Article | Published:

Bafilomycin C1 induces G0/G1 cell-cycle arrest and mitochondrial-mediated apoptosis in human hepatocellular cancer SMMC7721 cells

The Journal of Antibioticsvolume 71pages808817 (2018) | Download Citation


Bafilomycin C1, which was isolated from Streptomyces albolongus in our previous work, exhibited strong cytotoxicity against several cancer cell lines. This study aimed to evaluate its antitumor effect on human hepatocellular cancer SMMC7721 cells and the underlying mechanism in vitro and in vivo. MTT assay revealed that bafilomycin C1 retarded SMMC7721 cell growth and proliferation. Western blot and real-time qPCR analysis revealed that bafilomycin C1 caused partial G0/G1 phase cell-cycle arrest, downregulated the expression of cyclin D3, cyclin E1, CDK2, CDK4, and CDK6 and upregulated the expression of p21. Moreover, bafilomycin C1 caused mitochondrial membrane dysfunction through oxidative stress. Furthermore, bafilomycin C1 decreased the expression of Bcl-2; increased the expression of Bax, p53, and P-p53; and increased cleavage of caspase-9 and caspase-3, thereby inducing the intrinsic caspase-dependent apoptotic pathway. In vivo experiments in mice suggested that bafilomycin C1 suppressed tumor growth with few side effects. Cell-cycle arrest and induced apoptosis in tumor tissues in a mouse model treated with bafilomycin C1 were demonstrated by histological analyses, western blot and TUNEL. These findings indicate that bafilomycin C1 may be a promising candidate for hepatic cellular cancer therapy.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.


  1. 1.

    Avila MA, Berasain C, Sangro B, Prieto J. New therapies for hepatocellular carcinoma. Oncogene. 2006;25:3866–84.

  2. 2.

    Forner A, Bruix J. Ablation for hepatocellular carcinoma: is there need to have a winning technique. J Hepatol. 2010;52:310–2.

  3. 3.

    Crissien AM, Frenette C. Current management of hepatocellular carcinoma. Gastroenterol Hepatol. 2014;10:153–61.

  4. 4.

    Marquardt JU, Galle PR, Teufel A. Molecular diagnosis and therapy of hepatocellular carcinoma (HCC): an emerging field for advanced technologies. J Hepatol. 2012;56:267–75.

  5. 5.

    Ding N, et al. Bafilomycins and odoriferous sesquiterpenoids from Streptomyces albolongus isolated from Elephas maximus feces. J Nat Prod. 2016;79:799–805.

  6. 6.

    Werner G, Hagenmaier H, Drautz H, Baumgartner. A, Zähner H. Bafilomycins, a new group of marcrolide antibiotics. J Antibiot. 1984;37:110–7.

  7. 7.

    Moon SS, Hwang WH, Chung YR, Shin J. New cytotoxic bafilomycin C1-amide produced by Kitasatospora cheerisanensis. J Antibiot. 2003;56:856–61.

  8. 8.

    Carr G, et al. Bafilomycins produced in culture by Streptomyces spp. isolated from marine habitats are potent inhibitors of autophagy. J Nat Prod. 2010;73:422–7.

  9. 9.

    Muench SP, et al. PA1b inhibitor binding to subunits C and E of the vacuolar ATPase reveals its insecticidal mechanism. J Biol Chem. 2014;289:16399–408.

  10. 10.

    Su H, et al. Bafilomycin C1 exert antifungal effect through disturbing sterol biosynthesis in Candida albicans. J Antibiot. 2018;71:467–76.

  11. 11.

    Liu Y, Peterson DA, Kimura H, Schubert D. Mechanism of cellular 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction. J Neurochem. 1997;69:581–93.

  12. 12.

    Otsubo T, Akiyama Y, Yanagihara K, Yuasa Y. SOX2 is frequently downregulated in gastric cancers and inhibits cell growth through cell-cycle arrest and apoptosis. Br J Cancer. 2008;98:824–31.

  13. 13.

    Li M, et al. Bigelovin triggered apoptosis in colorectal cancer in vitro and in vivo via upregulating death receptor 5 and reactive oxidative species. Sci Rep. 2017;7:42176.

  14. 14.

    Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C. A novel assay for apoptosis. Flow cytometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled Annexin V. J Immunol Methods. 1995;184:39–51.

  15. 15.

    Simon HU, Haj-Yehia A, Levi-Schaffer F. Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis. 2000;5:415–8.

  16. 16.

    Ma J, et al. 8,9-Epoxyeicosatrienoic acid analog protects pulmonary artery smooth muscle cells from apoptosis via ROCK pathway. Exp Cell Res. 2010;316:2340–53.

  17. 17.

    Li Y, et al. Activation of sirtuin 3 by silybin attenuates mitochondrial dysfunction in cisplatin-induced acute kidney injury. Front Pharmacol. 2017;8:178.

  18. 18.

    Bi X, et al. Anti-inflammatory effects, SAR, and action mechanism of monoterpenoids from Radix Paeoniae Alba on LPS-stimulated RAW 264.7 cells. Molecules. 2017;22:e715.

  19. 19.

    Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 2001;29:e45.

  20. 20.

    Jiang X, et al. Diallyl trisulfide inhibits growth of NCI-H460 in vitro and in vivo, and ameliorates cisplatin-induced oxidative injury in the treatment of lung carcinoma in xenograft mice. Int J Biol Sci. 2017;13:167–78.

  21. 21.

    Guo ZL, et al. The novel thiosemicarbazone, di-2-pyridylketone 4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC), inhibits neuroblastoma growth in vitro and in vivo via multiple mechanisms. J Hematol Oncol. 2016;9:98.

  22. 22.

    Häcker G. The morphology of apoptosis. Cell Tissue Res. 2000;301:5–17.

  23. 23.

    Chan DC. Mitochondria: dynamic organelles in disease, aging, and development. Cell. 2006;125:1241–52.

  24. 24.

    Malumbres M, Barbacid M. Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 2009;9:153–66.

  25. 25.

    Graña X, Reddy EP. Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Oncogene. 1995;11:211–9.

  26. 26.

    Pavletich NP. Mechanisms of cyclin-dependent kinase regulation: structures of Cdks, their cyclin activators, and Cip and INK4 inhibitors. J Mol Biol. 1999;287:821–8.

  27. 27.

    Williams GH, Stoeber K. The cell cycle and cancer. J Pathol. 2012;226:352–64.

  28. 28.

    Malumbres M, Barbacid M. Mammalian cyclin-dependent kinases. Trends Biochem Sci. 2005;30:630–41.

  29. 29.

    Lundberg AS, Weinberg RA. Functional inactivation of the retinoblastoma protein requires sequential modification by at least two distinct cyclin-cdk complexes. Mol Cel Biol. 1998;18:753–61.

  30. 30.

    Harbour JW, Luo RX, Dei Santi A, Postigo AA, Dean DC. Cdk phosphorylation triggers sequential intramolecular interactions that progressively block Rb functions as cells move through G1. Cell. 1999;98:859–69.

  31. 31.

    Sherr CJ, Roberts JM. CDK inhibitors: positive and negative regulators of G1-phase progression. Genes Dev. 1999;13:1501–12.

  32. 32.

    Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007;35:495–516.

  33. 33.

    Weedon D, Searle J, Kerr JF. Apoptosis. Its nature and implications for dermatopathology. Am J Dermatopathol. 1979;1:133–44.

  34. 34.

    Hu C, et al. E Platinum, a newly synthesized platinum compound, induces autophagy via inhibiting phosphorylation of mTOR in gastric carcinoma BGC-823 cells. Toxicol Lett. 2012;210:78–86.

  35. 35.

    He. H, et al. Physalin A induces apoptosis via p53-Noxa-mediated ROS generation, and autophagy plays a protective role against apoptosis through p38-NF-κB survival pathway in A375-S2 cells. J Ethnopharmacol. 2013;148:544–55.

  36. 36.

    Gundala SR, et al. Hydroxychavicol, a betel leaf component, inhibits prostate cancer through ROS-driven DNA damage and apoptosis. Toxicol Appl Pharm. 2014;280:86–96.

  37. 37.

    Yu MO, et al. Reactive oxygen species production has a critical role in hypoxia-induced Stat3 activation and angiogenesis in human glioblastoma. J Neurooncol. 2015;125:55–63.

  38. 38.

    Tang Q, et al. Resveratrol-induced apoptosis is enhanced by inhibition of autophagy in esophageal squamous cell carcinoma. Cancer Lett. 2013;336:325–37.

  39. 39.

    Kiraz Y, Adan A, Kartal Yandim M, Baran Y. Major apoptotic mechanisms and genes involved in apoptosis. Tumor Biol. 2016;37:8471–86.

  40. 40.

    Chen T, Zheng W, Wong YS, Yang F. Mitochondria-mediated apoptosis in human breast carcinoma MCF-7 cells induces by a novel selenadiazole derivative. Biomed Pharmacother. 2008;62:77–84.

  41. 41.

    Wang F, et al. Salinomycin inhibits proliferation and induces apoptosis of human hepatocellular carcinoma cells in vitro and in vivo. PLoS ONE. 2012;7:e50638.

  42. 42.

    Fulda S. Targeting extrinsic apoptosis in cancer: challenges and opportunities. Semin Cell Dev Biol. 2015;39:84–88.

  43. 43.

    Geng YD, et al. Icariside II-induced mitochondrion and lysosome mediated apoptosis is counterbalanced by an autophagic salvage response in hepatoblastoma. Cancer Lett. 2015;366:19–31.

  44. 44.

    Taylor RC, Cullen SP, Martin SJ. Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol. 2008;9:231–41.

  45. 45.

    Cohen GM. Caspases: the executioners of apoptosis. Biochem J. 1997;326:1–16.

  46. 46.

    Rai NK, Tripathi K, Sharma D, Shukla VK. Apoptosis: a basic physiologic process in wound healing. Int J Low Extrem Wounds. 2005;4:138–44.

  47. 47.

    Amaral JD, Castro RE, Steer CJ, Rodrigues CM. P53 and the regulation of hepatocyte apoptosis: implication for disease pathogenesis. Trends Mol Med. 2009;15:531–41.

Download references


This work was funded by National Natural Science Foundation of China (Grant No. 81573327).

Author information


  1. Institute of Microbial Pharmaceuticals, College of Life and Health Sciences, Northeastern University, 110819, Shenyang, P. R. China

    • Xiaoxiao Gao
    • , Li Han
    • , Nan Ding
    • , Yu Mu
    • , Peipei Guan
    • , Caijuan Hu
    •  & Xueshi Huang


  1. Search for Xiaoxiao Gao in:

  2. Search for Li Han in:

  3. Search for Nan Ding in:

  4. Search for Yu Mu in:

  5. Search for Peipei Guan in:

  6. Search for Caijuan Hu in:

  7. Search for Xueshi Huang in:

Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding authors

Correspondence to Li Han or Xueshi Huang.

About this article

Publication history