Tumor Biology

, Volume 33, Issue 5, pp 1535–1541 | Cite as

ADAM10 overexpression confers resistance to doxorubicin-induced apoptosis in hepatocellular carcinoma

  • Cheng-lin Yang
  • Feng-qin Jiang
  • Feng Xu
  • Gui-xing Jiang
Research Article

Abstract

Chemoresistance represents a major obstacle to successful treatment of hepatocellular carcinoma (HCC). A disintegrin and metalloproteinase 10 (ADAM10) is known to be frequently upregulated in many cancers. We aimed to determine the biological function of ADAM10 in the chemoresistance of HCC cells. Overexpression of ADAM10 in three HCC cell lines (HepG2, Hep3B, and Huh7) conferred protection against doxorubicin-induced apoptosis, as determined by Annexin V staining. Western blot analysis revealed that ADAM10-overexpressing cells had a significantly lower amount of cleaved caspase-3 and an elevated expression of myeloid cell leukemia-1 (Mcl-1), a prosurvival member of the Bcl-2 family. Conversely, RNA interference-mediated silencing of endogenous ADAM10 potentiated doxorubicin-induced apoptosis in HepG2 and Hep3B cells, which was coupled with increased cleavage of caspase-3 and decreased expression of Mcl-1. Ectopic expression of ADAM10 resulted in a marked increase in the phosphorylation of phosphatidylinositol 3-kinase (PI3-K) and Akt. Most interestingly, the pretreatment with the PI3-K inhibitor LY294002 significantly enhanced doxorubicin-induced apoptosis and diminished the Mcl-1 expression in ADAM10-overexpressing Huh7 cells. Our data provide evidence that ADAM10 plays an important role in modulating the chemosensitivity of HCC cells, which, at least partially, involves the activation of the PI3-K/Akt pathway. ADAM10 may be a promising target for the improvement of chemotherapeutic efficacy in HCC.

Keywords

Hepatocellular carcinoma (HCC) ADAM10 Chemoresistance Apoptosis Akt 

Supplementary material

13277_2012_405_MOESM1_ESM.doc (34 kb)
Supplementary Fig S1Hep3B cells were exposed to doxorubicin at concentrations of 0.1 to 5.0 μM for 24 h, and cell apoptosis was assessed by flow cytometry using the Annexin V-FITC Apoptosis Detection Kit. Doxorubicin at the concentration of 5.0 μM had a maximal induction of apoptosis. Error bars indicate standard deviations from 3 independent experiments. (DOC 34 kb)

References

  1. 1.
    Parkin DM, Bray F, Ferlay J, Pisani P. Global cancer statistics, 2002. CA Cancer J Clin. 2005;55:74–108.CrossRefPubMedGoogle Scholar
  2. 2.
    Chung GE, Lee JH, Kim HY, et al. Transarterial chemoembolization can be safely performed in patients with hepatocellular carcinoma invading the main portal vein and may improve the overall survival. Radiology. 2011;258:627–34.CrossRefPubMedGoogle Scholar
  3. 3.
    Yoo DJ, Kim KM, Jin YJ, et al. Clinical outcome of 251 patients with extrahepatic metastasis at initial diagnosis of hepatocellular carcinoma: does transarterial chemoembolization improve survival in these patients? J Gastroenterol Hepatol. 2011;26:145–54.CrossRefPubMedGoogle Scholar
  4. 4.
    Dhanasekaran R, Kooby DA, Staley CA, Kauh JS, Khanna V, Kim HS. Comparison of conventional transarterial chemoembolization (TACE) and chemoembolization with doxorubicin drug eluting beads (DEB) for unresectable hepatocelluar carcinoma (HCC). J Surg Oncol. 2010;101:476–80.PubMedGoogle Scholar
  5. 5.
    Song MJ, Park CH, Kim JD, et al. Drug-eluting bead loaded with doxorubicin versus conventional Lipiodol-based transarterial chemoembolization in the treatment of hepatocellular carcinoma: a case-control study of Asian patients. Eur J Gastroenterol Hepatol. 2011;23:521–7.CrossRefPubMedGoogle Scholar
  6. 6.
    Seals DF, Courtneidge SA. The ADAMs family of metalloproteases: multidomain proteins with multiple functions. Genes Dev. 2003;17:7–30.CrossRefPubMedGoogle Scholar
  7. 7.
    Klein T, Bischoff R. Active metalloproteases of the A disintegrin and metalloprotease (ADAM) family: biological function and structure. J Proteome Res. 2011;10:17–33.CrossRefPubMedGoogle Scholar
  8. 8.
    Wang YY, Ye ZY, Li L, Zhao ZS, Shao QS, Tao HQ. ADAM 10 is associated with gastric cancer progression and prognosis of patients. J Surg Oncol. 2011;103:116–23.CrossRefPubMedGoogle Scholar
  9. 9.
    Gaida MM, Haag N, Günther F, et al. Expression of A disintegrin and metalloprotease 10 in pancreatic carcinoma. Int J Mol Med. 2010;26:281–8.PubMedGoogle Scholar
  10. 10.
    Lee SB, Schramme A, Doberstein K, et al. ADAM10 is upregulated in melanoma metastasis compared with primary melanoma. J Invest Dermatol. 2010;130:763–73.CrossRefPubMedGoogle Scholar
  11. 11.
    Kohga K, Takehara T, Tatsumi T, et al. Anticancer chemotherapy inhibits MHC class I-related chain a ectodomain shedding by downregulating ADAM10 expression in hepatocellular carcinoma. Cancer Res. 2009;69:8050–7.CrossRefPubMedGoogle Scholar
  12. 12.
    Gavert N, Sheffer M, Raveh S, et al. Expression of L1-CAM and ADAM10 in human colon cancer cells induces metastasis. Cancer Res. 2007;67:7703–12.CrossRefPubMedGoogle Scholar
  13. 13.
    Xu Q, Liu X, Chen W, Zhang Z. Inhibiting adenoid cystic carcinoma cells growth and metastasis by blocking the expression of ADAM 10 using RNA interference. J Transl Med. 2010;8:136.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Kirkin V, Cahuzac N, Guardiola-Serrano F, et al. The Fas ligand intracellular domain is released by ADAM10 and SPPL2a cleavage in T-cells. Cell Death Differ. 2007;14:1678–87.CrossRefPubMedGoogle Scholar
  15. 15.
    Moss ML, Bomar M, Liu Q, et al. The ADAM10 prodomain is a specific inhibitor of ADAM10 proteolytic activity and inhibits cellular shedding events. J Biol Chem. 2007;282:35712–21.CrossRefPubMedGoogle Scholar
  16. 16.
    Porter AG, Jänicke RU. Emerging roles of caspase-3 in apoptosis. Cell Death Differ. 1999;6:99–104.CrossRefPubMedGoogle Scholar
  17. 17.
    Townsend KJ, Trusty JL, Traupman MA, Eastman A, Craig RW. Expression of the antiapoptotic MCL1 gene product is regulated by a mitogen activated protein kinase-mediated pathway triggered through microtubule disruption and protein kinase C. Oncogene. 1998;17:1223–34.CrossRefPubMedGoogle Scholar
  18. 18.
    Gagnon V, Van Themsche C, Turner S, Leblanc V, Asselin E. Akt and XIAP regulate the sensitivity of human uterine cancer cells to cisplatin, doxorubicin and Taxol. Apoptosis. 2008;13:259–71.CrossRefPubMedGoogle Scholar
  19. 19.
    McDonald GT, Sullivan R, Paré GC, Graham CH. Inhibition of phosphatidylinositol 3-kinase promotes tumor cell resistance to chemotherapeutic agents via a mechanism involving delay in cell cycle progression. Exp Cell Res. 2010;316:3197–206.CrossRefPubMedGoogle Scholar
  20. 20.
    Chekenya M, Krakstad C, Svendsen A, et al. The progenitor cell marker NG2/MPG promotes chemoresistance by activation of integrin-dependent PI3K/Akt signaling. Oncogene. 2008;27:5182–94.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Kyula JN, Van Schaeybroeck S, Doherty J, Fenning CS, Longley DB, Johnston PG. Chemotherapy-induced activation of ADAM-17: a novel mechanism of drug resistance in colorectal cancer. Clin Cancer Res. 2010;16:3378–89.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Böhm B, Hess S, Krause K, et al. ADAM15 exerts an antiapoptotic effect on osteoarthritic chondrocytes via up-regulation of the X-linked inhibitor of apoptosis. Arthritis Rheum. 2010;62:1372–82.CrossRefPubMedGoogle Scholar
  23. 23.
    Rocks N, Estrella C, Paulissen G, et al. The metalloproteinase ADAM-12 regulates bronchial epithelial cell proliferation and apoptosis. Cell Prolif. 2008;41:988–1001.CrossRefPubMedGoogle Scholar
  24. 24.
    Kveiborg M, Fröhlich C, Albrechtsen R, et al. A role for ADAM12 in breast tumor progression and stromal cell apoptosis. Cancer Res. 2005;65:4754–61.CrossRefPubMedGoogle Scholar
  25. 25.
    Bai S, Nasser MW, Wang B, et al. MicroRNA-122 inhibits tumorigenic properties of hepatocellular carcinoma cells and sensitizes these cells to sorafenib. J Biol Chem. 2009;284:32015–27.CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Llovet JM, Ricci S, Mazzaferro V, et al. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 2008;359:378–90.CrossRefPubMedGoogle Scholar
  27. 27.
    Schulze-Bergkamen H, Fleischer B, Schuchmann M, et al. Suppression of Mcl-1 via RNA interference sensitizes human hepatocellular carcinoma cells towards apoptosis induction. BMC Cancer. 2006;6:232.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Hu L, Chen L, Yang G, et al. HBx sensitizes cells to oxidative stress-induced apoptosis by accelerating the loss of Mcl-1 protein via caspase-3 cascade. Mol Cancer. 2011;10:43.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Qi F, Inagaki Y, Gao B, et al. Bufalin and cinobufagin induce apoptosis of human hepatocellular carcinoma cells via Fas- and mitochondria-mediated pathways. Cancer Sci. 2011;102:951–8.CrossRefPubMedGoogle Scholar
  30. 30.
    Seitz SJ, Schleithoff ES, Koch A, et al. Chemotherapy-induced apoptosis in hepatocellular carcinoma involves the p53 family and is mediated via the extrinsic and the intrinsic pathway. Int J Cancer. 2010;126:2049–66.PubMedGoogle Scholar
  31. 31.
    Wirth T, Kühnel F, Fleischmann-Mundt B, et al. Telomerase-dependent virotherapy overcomes resistance of hepatocellular carcinomas against chemotherapy and tumor necrosis factor-related apoptosis-inducing ligand by elimination of Mcl-1. Cancer Res. 2005;65:7393–402.CrossRefPubMedGoogle Scholar
  32. 32.
    Allagnat F, Cunha D, Moore F, Vanderwinden JM, Eizirik DL, Cardozo AK. Mcl-1 downregulation by pro-inflammatory cytokines and palmitate is an early event contributing to β-cell apoptosis. Cell Death Differ. 2011;18:328–37.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Chen KF, Chen HL, Tai WT, et al. Activation of phosphatidylinositol 3-kinase/Akt signaling pathway mediates acquired resistance to sorafenib in hepatocellular carcinoma cells. J Pharmacol Exp Ther. 2011;337:155–61.CrossRefPubMedGoogle Scholar
  34. 34.
    Chen KF, Yeh PY, Hsu C, et al. Bortezomib overcomes tumor necrosis factor-related apoptosis-inducing ligand resistance in hepatocellular carcinoma cells in part through the inhibition of the phosphatidylinositol 3-kinase/Akt pathway. J Biol Chem. 2009;284:11121–33.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Chen W, Bai L, Wang X, Xu S, Belinsky SA, Lin Y. Acquired activation of the Akt/cyclooxygenase-2/Mcl-1 pathway renders lung cancer cells resistant to apoptosis. Mol Pharmacol. 2010;77:416–23.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Wang X, Chen W, Zeng W, et al. Akt-mediated eminent expression of c-FLIP and Mcl-1 confers acquired resistance to TRAIL-induced cytotoxicity to lung cancer cells. Mol Cancer Ther. 2008;7:1156–63.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Fleischer B, Schulze-Bergkamen H, Schuchmann M, et al. Mcl-1 is an anti-apoptotic factor for human hepatocellular carcinoma. Int J Oncol. 2006;28:25–32.PubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2012

Authors and Affiliations

  • Cheng-lin Yang
    • 1
    • 2
  • Feng-qin Jiang
    • 3
  • Feng Xu
    • 3
  • Gui-xing Jiang
    • 4
  1. 1.Ankang Central HospitalAnkangChina
  2. 2.Department of General SurgeryAffiliated Hospital of Ankang Vocational and Technical CollegeAnkangChina
  3. 3.Shaoxing People’s HospitalShaoxing Hospital of Zhejiang UniversityShaoxingChina
  4. 4.Department of Surgery, Second Affiliated Hospital, School of MedicineZhejiang UniversityHangzhouChina

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