Suppression of autophagic genes sensitizes CUG2-overexpressing A549 human lung cancer cells to oncolytic vesicular stomatitis virus-induced apoptosis.
We showed in our previous study that cancer upregulated gene (CUG) 2, a novel oncogene, confers resistance to infection of oncolytic vesicular stomatitis virus (VSV) by activating Stat1-mediated signal transduction. Since many studies have reported that autophagy is involved in virus replication, we investigated whether autophagy also plays a role in the antiviral activity in A549 cells overexpressing CUG2 (A549-CUG2). We suppressed Atg5 or Beclin 1 expression using siRNA and examined its effect on the susceptibility of cells to infection by oncolytic VSV. We found that A549-CUG2 cells treated with Atg5 or Beclin 1 siRNA became susceptible to VSV infection, whereas A549-CUG2 cells treated with control siRNA were resistant. This result suggests that autophagy is involved in the antiviral response of A549-CUG2 cells. Further investigation revealed that autophagy impairment enhanced the generation of reactive oxygen species (ROS), which resulted in inactivation of S6 kinase. Under these conditions, the levels of ISG15 transcript and protein decreased, which conferred on A549-CUG2 cell susceptibility to VSV infection. Finally, we found that overloading of H₂O₂ sensitized control A549-CUG2 cells to VSV-induced apoptosis. Taken together, these results indicate that autophagy impairment induces excessive ROS formation, which decreases S6 kinase activity and ISG15 expression, ultimately rendering the A549-CUG2 cells susceptible to VSV infection. We propose that autophagy impairment is a potential strategy for successful VSV virotherapy of CUG2-overexpressing tumors.
BK21+, Department of Cogno-Mechatronics Engineering, Pusan National University, Busan 609-735, Republic of Korea.
Waraporn
W
Malilas
Immunotherapy Research Center, Korea Research Institute of Bioscience and Biotechnology, and Department of Functional Genomics, University of Science and Technology, Daejeon 305-333, Republic of Korea.
Sang Seok
SS
Koh
Department of Microbiology and Molecular Biology, Chungnam National University, Daejeon 305-764, Republic of Korea.
Soojin
S
Lee
BK21+, Department of Cogno-Mechatronics Engineering, Pusan National University, Busan 609-735, Republic of Korea.
Ratakorn
R
Srisuttee
BK21+, Department of Cogno-Mechatronics Engineering, Pusan National University, Busan 609-735, Republic of Korea.
Il-Rae
IR
Cho
BK21+, Department of Cogno-Mechatronics Engineering, Pusan National University, Busan 609-735, Republic of Korea.
Jeong
J
Moon
BK21+, Department of Cogno-Mechatronics Engineering, Pusan National University, Busan 609-735, Republic of Korea.
Sirichat
S
Kaowinn
Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, T2N4N1 Alberta, Canada.
Randal N
RN
Johnston
BK21+, Department of Cogno-Mechatronics Engineering, Pusan National University, Busan 609-735, Republic of Korea.
Young-Hwa
YH
Chung
D000818:Animals; D017209:Apoptosis / Q000235:genetics*; D051017:Apoptosis Regulatory Proteins / Q000235:genetics; D001343:Autophagy / Q000235:genetics*; D000071187:Autophagy-Related Protein 5; D000071186:Beclin-1; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000096:biosynthesis / Q000235:genetics*; D016207:Cytokines / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008175:Lung Neoplasms / Q000821:virology; D008565:Membrane Proteins / Q000235:genetics; D051379:Mice; D008869:Microtubule-Associated Proteins / Q000235:genetics; D050504:Oncolytic Viruses / Q000235:genetics / Q000472:pathogenicity; D034622:RNA Interference; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species / Q000378:metabolism; D018353:Rhabdoviridae Infections / Q000235:genetics / Q000473:pathology; D019893:Ribosomal Protein S6 Kinases / Q000037:antagonists & inhibitors; D050794:STAT1 Transcription Factor / Q000235:genetics; D015398:Signal Transduction; D014452:Ubiquitins / Q000096:biosynthesis / Q000235:genetics*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics / Q000472:pathogenicity*; D014779:Virus Replication
C502778:ATG5 protein, human; D051017:Apoptosis Regulatory Proteins; D000071187:Autophagy-Related Protein 5; C491997:BECN1 protein, human; D000071186:Beclin-1; C534000:CENPW protein, human; D002868:Chromosomal Proteins, Non-Histone; D016207:Cytokines; D008565:Membrane Proteins; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; D017382:Reactive Oxygen Species; D050794:STAT1 Transcription Factor; C494079:STAT1 protein, human; D014452:Ubiquitins; C074034:ISG15 protein, human; D019893:Ribosomal Protein S6 Kinases