]  Joshi M, Reddy SJ, Nanavidekar M, Russo JP, Russo AV, et al. (2011) Core biopsies of the breast: Diagnostic pitfalls. Indian J Pathol Microbiol 54: 671–682.
[27]  De Smaele E, Ferretti E, Gulino A (2010) Vismodegib, a small-molecule inhibitor of the hedgehog pathway for the treatment of advanced cancers. Curr Opin Investig Drugs 11: 707–718.
[28]  Gupta S, Takebe N, Lorusso P (2010) Targeting the Hedgehog pathway in cancer. Ther Adv Med Oncol 2: 237–250.
[29]  Dijkgraaf GJ, Alicke B, Weinmann L, Januario T, West K, et al. (2011) Small molecule inhibition of GDC-0449 refractory smoothened mutants and downstream mechanisms of drug resistance. Cancer Res 71: 435–444.
[30]  Carpenter RL, Lo HW (2012) Hedgehog pathway and GLI1 isoforms in human cancer. Discov Med 13: 105–113.
[31]  Yoo YA, Kang MH, Lee HJ, Kim BH, Park JK, et al. (2011) Sonic hedgehog pathway promotes metastasis and lymphangiogenesis via activation of Akt, EMT, and MMP-9 pathway in gastric cancer. Cancer Res 71: 7061–7070.
[32]  Beauchamp EM, Ringer L, Bulut G, Sajwan KP, Hall MD, et al. (2011) Arsenic trioxide inhibits human cancer cell growth and tumor development in mice by blocking Hedgehog/GLI pathway. J Clin Invest 121: 148–160.


comments powered by Disqus

Contact Us


微信:OALib Journal