Autophagy is involved in recombinant Newcastle disease virus (rL-RVG)-induced cell death of stomach adenocarcinoma cells in vitro

Bu,Xu-Feng; Wang,Mu-Bing; Zhang,Zhi-Jian; Zhao,Ying-Hai; Li,Mi; Yan,Yu-Lan

doi:10.3892/ijo.2015.3039

Autophagy is involved in recombinant Newcastle disease virus (rL-RVG)-induced cell death of stomach adenocarcinoma cells in vitro

Authors:
- Xu-Feng Bu
- Mu-Bing Wang
- Zhi-Jian Zhang
- Ying-Hai Zhao
- Mi Li
- Yu-Lan Yan
View Affiliations

Affiliations: Department of General Surgery, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, P.R. China, Clinical Medicine College of Jiangsu University, Zhenjiang, Jiangsu, P.R. China, Department of Internal Medicine, Affiliated People's Hospital of Jiangsu University, Zhenjiang, Jiangsu, P.R. China
Published online on: June 9, 2015 https://doi.org/10.3892/ijo.2015.3039
Pages: 679-689

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Oncolytic viruses can kill malignant cells while sparing normal cells. Multiple pathways are involved in this action. The antitumor effects of viral infection on SGC-7901 and AGS cells were investigated. We measured endoplasmic reticulum stress and autophagy caused by the recombinant avirulent Newcastle disease virus (NDV) LaSota strain expressing the rabies virus glycoprotein (rL-RVG) and the NDV wild-type strain. The dose-response curves were analyzed using the MTT assay. The expression of RVG was detected by western blotting, RT-PCR and immunofluorescence analyses. Cell death and autophagy were observed using transmission electron microscopy, TUNEL and western blotting. Endoplasmic reticulum stress and the mitochondrial transmembrane potential were detected by western blotting and immunoﬂuorescence, respectively. Immunoﬂuorescence, western blot and RT-PCR analyses indicated that RVG gene and protein were expressed in SGC-7901 and AGS cells infected by rL-RVG. MTT and TUNEL analyses showed that the growth of SGC-7901 and AGS cells in the rL-RVG-infected group was significantly inhibited compared with the wild-type NDV-infected group (p<0.05). Western blot analysis indicated that rL-RVG and NDV induced increases in apoptosis, endoplasmic reticulum stress, and autophagy in the SGC-7901 and AGS cells. However, apoptosis and autophagy decreased in these cells after the application of the autophagy pathway inhibitor 3-MA or ATG-5-specific siRNA. Immunoﬂuorescence analysis showed that the mitochondrial membrane potential collapsed. Taken together, these results indicate that the rL-RVG virus group is much more powerful compared with the NDV-infected group (p<0.05). rL-RVG and NDV are potent antitumor agents that induce autophagy.

View Figures

View References

1	Chen J, Shen W, Xia J, Xu R, Zhu M and Xu M: Effect of S-1 maintenance chemotherapy following DCF regimen in patients with advanced gastric cancer. Nan Fang Yi Ke Da Xue Xue Bao. 34:1057–1060. 2014.(In Chinese). PubMed/NCBI
2	Khalighinejad N, Hariri H, Behnamfar O, Yousefi A and Momeni A: Adenoviral gene therapy in gastric cancer: A review. World J Gastroenterol. 14:180–184. 2008. View Article : Google Scholar : PubMed/NCBI
3	Janke M, Peeters B, de Leeuw O, Moorman R, Arnold A, Fournier P and Schirrmacher V: Recombinant Newcastle disease virus (NDV) with inserted gene coding for GM-CSF as a new vector for cancer immunogene therapy. Gene Ther. 14:1639–1649. 2007. View Article : Google Scholar : PubMed/NCBI
4	Beutner U, Lorenz U, Illert B, Rott L, Timmermann W, Vollmers HP, Müller-Hermelink HK, Thiede A and Ulrichs K: Neoadjuvant therapy of gastric cancer with the human monoclonal IgM antibody SC-1: Impact on the immune system. Oncol Rep. 19:761–769. 2008.PubMed/NCBI
5	Garg AD and Agostinis P: ER stress, autophagy and immunogenic cell death in photodynamic therapy-induced anti-cancer immune responses. Photochem Photobiol Sci. 13:474–487. 2014. View Article : Google Scholar : PubMed/NCBI
6	Hossain A, Radwan FF, Doonan BP, God JM, Zhang L, Bell PD and Haque A: A possible cross-talk between autophagy and apoptosis in generating an immune response in melanoma. Apoptosis. 17:1066–1078. 2012. View Article : Google Scholar : PubMed/NCBI
7	Puhlmann J, Puehler F, Mumberg D, Boukamp P and Beier R: Rac1 is required for oncolytic NDV replication in human cancer cells and establishes a link between tumorigenesis and sensitivity to oncolytic virus. Oncogene. 29:2205–2216. 2010. View Article : Google Scholar : PubMed/NCBI
8	Pecora AL, Rizvi N, Cohen GI, Meropol NJ, Sterman D, Marshall JL, Goldberg S, Gross P, O’Neil JD, Groene WS, et al: Phase I trial of intravenous administration of PV701, an oncolytic virus, in patients with advanced solid cancers. J Clin Oncol. 20:2251–2266. 2002. View Article : Google Scholar : PubMed/NCBI
9	Sinkovics JG and Horvath JC: Newcastle disease virus (NDV): Brief history of its oncolytic strains. J Clin Virol. 16:1–15. 2000. View Article : Google Scholar : PubMed/NCBI
10	Yang S, Liu W, Cui H, Sun S and Wang J: In vitro induction of apoptosis in tumor cells by inactivated NDV and IAV. Cancer Biother Radiopharm. 22:200–205. 2007. View Article : Google Scholar : PubMed/NCBI
11	Yaacov B, Eliahoo E, Lazar I, Ben-Shlomo M, Greenbaum I, Panet A and Zakay-Rones Z: Selective oncolytic effect of an attenuated Newcastle disease virus (NDV-HUJ) in lung tumors. Cancer Gene Ther. 15:795–807. 2008. View Article : Google Scholar : PubMed/NCBI
12	Lam HY, Yeap SK, Rasoli M, Omar AR, Yusoff K, Suraini AA and Alitheen NB: Safety and clinical usage of newcastle disease virus in cancer therapy. J Biomed Biotechnol. 2011:7187102011. View Article : Google Scholar : PubMed/NCBI
13	Lay S, Préhaud C, Dietzschold B and Lafon M: Glycoprotein of nonpathogenic rabies viruses is a major inducer of apoptosis in human jurkat T cells. Ann NY Acad Sci. 1010:577–581. 2003. View Article : Google Scholar
14	Préhaud C, Lay S, Dietzschold B and Lafon M: Glycoprotein of nonpathogenic rabies viruses is a key determinant of human cell apoptosis. J Virol. 77:10537–10547. 2003. View Article : Google Scholar : PubMed/NCBI
15	Vähä-Koskela MJ, Heikkilä JE and Hinkkanen AE: Oncolytic viruses in cancer therapy. Cancer Lett. 254:178–216. 2007. View Article : Google Scholar : PubMed/NCBI
16	Klionsky DJ and Emr SD: Autophagy as a regulated pathway of cellular degradation. Science. 290:1717–1721. 2000. View Article : Google Scholar : PubMed/NCBI
17	Rubinsztein DC, Codogno P and Levine B: Autophagy modulation as a potential therapeutic target for diverse diseases. Nat Rev Drug Discov. 11:709–730. 2012. View Article : Google Scholar : PubMed/NCBI
18	Carroll RG and Martin SJ: Autophagy in multiple myeloma: What makes you stronger can also kill you. Cancer Cell. 23:425–426. 2013. View Article : Google Scholar : PubMed/NCBI
19	Kroemer G and Levine B: Autophagic cell death: The story of a misnomer. Nat Rev Mol Cell Biol. 9:1004–1010. 2008. View Article : Google Scholar : PubMed/NCBI
20	Ahn JH and Lee M: Autophagy-dependent survival of mutant B-Raf melanoma cells selected for resistance to apoptosis induced by inhibitors against oncogenic B-Raf. Biomol Ther (Seoul). 21:114–120. 2013. View Article : Google Scholar
21	Martín V, Sanchez-Sanchez AM, Puente-Moncada N, Gomez-Lobo M, Alvarez-Vega MA, Antolín I and Rodriguez C: Involvement of autophagy in melatonin-induced cytotoxicity in glioma-initiating cells. J Pineal Res. 57:308–316. 2014. View Article : Google Scholar : PubMed/NCBI
22	Xu ZX, Liang J, Haridas V, Gaikwad A, Connolly FP, Mills GB and Gutterman JU: A plant triterpenoid, avicin D, induces autophagy by activation of AMP-activated protein kinase. Cell Death Differ. 14:1948–1957. 2007. View Article : Google Scholar : PubMed/NCBI
23	Cheng J, Wei HL, Chen J and Xie B: Antitumor effect of arsenic trioxide in human K562 and K562/ADM cells by autophagy. Toxicol Mech Methods. 22:512–519. 2012.PubMed/NCBI
24	Kulich I and Žárský V: Autophagy-related direct membrane import from ER/cytoplasm into the vacuole or apoplast: A hidden gateway also for secondary metabolites and phytohormones? Int J Mol Sci. 15:7462–7474. 2014. View Article : Google Scholar : PubMed/NCBI
25	Shailasree S, Venkataramana M, Niranjana SR and Prakash HS: Cytotoxic effect of p-coumaric acid on neuroblastoma, N2a cell via generation of reactive oxygen species leading to dysfunction of mitochondria inducing apoptosis and autophagy. Mol Neurobiol. 51:119–130. 2015. View Article : Google Scholar
26	Ge J, Wang X, Tao L, Wen Z, Feng N, Yang S, Xia X, Yang C, Chen H and Bu Z: Newcastle disease virus-vectored rabies vaccine is safe, highly immunogenic, and provides long-lasting protection in dogs and cats. J Virol. 85:8241–8252. 2011. View Article : Google Scholar : PubMed/NCBI
27	Yan Y, Liang B, Zhang J, Liu Y and Bu X: Apoptotic induction of lung adenocarcinoma A549 cells infected by recombinant RVG Newcastle disease virus (rLRVG) in vitro. Mol Med Rep. 11:317–326. 2015.
28	Chen W, Sun Y, Liu K and Sun X: Autophagy: A double-edged sword for neuronal survival after cerebral ischemia. Neural Regen Res. 9:1210–1216. 2014. View Article : Google Scholar : PubMed/NCBI
29	Gong A, Ye S, Xiong E, Guo W, Zhang Y, Peng W, Shao G, Jin J, Zhang Z, Yang J, et al: Autophagy contributes to ING4-induced glioma cell death. Exp Cell Res. 319:1714–1723. 2013. View Article : Google Scholar : PubMed/NCBI
30	Yousefi S, Perozzo R, Schmid I, Ziemiecki A, Schaffner T, Scapozza L, Brunner T and Simon HU: Calpain-mediated cleavage of Atg5 switches autophagy to apoptosis. Nat Cell Biol. 8:1124–1132. 2006. View Article : Google Scholar : PubMed/NCBI
31	Yuzefovych LV, LeDoux SP, Wilson GL and Rachek LI: Mitochondrial DNA damage via augmented oxidative stress regulates endoplasmic reticulum stress and autophagy: Crosstalk, links and signaling. PLoS One. 8:e833492013. View Article : Google Scholar : PubMed/NCBI
32	Zorn U, Dallmann I, Grosse J, Kirchner H, Poliwoda H and Atzpodien J: Induction of cytokines and cytotoxicity against tumor cells by Newcastle disease virus. Cancer Biother. 9:225–235. 1994. View Article : Google Scholar : PubMed/NCBI
33	Tsuchihara K, Fujii S and Esumi H: Autophagy and cancer: Dynamism of the metabolism of tumor cells and tissues. Cancer Lett. 278:130–138. 2009. View Article : Google Scholar
34	Kimura T, Takabatake Y, Takahashi A and Isaka Y: Chloroquine in cancer therapy: A double-edged sword of autophagy. Cancer Res. 73:3–7. 2013. View Article : Google Scholar : PubMed/NCBI
35	Mariño G, Martins I and Kroemer G: Autophagy in Ras-induced malignant transformation: Fatal or vital? Mol Cell. 42:1–3. 2011. View Article : Google Scholar : PubMed/NCBI
36	Gong JS and Kim GJ: The role of autophagy in the placenta as a regulator of cell death. Clin Exp Reprod Med. 41:97–107. 2014. View Article : Google Scholar : PubMed/NCBI
37	Luo S and Rubinsztein DC: BCL2L11/BIM: A novel molecular link between autophagy and apoptosis. Autophagy. 9:104–105. 2013. View Article : Google Scholar :
38	Saeki K, Yuo A, Okuma E, Yazaki Y, Susin SA, Kroemer G and Takaku F: Bcl-2 down-regulation causes autophagy in a caspase-independent manner in human leukemic HL60 cells. Cell Death Differ. 7:1263–1269. 2000. View Article : Google Scholar
39	Lomonosova E and Chinnadurai G: BH3-only proteins in apoptosis and beyond: An overview. Oncogene. 27(Suppl 1): S2–S19. 2008. View Article : Google Scholar
40	Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N, Packer M, Schneider MD and Levine B: Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell. 122:927–939. 2005. View Article : Google Scholar : PubMed/NCBI
41	Cho DH, Jo YK, Hwang JJ, Lee YM, Roh SA and Kim JC: Caspase-mediated cleavage of ATG6/Beclin-1 links apoptosis to autophagy in HeLa cells. Cancer Lett. 274:95–100. 2009. View Article : Google Scholar
42	Lin CJ, Lee CC, Shih YL, Lin CH, Wang SH, Chen TH and Shih CM: Inhibition of mitochondria- and endoplasmic reticulum stress-mediated autophagy augments temozolomide-induced apoptosis in glioma cells. PLoS One. 7:e387062012. View Article : Google Scholar : PubMed/NCBI
43	Jheng JR, Ho JY and Horng JT: ER stress, autophagy, and RNA viruses. Front Microbiol. 5:3882014. View Article : Google Scholar : PubMed/NCBI
44	Digaleh H, Kiaei M and Khodagholi F: Nrf2 and Nrf1 signaling and ER stress crosstalk: Implication for proteasomal degradation and autophagy. Cell Mol Life Sci. 70:4681–4694. 2013. View Article : Google Scholar : PubMed/NCBI
45	Tian J, Hu X and Qu Q: Effect and mechanism of endoplasmic reticulum stress on cisplatin resistance in ovarian carcinoma]. Zhonghua Zhong Liu Za Zhi. 36:324–328. 2014.(In Chinese). PubMed/NCBI
46	Suzuki SW, Onodera J and Ohsumi Y: Starvation induced cell death in autophagy-defective yeast mutants is caused by mitochondria dysfunction. PLoS One. 6:e174122011. View Article : Google Scholar : PubMed/NCBI
47	Meng C, Zhou Z, Jiang K, Yu S, Jia L, Wu Y, Liu Y, Meng S and Ding C: Newcastle disease virus triggers autophagy in U251 glioma cells to enhance virus replication. Arch Virol. 157:1011–1018. 2012. View Article : Google Scholar : PubMed/NCBI
48	Meng G, Xia M, Wang D, Chen A, Wang Y, Wang H, Yu D and Wei J: Mitophagy promotes replication of oncolytic Newcastle disease virus by blocking intrinsic apoptosis in lung cancer cells. Oncotarget. 5:6365–6374. 2014.PubMed/NCBI
49	Marzetti E, Csiszar A, Dutta D, Balagopal G, Calvani R and Leeuwenburgh C: Role of mitochondrial dysfunction and altered autophagy in cardiovascular aging and disease: From mechanisms to therapeutics. Am J Physiol Heart Circ Physiol. 305:H459–H476. 2013. View Article : Google Scholar : PubMed/NCBI
50	Wu JJ, Quijano C, Chen E, Liu H, Cao L, Fergusson MM, Rovira II, Gutkind S, Daniels MP, Komatsu M, et al: Mitochondrial dysfunction and oxidative stress mediate the physiological impairment induced by the disruption of autophagy. Aging (Albany, NY). 1:425–437. 2009.
51	Lee J, Giordano S and Zhang J: Autophagy, mitochondria and oxidative stress: Cross-talk and redox signalling. Biochem J. 441:523–540. 2012. View Article : Google Scholar :