Tumor Biology

, Volume 36, Issue 4, pp 2957–2964 | Cite as

Metformin sensitizes hepatocellular carcinoma to arsenic trioxide-induced apoptosis by downregulating Bcl2 expression

  • Xuejun Yang
  • Deguang Sun
  • Yu Tian
  • Sunbin Ling
  • Liming WangEmail author
Research Article


Hepatocellular carcinoma (HCC) is a highly malignant tumor that can evolve rapidly to acquire resistance to conventional chemotherapies. Arsenic trioxide (ATO) is a traditional Asian medicine, and a phase II study has shown that treatment with ATO alone was not effective against HCC. Bcl2 is an antiapoptotic protein that regulates chemotherapy in HCC. Metformin is reported to decrease Bcl2 expression, and the purpose of this study was to verify whether metformin could potentiate the anti-HCC efficacy of ATO in vitro. In the present study, we used metformin and ATO alone or in combination and then tested proliferation, apoptosis, and Bcl2 level of HCC cells. The results showed that metformin enhanced both the proliferation-inhibiting and apoptosis-inducing effects of ATO on HCC cell lines HepG2 and BEL7402. Furthermore, this activity proceeded via a mechanism involving metformin-induced downregulation of Bcl2. A combination of ATO and metformin is therefore a potentially promising approach for HCC therapy.


Hepatocellular carcinoma Apoptosis Bcl2 Metformin Arsenic trioxide 



The authors appreciate Professor Hanfa Zou and Professor Mingliang Ye for their technical support in the research. This research was supported by the National Nature Science Foundation of China (No. 81272368).

Conflicts of interest



  1. 1.
    Zheng Y, Wang X, Wang H, Yan W, Zhang Q, Chang X. Bone morphogenetic protein 2 inhibits hepatocellular carcinoma growth and migration through downregulation of the PI3K/AKT pathway. Tumour Biol. 2014;35(6):5189–98.CrossRefPubMedGoogle Scholar
  2. 2.
    Yao M, Wang L, Dong Z, Qian Q, Shi Y, Yu D, et al. Glypican-3 as an emerging molecular target for hepatocellular carcinoma gene therapy. Tumour Biol. 2014;35(6):5857–68.CrossRefPubMedGoogle Scholar
  3. 3.
    Yao N, Yao D, Wang L, Dong Z, Wu W, Qiu L, et al. Inhibition of autocrine IGF-II on effect of human HepG2 cell proliferation and angiogenesis factor expression. Tumour Biol. 2012;33(5):1767–76.CrossRefPubMedGoogle Scholar
  4. 4.
    Zheng L, Gong W, Liang P, Huang X, You N, Han KQ, et al. Effects of AFP-activated PI3K/Akt signaling pathway on cell proliferation of liver cancer. Tumour Biol. 2014;35(5):4095–9.CrossRefPubMedGoogle Scholar
  5. 5.
    Liu ZM, Tseng JT, Hong DY, Huang HS. Suppression of TG-interacting factor sensitizes arsenic trioxide-induced apoptosis in human hepatocellular carcinoma cells. Biochem J. 2011;438(2):349–58.CrossRefPubMedGoogle Scholar
  6. 6.
    Antman KH. Introduction: the history of arsenic trioxide in cancer therapy. Oncologist. 2001;6 Suppl 2:1–2.CrossRefPubMedGoogle Scholar
  7. 7.
    Park MT, Kang YH, Park IC, Kim CH, Lee YS, Chung HY, et al. Combination treatment with arsenic trioxide and phytosphingosine enhances apoptotic cell death in arsenic trioxide-resistant cancer cells. Mol Cancer Ther. 2007;6(1):82–92.CrossRefPubMedGoogle Scholar
  8. 8.
    Beauchamp EM, Ringer L, Bulut G, Sajwan KP, Hall MD, Lee YC, et al. Arsenic trioxide inhibits human cancer cell growth and tumor development in mice by blocking Hedgehog/GLI pathway. J Clin Invest. 2011;121(1):148–60.CrossRefPubMedGoogle Scholar
  9. 9.
    Ma Y, Wang J, Liu L, Zhu H, Chen X, Pan S, et al. Genistein potentiates the effect of arsenic trioxide against human hepatocellular carcinoma: role of Akt and nuclear factor-kappaB. Cancer Lett. 2011;301(1):75–84.CrossRefPubMedGoogle Scholar
  10. 10.
    Lin CC, Hsu C, Hsu CH, Hsu WL, Cheng AL, Yang CH. Arsenic trioxide in patients with hepatocellular carcinoma: a phase II trial. Invest New Drugs. 2007;25(1):77–84.CrossRefPubMedGoogle Scholar
  11. 11.
    Takahashi A, Kimura F, Yamanaka A, Takebayashi A, Kita N, Takahashi K, et al. Metformin impairs growth of endometrial cancer cells via cell cycle arrest and concomitant autophagy and apoptosis. Cancer Cell Int. 2014;14:53.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Feng Y, Ke C, Tang Q, Dong H, Zheng X, Lin W, et al. Metformin promotes autophagy and apoptosis in esophageal squamous cell carcinoma by downregulating Stat3 signaling. Cell Death Dis. 2014;5:e1088.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Xiao Y, Zhang S, Hou G, Zhang X, Hao X, Zhang J. Clinical pathological characteristics and prognostic analysis of diabetic women with luminal subtype breast cancer. Tumour Biol. 2014;35(3):2035–45.CrossRefPubMedGoogle Scholar
  14. 14.
    Rocha GZ, Dias MM, Ropelle ER, Osorio-Costa F, Rossato FA, Vercesi AE, et al. Metformin amplifies chemotherapy-induced AMPK activation and antitumoral growth. Clin Cancer Res. 2011;17(12):3993–4005.CrossRefPubMedGoogle Scholar
  15. 15.
    Ben Sahra I, Laurent K, Loubat A, Giorgetti-Peraldi S, Colosetti P, Auberger P, et al. The antidiabetic drug metformin exerts an antitumoral effect in vitro and in vivo through a decrease of cyclin D1 level. Oncogene. 2008;27(25):3576–86.CrossRefPubMedGoogle Scholar
  16. 16.
    Isakovic A, Harhaji L, Stevanovic D, Markovic Z, Sumarac-Dumanovic M, Starcevic V, et al. Dual antiglioma action of metformin: cell cycle arrest and mitochondria-dependent apoptosis. Cell Mol Life Sci. 2007;64(10):1290–302.CrossRefPubMedGoogle Scholar
  17. 17.
    Zakikhani M, Dowling RJ, Sonenberg N, Pollak MN. The effects of adiponectin and metformin on prostate and colon neoplasia involve activation of AMP-activated protein kinase. Cancer Prev Res (Phila). 2008;1(5):369–75.CrossRefGoogle Scholar
  18. 18.
    Saito T, Chiba T, Yuki K, Zen Y, Oshima M, Koide S, et al. Metformin, a diabetes drug, eliminates tumor-initiating hepatocellular carcinoma cells. PLoS One. 2013;8(7):e70010.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Do MT, Kim HG, Khanal T, Choi JH, Kim DH, Jeong TC, et al. Metformin inhibits heme oxygenase-1 expression in cancer cells through inactivation of Raf-ERK-Nrf2 signaling and AMPK-independent pathways. Toxicol Appl Pharmacol. 2013;271(2):229–38.CrossRefPubMedGoogle Scholar
  20. 20.
    Bodmer M, Meier C, Krahenbuhl S, Jick SS, Meier CR. Long-term metformin use is associated with decreased risk of breast cancer. Diabetes Care. 2010;33(6):1304–8.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Jiralerspong S, Palla SL, Giordano SH, Meric-Bernstam F, Liedtke C, Barnett CM, et al. Metformin and pathologic complete responses to neoadjuvant chemotherapy in diabetic patients with breast cancer. J Clin Oncol. 2009;27(20):3297–302.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Zeng HZ, Qu YQ, Zhang WJ, Xiu B, Deng AM, Liang AB. Proteomic analysis identified DJ-1 as a cisplatin resistant marker in non-small cell lung cancer. Int J Mol Sci. 2011;12:3489–99.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Han C, Gu H, Wang J, Lu W, Mei Y, Wu M. Regulation of L-threonine dehydrogenase in somatic cell reprogramming. Stem Cells. 2013;31(5):953–65.CrossRefPubMedGoogle Scholar
  24. 24.
    Yang CL, Jiang FQ, Xu F, Jiang GX. ADAM10 overexpression confers resistance to doxorubicin-induced apoptosis in hepatocellular carcinoma. Tumour Biol. 2012;33(5):1535–41.CrossRefPubMedGoogle Scholar
  25. 25.
    Davison K, Mann KK, Miller Jr WH. Arsenic trioxide: mechanisms of action. Semin Hematol. 2002;39(2 Suppl 1):3–7.CrossRefPubMedGoogle Scholar
  26. 26.
    Chen GQ, Zhu J, Shi XG, Ni JH, Zhong HJ, Si GY, et al. In vitro studies on cellular and molecular mechanisms of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia: As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML proteins. Blood. 1996;88(3):1052–61.PubMedGoogle Scholar
  27. 27.
    Oketani M, Kohara K, Tuvdendorj D, Ishitsuka K, Komorizono Y, Ishibashi K, et al. Inhibition by arsenic trioxide of human hepatoma cell growth. Cancer Lett. 2002;183(2):147–53.CrossRefPubMedGoogle Scholar
  28. 28.
    Siu KP, Chan JY, Fung KP. Effect of arsenic trioxide on human hepatocellular carcinoma HepG2 cells: inhibition of proliferation and induction of apoptosis. Life Sci. 2002;71(3):275–85.CrossRefPubMedGoogle Scholar
  29. 29.
    Chaudhary SC, Kurundkar D, Elmets CA, Kopelovich L, Athar M. Metformin, an antidiabetic agent reduces growth of cutaneous squamous cell carcinoma by targeting mTOR signaling pathway. Photochem Photobiol. 2012;88(5):1149–56.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Carmignani M, Volpe AR, Aldea M, Soritau O, Irimie A, Florian IS, et al. Glioblastoma stem cells: a new target for metformin and arsenic trioxide. J Biol Regul Homeost Agents. 2014;28(1):1–15.PubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Xuejun Yang
    • 1
    • 2
  • Deguang Sun
    • 2
  • Yu Tian
    • 1
  • Sunbin Ling
    • 2
    • 3
  • Liming Wang
    • 2
    Email author
  1. 1.Dalian Medical UniversityDalianChina
  2. 2.Department of General SurgeryThe Second Affiliated Hospital of Dalian Medical UniversityDalianChina
  3. 3.Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated HospitalZhejiang University School of MedicineHangzhouChina

Personalised recommendations