Asian Pacific Journal of Cancer Prevention

Asian Pacific Journal of Cancer Prevention (아시아태평양암예방학회)
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Blocking Bcl-2 Leads to Autophagy Activation and Cell Death of the HEPG2 Liver Cancer Cell Line

  • Du, Peng (Department of General Surgery, the Second Affiliated Hospital of Soochow University) ;
  • Cao, Hua (Department of General Surgery, the Second Affiliated Hospital of Soochow University) ;
  • Wu, Hao-Rong (Department of General Surgery, the Second Affiliated Hospital of Soochow University) ;
  • Zhu, Bao-Song (Department of General Surgery, the Second Affiliated Hospital of Soochow University) ;
  • Wang, Hao-Wei (Department of General Surgery, the Second Affiliated Hospital of Soochow University) ;
  • Gu, Chun-Wei (Department of General Surgery, the Second Affiliated Hospital of Soochow University) ;
  • Xing, Chun-Gen (Department of General Surgery, the Second Affiliated Hospital of Soochow University) ;
  • Chen, Wei (Department of General Surgery, the Second Affiliated Hospital of Soochow University)
  • Published : 2013.10.30
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Abstract

Background: Apoptosis may be induced after Bcl-2 expression is inhibited in proliferative cancer cells. This study focused on the effect of autophagy activation by ABT737 on anti-tumor effects of epirubicin. Methods: Cytotoxic effects of ABT737 on the HepG2 liver cancer cell line were assessed by MTT assay and cell apoptosis through flow cytometry. Mitochondrial membrane potential was measured by fluorescence microscopy. Monodansylcadaverin (MDC) staining was used to detect activation of autophagy. Expression of p53, p62, LC3, and Beclin1, apoptotic or autophagy related proteins, was detected by Western blotting. Results: ABT737 and epirubicin induced growth inhibition in HepG2 cells in a dose- and time-dependent manner. Both ABT737 and epirubicin alone could induce cell apoptosis with a reduction in mitochondrial membrane potential as well as increased apoptotic protein expression. Further increase of apoptosis was detected when HepG2 cells were co-treated with ABT373 and epirubicin. Furthermore, our results demonstrated that ABT373 or epirubicin ccould activate cell autophagy with elevated autophagosome formation, increased expression of autophagy related proteins and LC3 fluorescent puncta. Conclusions: ABT737 influences cancer cells through both apoptotic and autophagic mechanisms, and ABT737 may enhance the effects of epirubicin on HepG2 cells by activating autophagy and inducing apoptosis.

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References

  1. Abou-Alfa GK, Venook AP (2008). The impact of new data in the treatment of advanced hepatocellular carcinoma. Curr Oncol Rep, 10, 199-205. https://doi.org/10.1007/s11912-008-0031-x
  2. Al-Ejeh F, Kumar R, Wiegmans A, et al (2010). Harnessing the complexity of DNAdamage response pathways to improve cancer treatment outcomes. Oncogene, 29, 6085-98. https://doi.org/10.1038/onc.2010.407
  3. Alvarez-Tejado M, Naranjo-Suarez S, Jimenez C, et al (2001). Hypoxia induced the activation of the phosphatidylinositol 3-kinase/Akt cell survival pathway in PC12 cells. J Biol Chem, 276, 22368-74. https://doi.org/10.1074/jbc.M011688200
  4. Biederbick A, Kern HF, Elsasser HP (1995). Monodansylcadaverine (MDC) is a specific in vivo marker for autophagic vacuoles. Eur J Cell Biol, 66, 3-14.
  5. Bursch W, Ellinger A, Kienzl H, et al (1996). Active cell death induced by the anti-estrogens tamoxifen and ICI 164 384 in human mammary carcinoma cells (MCF-7) in culture: the role of autophagy. Carcinogenesis, 17, 1595-607. https://doi.org/10.1093/carcin/17.8.1595
  6. Buytaert E, Callewaert G, Vandenheede JR, Agostinis P (2006). Deficiency in apoptotic effectors Bax and Bak reveals an autophagic cell death pathway initiated by photodamage to the endoplasmic reticulum. Autophagy, 2, 238-40. https://doi.org/10.4161/auto.2730
  7. Cadwell K, Liu JY, Brown SL, et al (2008). A key role for autophagy and the autophagy gene Atg16l1 in mouse and human intestinal Paneth cells. Nature, 456, 259-63. https://doi.org/10.1038/nature07416
  8. Carew JS, Nawrocki ST, Cleveland JL (2007). Modulating autophagy for therapeutic benefit. Autophagy, 3, 464-7. https://doi.org/10.4161/auto.4311
  9. Chen N, Karantza-Wadsworth V (2009). Role and regulation of autophagy in cancer. Biochim Biophys Acta, 1793, 1516-23. https://doi.org/10.1016/j.bbamcr.2008.12.013
  10. Erlich S, Mizrachy L, Segev O, et al (2007). Differential interactions between Beclin 1 and Bcl-2 family members. Autophagy, 3, 561-8. https://doi.org/10.4161/auto.4713
  11. Gewirtz DA (2007). Autophagy as a mechanism of radiation sensitization in breast tumor cells. Autophagy, 3, 249-50. https://doi.org/10.4161/auto.3723
  12. Gozuacik D, Kimchi A (2004). Autophagy as a cell death and tumor suppressor mechanism. Oncogene, 23, 2891-906. https://doi.org/10.1038/sj.onc.1207521
  13. Hanahan D, Weinberg RA (2000). The hallmarks of cancer. Cell, 100, 57-70. https://doi.org/10.1016/S0092-8674(00)81683-9
  14. John S, Nayvelt I, Hsu HC, et al (2008). Regulation of estrogenic effects by beclin 1 in breast cancer cells. Cancer Res, 68, 7855-63. https://doi.org/10.1158/0008-5472.CAN-07-5875
  15. Kanzawa T, Germano IM, Komata T, et al (2004). Role of autophagy in temozolomide-inducedcytotoxicity for malignant glioma cells. Cell Death Differ, 11, 448-57. https://doi.org/10.1038/sj.cdd.4401359
  16. Karantza-Wadsworth V, White E (2008). Programmed cell death. In: DeVita, VT.; Lawrence, TS.; Rosenberg, SA., editors. Cancer: principles and practice of oncology. 8th ed. Lippincott, Williams, & Wilkins; Baltimore, pp 93-101.
  17. Kirkin V, McEwan DG, Novak I, Dikic I (2009). A role for ubiquitin in selective autophagy. Mol Cell, 34, 259-69. https://doi.org/10.1016/j.molcel.2009.04.026
  18. Lum JJ, DeBerardinis RJ, Thompson CB (2005). Thompson, Autophagy in metazoans: cell survival in the land of plenty. Nat Rev Mol Cell Biol, 6, 439-48. https://doi.org/10.1038/nrm1660
  19. Malicdan MC, Noguchi S, Nonaka I, Saftig P, Nishino I (2008). Lysosomal myopathies: an excessive build-up in autophagosomes is too much to handle. Neuromuscul Disord, 18, 521-9. https://doi.org/10.1016/j.nmd.2008.04.010
  20. Mathew R, Karantza-Wadsworth V, White E (2007). Role of autophagy in cancer. Nat Rev Cancer, 7, 961-7. https://doi.org/10.1038/nrc2254
  21. Niemann A, Takatsuki A, Elsasser HP (2000). The lysosomotropic agent monodansylcadaverine also acts as a solvent polarity probe. J Histochem Cytochem, 48, 251-8. https://doi.org/10.1177/002215540004800210
  22. Orvedahl A, Levine B (2009). Eating the enemy within: autophagy in infectious diseases. Cell Death Differ, 16, 57-69. https://doi.org/10.1038/cdd.2008.130
  23. Pankiv S, Clausen TH, Lamark T, et al (2007). p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem, 282, 24131-45. https://doi.org/10.1074/jbc.M702824200
  24. Pattingre S, Tassa A, Qu X, Garuti R, et al (2005). Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell, 122, 927-39. https://doi.org/10.1016/j.cell.2005.07.002
  25. Poon D, Anderson BO, Chen LT, et al (2009). Management of hepatocellular carcinoma in Asia: consensus statement from the Asian Oncology Summit 2009. Lancet Oncol, 10, 111l-8. https://doi.org/10.1016/S1470-2045(09)70241-4
  26. Rashid SF, Moore JS, Walker E, et al (2001). Synergistic growth inhibition of prostate cancer cells by 1 alpha, 25 Dihydroxyvitamin D(3) and its 19-nor-hexafluoride analogs in combination with either sodium butyrate or trichostatin A. Oncogene, 15, 1860-72.
  27. Saitoh T, Fujita N, Jang MH, et al (2008). Loss of the autophagy protein Atg16L1 enhances endotoxin-induced IL-1beta production. Nature, 456, 264-8. https://doi.org/10.1038/nature07383
  28. Vero V, Racco S, Biolato M, et al (2009). The treatment of hepatocellular carcinoma: an update. Minerva Med, 100, 173-93.
  29. Wei Y, Pattingre S, Sinha S, Bassik M, Levine B (2008). JNK1-mediated phosphorylation of Bcl-2 regulates starvationinduced autophagy. Mol Cell, 30, 678-88. https://doi.org/10.1016/j.molcel.2008.06.001
  30. Winslow AR, Rubinsztein DC (2008). Autophagy in neurodegeneration and development. Biochim Biophys Acta, 1782, 723-9. https://doi.org/10.1016/j.bbadis.2008.06.010
  31. Wong N, Yeo W, Wong WL, et al (2009). TOP2A overexpression in hepatocellular carcinoma correlates with early age onset, shorter patients survival and chemoresistance. Int J Cancer, 124, 644-52. https://doi.org/10.1002/ijc.23968
  32. Yen WL, Klionsky DJ (2008). How to live long and prosper:autophagy, mitochondria and aging. Physiology (Bethesda), 23, 248-62. https://doi.org/10.1152/physiol.00013.2008

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