Apogossypolone induces apoptosis and autophagy in nasopharyngeal carcinoma cells in an in vitro and in vivo study

Zheng,Ruinian; Chen,Kexu; Zhang,Yu; Huang,Jie; Shi,Fengrong; Wu,Gang; Wang,Senming

doi:10.3892/ol.2017.6176

Apogossypolone induces apoptosis and autophagy in nasopharyngeal carcinoma cells in an in vitro and in vivo study

Authors:
- Ruinian Zheng
- Kexu Chen
- Yu Zhang
- Jie Huang
- Fengrong Shi
- Gang Wu
- Senming Wang
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Affiliations: Department of Oncology, Dongguan People's Hospital, Dongguan, Guangdong 523000, P.R. China, Department of Oncology, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510282, P.R. China
Published online on: May 16, 2017 https://doi.org/10.3892/ol.2017.6176
Pages: 751-757
Copyright: © Zheng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Nasopharyngeal carcinoma (NPC) has a high incidence and mortality rate, particularly in Southern China. Apogossypolone (ApoG2) is a novel derivative of gossypol with antitumor activity and less toxicity. The human NPC CNE‑2 cell line was studied in the in vitro model; whilst 4 week‑old male nude mice (BALB/c‑nu) were inoculated subcutaneously with CNE‑2 cells, and xenograft tumors were studied in the in vivo model. Graded concentrations of ApoG2 were used in treatment studies. In ApoG2‑treated and control in vitro and in vivo tumor cells, cell apoptosis, and autophagy were evaluated and quantified using fluorescent and transmission electron microscopy and flow cytometry. Hoechst‑33258 fluorescence staining was used to evaluate apoptosis in treated and non‑treated cell culture and xenograft NPC cells. Western blotting was performed on lysed tumor cells using primary antibodies to B‑cell lymphoma‑2 (Bcl‑2), beclin‑1, and β‑actin, and flow cytometry results indicated cell apoptosis rates of 3.90±0.34 and 19.52±1.18% in the control and ApoG2‑treated cells, respectively (F=485.294, P<0.001). Western blot analysis showed that ApoG2 significantly decreased expression of the Bcl‑2 protein in CNE‑2 cells, when compared with control cells (F=68.909, P=0.001) and flow cytometry showed cell autophagy rates of 0.92±3.10% of control cells compared with 28.24±7.35% of ApoG2‑treated cells (F=31.035, P=0.003). ApoG2 treatment significantly increased beclin‑1 protein expression in CNE‑2 cells (F=497.906, P<0.001). ApoG2 treatment inhibited NPC xenograft tumor growth by 65.49% (P<0.05). In conclusion, these results support a role for ApoG2 in inhibiting the growth of human NPC cells by inducing apoptosis and autophagy. Future controlled clinical studies could be planned, to define safety, efficacy and dosing regimens for ApoG2 as a potential treatment for patients with NPC.

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1	Wang Y, Zhang Y and Ma S: Racial differences in nasopharyngeal carcinoma in the United States. Cancer Epidemiol. 37:793–802. 2013. View Article : Google Scholar : PubMed/NCBI
2	Kimura Y, Suzuki D, Tokunaga T, Takabayashi T, Yamada T, Wakisaka N, Yoshizaki T, Murata H, Miwa K, Shoujaku H, et al: Epidemiological analysis of nasopharyngeal carcinoma in the central region of Japan during the period from 1996 to 2005. Auris Nasus Larynx. 38:244–249. 2011. View Article : Google Scholar : PubMed/NCBI
3	Lau HY, Leung CM, Chan YH, Lee AW, Kwong DL, Lung ML and Lam TH: Secular trends of salted fish consumption and nasopharyngeal carcinoma: A multi-jurisdiction ecological study in 8 regions from 3 continents. BMC Cancer. 13:2982013. View Article : Google Scholar : PubMed/NCBI
4	Xu T, Shen C, Ou X, He X, Ying H and Hu C: The role of adjuvant chemotherapy in nasopharyngeal carcinoma with bulky neck lymph nodes in the era of IMRT. Oncotarget. 7:21013–21022. 2016.PubMed/NCBI
5	Xu T, Liu Y, Dou S, Li F, Guan X and Zhu G: Weekly cetuximab concurrent with IMRT aggravated radiation-induced oral mucositis in locally advanced nasopharyngeal carcinoma: Results of a randomized phase II study. 51:875–879. 2015.
6	Li H, Wang DL, Liu XW, Chen MY, Mo YX, Geng ZJ and Xie CM: MRI signal changes in the skull base bone after endoscopic nasopharyngectomy for recurrent NPC: A serial study of 9 patients. Eur J Radiol. 82:309–315. 2013. View Article : Google Scholar : PubMed/NCBI
7	Zhan YH, Huang XF, Hu XB, An QX, Liu ZX and Zhang XQ: Growth inhibition and apoptosis induction of human umbilical vein endothelial cells by apogossypolone. Asian Pac J Cancer Prev. 14:1791–1795. 2013. View Article : Google Scholar : PubMed/NCBI
8	Niu X, Li S, Wei F, Huang J, Wu G, Xu L, Xu D and Wang S: Apogossypolone induces autophagy and apoptosis in breast cancer MCF-7 cells in vitro and in vivo. Breast Cancer. 21:223–230. 2014. View Article : Google Scholar : PubMed/NCBI
9	Xin J, Zhan YH, Xia LM, Zhu HW, Nie YZ, Liang JM and Tian J: ApoG2 as the most potent gossypol derivatives inhibits cell growth and induces apoptosis on gastric cancer cells. Biomed Pharmacother. 67:88–95. 2013. View Article : Google Scholar : PubMed/NCBI
10	Banerjee S, Choi M, Aboukameel A, Wang Z, Mohammad M, Chen J, Yang D, Sarkar FH and Mohammad RM: Preclinical studies of apogossypolone, a novel pan inhibitor of bcl-2 and mcl-1, synergistically potentiates cytotoxic effect of gemcitabine in pancreatic cancer cells. Pancreas. 39:323–331. 2010. View Article : Google Scholar : PubMed/NCBI
11	Lin J, Wu YJ, Yang DJ and Zhao YQ: Effect of apogossypolone on induction apoptosis in multiple myeloma cells and its mechanisms. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 17:92–98. 2009.(In Chinese). PubMed/NCBI
12	Balakrishnan K, Aggarwal S, Wierda W and Gandhi V: Bax and Bak are required for apogossypolone, a BH3-mimetic, induced apoptosis in chronic lymphocytic leukemia cells. Leuk Lymphoma. 54:1097–1100. 2013. View Article : Google Scholar : PubMed/NCBI
13	Cheng P, Ni Z, Dai X, Wang B, Ding W, Rae Smith A, Xu L, Wu D, He F and Lian J: The novel BH-3 mimetic apogossypolone induces Beclin-1- and ROS-mediated autophagy in human hepatocellular carcinoma [corrected] cells. Cell Death Dis. 4:e4892013. View Article : Google Scholar : PubMed/NCBI
14	Sun J, Li ZM, Hu ZY, Lin XB, Zhou NN, Xian LJ, Yang DJ and Jiang WQ: ApoG2 inhibits antiapoptotic Bcl-2 family proteins and induces mitochondria-dependent apoptosis in human lymphoma U937 cells. Anticancer Drugs. 19:967–974. 2008. View Article : Google Scholar : PubMed/NCBI
15	Hockenbery DM: Bcl-2 in cancer, development and apoptosis. J Cell Sci Suppl. 18:51–55. 1994. View Article : Google Scholar : PubMed/NCBI
16	Raffo AJ, Perlman H, Chen MW, Day ML, Streitman JS and Buttyan R: Overexpression of bcl-2 protects prostate cancer cells from apoptosis in vitro and confers resistance to androgen depletion in vivo. Cancer Res. 55:4438–4445. 1995.PubMed/NCBI
17	National Research Council (US), . Committee for the Update of the Guide for the Care and Use of Laboratory Animals. ‘Guide for the Care and Use of Laboratory Animals. ’ Guide for the Care & Use of Laboratory Animals. 103:1072–1073. 2011.
18	Xu ZJ, Zheng RS, Zhang SW, Zou XN and Chen WQ: Nasopharyngeal carcinoma incidence and mortality in China in 2009. Chin J Cancer. 32:453–460. 2013. View Article : Google Scholar : PubMed/NCBI
19	Mesia R, Pastor M, Grau JJ and del Barco E; SEOM: SEOM clinical guidelines for the treatment of nasopharyngeal carcinoma 2013. Clin Transl Oncol. 15:1025–1029. 2013. View Article : Google Scholar : PubMed/NCBI
20	Piro LD: Apoptosis, Bcl-2 antisense, and cancer therapy. Oncology (Williston Park). 18 13 Suppl 10:S5–S10. 2004.
21	Kontos CK, Christodoulou MI and Scorilas A: Apoptosis-related BCL2-family members: Key players in chemotherapy. Anticancer Agents Med Chem. 14:353–374. 2014. View Article : Google Scholar : PubMed/NCBI
22	Kaneko T, Zhang Z, Mantellini MG, Karl E, Zeitlin B, Verhaegen M, Soengas MS, Lingen M, Strieter RM, Nunez G and Nör JE: Bcl-2 orchestrates a cross-talk between endothelial and tumor cells that promotes tumor growth. Cancer Res. 67:9685–9693. 2007. View Article : Google Scholar : PubMed/NCBI
23	Tucker CA, Kapanen AI, Chikh G, Hoffman BG, Kyle AH, Wilson IM, Masin D, Gascoyne RD, Bally M and Klasa RJ: Silencing Bcl-2 in models of mantle cell lymphoma is associated with decreases in cyclin D1, nuclear factor-kappaB, p53, bax, and p27 levels. Mol Cancer Ther. 7:749–758. 2008. View Article : Google Scholar : PubMed/NCBI
24	Tekedereli I, Alpay SN, Akar U, Yuka E, Ayugo-Rodriguez C, Han HD, Sood AK, Lopez-Berestein G and Ozpolat B: Therapeutic silencing of Bcl-2 by systemically administered siRNA nanotherapeutics inhibits tumor growth by autophagy and apoptosis and enhances the efficacy of chemotherapy in orthotopic xenograft models of ER (−) and ER (+) breast cancer. Mol Ther Nucleic Acids. 2:e1212013. View Article : Google Scholar : PubMed/NCBI
25	Du P, Cao H, Wu HR, Zhu BS, Wang HW, Gu CW, Xing CG and Chen W: Blocking Bcl-2 leads to autophagy activation and cell death of the HEPG2 liver cancer cell line. Asian Pac J Cancer Prev. 14:5849–5854. 2013. View Article : Google Scholar : PubMed/NCBI
26	Akar U, Chaves-Reyez A, Barria M, Tari A, Sanguino A, Kondo Y, Kondo S, Arun B, Lopez-Berestein G and Ozpolat B: Silencing of Bcl-2 expression by small interfering RNA induces autophagic cell death in MCF-7 breast cancer cells. Autophagy. 4:669–679. 2008. View Article : Google Scholar : PubMed/NCBI
27	Paoluzzi L, Gonen M, Gardner JR, Mastrella J, Yang D, Holmlund J, Sorenen M, Leopold L, Manova K, Marcucci G, et al: Targeting Bcl-2 family members with the BH3 mimetic AT-101 markedly enhances the therapeutic effects of chemotherapeutic agents in in vitro and in vivo models of B-cell lymphoma. Blood. 111:5350–5358. 2008. View Article : Google Scholar : PubMed/NCBI
28	Lieber J, Kirchner B, Eicher C, Warman SW, Seitz G, Fuchs J and Armeanu-Ebinger S: Inhibition of Bcl-2 and Bcl-X enhances chemotherapy sensitivity in hepatoblastoma cells. Pediatr Blood Cancer. 55:1089–1095. 2010. View Article : Google Scholar : PubMed/NCBI
29	Lamb CA, Yoshimori T and Tooze SA: The autophagosome: Origins unknown, biogenesis complex. Nat Rev Mol Cell Biol. 14:759–774. 2013. View Article : Google Scholar : PubMed/NCBI
30	Yang Z and Klionsky DJ: Eaten alive: A history of macroautophagy. Nat Cell Biol. 12:814–822. 2010. View Article : Google Scholar : PubMed/NCBI
31	Xu HD, Wu D, Gu JH, Ge JB, Wu JC, Han R, Liang ZQ and Qin ZH: The pro-survival role of autophagy depends on Bcl-2 under nutrition stress conditions. PLoS One. 8:e632322013. View Article : Google Scholar : PubMed/NCBI
32	Tian S, Lin J, Jun Zhou J, Wang X, Li Y, Ren X, Yu W, Zhong W, Xiao J, Sheng F, et al: Beclin 1-independent autophagy induced by a Bcl-XL/Bcl-2 targeting compound, Z18. Autophagy. 6:1032–1041. 2010. View Article : Google Scholar : PubMed/NCBI
33	He JH, Liao XL, Wang W, Li DD, Chen WD, Deng R, Yang D, Han ZP, Jiang JW and Zhu XF: Apogossypolone, a small-molecule inhibitor of Bcl-2, induces radiosensitization of nasopharyngeal carcinoma cells by stimulating autophagy. Int J Oncol. 45:1099–1108. 2014.PubMed/NCBI
34	Hu ZY, Wang J, Cheng G, Zhu XF, Huang P, Yang D and Zeng YX: Apogossypolone targets mitochondria and light enhances its anticancer activity by stimulating generation of singlet oxygen and reactive oxygen species. Chin J Cancer. 30:41–53. 2011. View Article : Google Scholar : PubMed/NCBI
35	Hu ZY, Sun J, Zhu XF, Yang D and Zeng YX: ApoG2 induces cell cycle arrest of nasopharyngeal carcinoma cells by suppressing the c-Myc signaling pathway. J Transl Med. 7:742009. View Article : Google Scholar : PubMed/NCBI
36	Hu ZY, Zhu XF, Zhong ZD, Sun J, Wang J, Yang D and Zeng YX: ApoG2, a novel inhibitor of antiapoptotic Bcl-2 family proteins, induces apoptosis and suppresses tumor growth in nasopharyngeal carcinoma xenografts. Int J Cancer. 123:2418–2429. 2008. View Article : Google Scholar : PubMed/NCBI