caa a22 4500 445795808 CHVBK 20180317145124.0 cr unu---uuuuu 170323e20110401xx s 000 0 eng 10.1007/s10620-010-1420-1 doi (NATIONALLICENCE)springer-10.1007/s10620-010-1420-1 Helicobacter pylori VacA Reduces the Cellular Expression of STAT3 and Pro-survival Bcl-2 Family Proteins, Bcl-2 and Bcl-XL, Leading to Apoptosis in Gastric Epithelial Cells [Elektronische Daten] [Ayako Matsumoto, Hajime Isomoto, Masaaki Nakayama, Junzo Hisatsune, Yoshito Nishi, Yujiro Nakashima, Kayoko Matsushima, Hisao Kurazono, Kazuhiko Nakao, Toshiya Hirayama, Shigeru Kohno] Background: Helicobacter pylori vacuolating cytotoxin, VacA, stimulates apoptosis via a mitochondria-dependent pathway. VacA induces apoptosis via activation of the pro-apoptotic B-cell lymphoma (Bcl)-2 family proteins, Bcl-2-associated X protein (Bax) and Bcl-2 homologous antagonist/killer (Bak), while the implication of such pro-survival Bcl-2 family members as Bcl-2 and Bcl-XL in the VacA-induced apoptosis remains unknown. Signal transduction and activator of transcription 3 (STAT3) is a pivotal transcription factor that upregulates Bcl-2 and Bcl-XL. Aims: This study was conducted to elicit the implication of STAT3 and pro-survival Bcl-2 and Bcl-XL in the intrinsic apoptosis. Methods: Immunoblot and reverse transcriptase real-time polymerase chain reaction (RT-PCR) were employed to assess the cellular expression of STAT3, Bcl-2, and Bcl-XL in response to purified VacA in gastric adenocarcinoma cell lines. VacA-induced apoptosis was quantitated morphologically following knockdown by each specific small interfering RNA (siRNA) or in the presence of pharmacological inhibitors. Results: VacA reduced STAT3, Bcl-2, and Bcl-XL expression in a dose-dependent manner. Knockdown of STAT3, Bcl-2, and Bcl-XL by siRNA induced apoptosis to a similar extent in the case of sufficient VacA inoculation. The VacA-mediated reduction of STAT3 expression was independent of cellular vacuolization, since a vacuolar-type ATPase inhibitor, bafilomycin A1, did not inhibit VacA-induced reduction of STAT3, Bcl-2, and Bcl-XL expression. Instead, a c-JUN NH2-terminal kinase (JNK) inhibitor, SP600125, restored the VacA-induced reduction of STAT3 expression to the basal level. Conclusions: VacA-induced apoptosis may be, in part, implicated in the reduction of STAT3 linking to the downregulation of Bcl-2 and Bcl-XL, in association with JNK activity. Springer Science+Business Media, LLC, 2010 Vacuolating cytotoxin, VacA nationallicence Signal transduction and activator of transcription 3, STAT3 nationallicence B-cell lymphoma (Bcl)-2 and Bcl-XL nationallicence c-JUN NH2-terminal kinase, JNK nationallicence Apoptosis nationallicence Matsumoto Ayako Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki University School of Medicine, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut Isomoto Hajime Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut Nakayama Masaaki Department of Bacteriology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, 852-8523, Nagasaki, Japan aut Hisatsune Junzo Department of Bacteriology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, 852-8523, Nagasaki, Japan aut Nishi Yoshito Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki University School of Medicine, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut Nakashima Yujiro Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut Matsushima Kayoko Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut Kurazono Hisao Department of Applied Veterinary Medicine and Public Health, Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Obihiro, Japan aut Nakao Kazuhiko Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut Hirayama Toshiya Department of Bacteriology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, 852-8523, Nagasaki, Japan aut Kohno Shigeru Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki University School of Medicine, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut Digestive Diseases and Sciences Springer US; http://www.springer-ny.com 56/4(2011-04-01), 999-1006 0163-2116 56:4<999 2011 56 10620 https://doi.org/10.1007/s10620-010-1420-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10620-010-1420-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Matsumoto Ayako Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki University School of Medicine, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut NATIONALLICENCE 700 1- Isomoto Hajime Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut NATIONALLICENCE 700 1- Nakayama Masaaki Department of Bacteriology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, 852-8523, Nagasaki, Japan aut NATIONALLICENCE 700 1- Hisatsune Junzo Department of Bacteriology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, 852-8523, Nagasaki, Japan aut NATIONALLICENCE 700 1- Nishi Yoshito Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki University School of Medicine, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut NATIONALLICENCE 700 1- Nakashima Yujiro Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut NATIONALLICENCE 700 1- Matsushima Kayoko Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut NATIONALLICENCE 700 1- Kurazono Hisao Department of Applied Veterinary Medicine and Public Health, Obihiro University of Agriculture and Veterinary Medicine, 080-8555, Obihiro, Japan aut NATIONALLICENCE 700 1- Nakao Kazuhiko Department of Gastroenterology and Hepatology, Nagasaki University Hospital, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut NATIONALLICENCE 700 1- Hirayama Toshiya Department of Bacteriology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, 852-8523, Nagasaki, Japan aut NATIONALLICENCE 700 1- Kohno Shigeru Department of Molecular Microbiology and Immunology, Nagasaki University Graduate School of Biomedical Sciences, Nagasaki University School of Medicine, 1-7-1 Sakamoto, 852-8501, Nagasaki, Japan aut NATIONALLICENCE 773 0- Digestive Diseases and Sciences Springer US; http://www.springer-ny.com 56/4(2011-04-01), 999-1006 0163-2116 56:4<999 2011 56 10620 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer