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http://dx.doi.org/10.7314/APJCP.2013.14.10.5849

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)
Publication Information
Asian Pacific Journal of Cancer Prevention / v.14, no.10, 2013 , pp. 5849-5854 More about this Journal
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.
Keywords
Bcl-2; ABT737; autophagy; chemotherapy;
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1 Kirkin V, McEwan DG, Novak I, Dikic I (2009). A role for ubiquitin in selective autophagy. Mol Cell, 34, 259-69.   DOI   ScienceOn
2 Niemann A, Takatsuki A, Elsasser HP (2000). The lysosomotropic agent monodansylcadaverine also acts as a solvent polarity probe. J Histochem Cytochem, 48, 251-8.   DOI   ScienceOn
3 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.   DOI   ScienceOn
4 Mathew R, Karantza-Wadsworth V, White E (2007). Role of autophagy in cancer. Nat Rev Cancer, 7, 961-7.   DOI   ScienceOn
5 Orvedahl A, Levine B (2009). Eating the enemy within: autophagy in infectious diseases. Cell Death Differ, 16, 57-69.   DOI   ScienceOn
6 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.   DOI   ScienceOn
7 Pattingre S, Tassa A, Qu X, Garuti R, et al (2005). Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell, 122, 927-39.   DOI   ScienceOn
8 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.   DOI   ScienceOn
9 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.
10 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.   DOI   ScienceOn
11 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.   DOI   ScienceOn
12 Vero V, Racco S, Biolato M, et al (2009). The treatment of hepatocellular carcinoma: an update. Minerva Med, 100, 173-93.
13 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.   DOI   ScienceOn
14 Winslow AR, Rubinsztein DC (2008). Autophagy in neurodegeneration and development. Biochim Biophys Acta, 1782, 723-9.   DOI   ScienceOn
15 Yen WL, Klionsky DJ (2008). How to live long and prosper:autophagy, mitochondria and aging. Physiology (Bethesda), 23, 248-62.   DOI   ScienceOn
16 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.
17 Abou-Alfa GK, Venook AP (2008). The impact of new data in the treatment of advanced hepatocellular carcinoma. Curr Oncol Rep, 10, 199-205.   DOI   ScienceOn
18 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.   DOI   ScienceOn
19 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.   DOI   ScienceOn
20 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.   DOI
21 Erlich S, Mizrachy L, Segev O, et al (2007). Differential interactions between Beclin 1 and Bcl-2 family members. Autophagy, 3, 561-8.   DOI
22 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.   DOI   ScienceOn
23 Carew JS, Nawrocki ST, Cleveland JL (2007). Modulating autophagy for therapeutic benefit. Autophagy, 3, 464-7.   DOI
24 Chen N, Karantza-Wadsworth V (2009). Role and regulation of autophagy in cancer. Biochim Biophys Acta, 1793, 1516-23.   DOI   ScienceOn
25 Gewirtz DA (2007). Autophagy as a mechanism of radiation sensitization in breast tumor cells. Autophagy, 3, 249-50.   DOI
26 Gozuacik D, Kimchi A (2004). Autophagy as a cell death and tumor suppressor mechanism. Oncogene, 23, 2891-906.   DOI   ScienceOn
27 Hanahan D, Weinberg RA (2000). The hallmarks of cancer. Cell, 100, 57-70.   DOI   ScienceOn
28 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.   DOI   ScienceOn
29 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.
30 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.   DOI   ScienceOn
31 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.   DOI   ScienceOn
32 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.   DOI   ScienceOn