title: Helicobacter pylori infection combined with DENA revealed altered expression of p53 and 14-3-3 isoforms in Gulo-/- mice.
abstract: Unlike most other mammals, human bodies do not have the ability to synthesize vitamin C inside of their own bodies. Therefore, humans must obtain vitamin C through daily diet. Gulo(-/-) mice strain is known with deficiency, in which vitamin C intake can be controlled by diet like human, and would be valuable for investigating the molecular mechanism of various diseases. In the present study, we established Gulo(-/-) mice model and investigated the differentially expressed proteins in stomach tissue of Gulo(-)(/-) mice after Helicobacter pylori-infected, and followed by DENA, using immunohistochemistry and proteomic approach. The results of immunohistochemistry analysis of stomach tissue showed that the tumor suppressor, p53 protein, expression was significantly decreased (p<0.05) but not messenger RNA (mRNA) transcriptional level, and 14-3-3 ε, 14-3-3 δ, Ki-67 and cleaved caspase 3 expressions were significantly increased (p<0.05) by H. Pylori infection, and followed by DENA treatment in Gulo(-/-) mice. Moreover, knockdown of 14-3-3 isoforms (14-3-3 ε, 14-3-3 σ, 14-3-3 ζ and 14-3-3 η) were significantly increased sub-G1 phase (characteristics of apoptosis) in AGS cells and, phenotypic changes like cell shrinkage, density and cleaved nuclei were also observed. Proteome analyses showed that 14-3-3 σ, 14-3-3 η, and tropomyosin alpha-1 chain were down-regulated, and Hspd1 protein and HSC70 were up-regulated after H. Pylori-infection, and followed by DENA. The combined results of immunohistochemistry and proteomic analysis suggest that H. pylori altered the p53 and 14-3-3 isoforms expression and DENA further enhanced the H. pylori effect, which might be involved in carcinogenesis and metastasis of gastric cancer on Gulo(-/-) mice.
authors.affiliation: Research Institute of Life Science and College of Veterinary Medicine, Gyeongsang National University, 900 Gajwadong, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Arulkumar
authors.initials: A
authors.lastname: Nagappan
authors.forename: Hyeon Soo
authors.initials: HS
authors.lastname: Park
authors.forename: Kwang Il
authors.initials: KI
authors.lastname: Park
authors.forename: Gyeong Eun
authors.initials: GE
authors.lastname: Hong
authors.forename: Silvia
authors.initials: S
authors.lastname: Yumnam
authors.forename: Ho Jeong
authors.initials: HJ
authors.lastname: Lee
authors.forename: Mun Ki
authors.initials: MK
authors.lastname: Kim
authors.forename: Eun Hee
authors.initials: EH
authors.lastname: Kim
authors.forename: Won Sup
authors.initials: WS
authors.lastname: Lee
authors.forename: Wang Jae
authors.initials: WJ
authors.lastname: Lee
authors.forename: Myung Je
authors.initials: MJ
authors.lastname: Cho
authors.forename: Woo Kon
authors.initials: WK
authors.lastname: Lee
authors.forename: Chung Kil
authors.initials: CK
authors.lastname: Won
authors.forename: Jae Hyeon
authors.initials: JH
authors.lastname: Cho
authors.forename: Gon Sup
authors.initials: GS
authors.lastname: Kim
mesh_terms: D048948:14-3-3 Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000477:Alkylating Agents / Q000633:toxicity*; D000818:Animals; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D018834:Chaperonin 60 / Q000378:metabolism; D004052:Diethylnitrosamine / Q000633:toxicity*; D005753:Gastric Mucosa / Q000378:metabolism; D005786:Gene Expression Regulation; D050883:HSC70 Heat-Shock Proteins / Q000378:metabolism; D016481:Helicobacter Infections / Q000378:metabolism* / Q000473:pathology; D016480:Helicobacter pylori; D006801:Humans; D019394:Ki-67 Antigen / Q000378:metabolism; D050544:L-Gulonolactone Oxidase / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D024101:Mitochondrial Proteins / Q000378:metabolism; D020033:Protein Isoforms / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D014335:Tropomyosin / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
chemical_list: D048948:14-3-3 Proteins; D000477:Alkylating Agents; D018834:Chaperonin 60; D050883:HSC70 Heat-Shock Proteins; C557066:Hspd1 protein, mouse; D019394:Ki-67 Antigen; D024101:Mitochondrial Proteins; D020033:Protein Isoforms; D034741:RNA, Small Interfering; C490774:Tpm1 protein, mouse; D014335:Tropomyosin; D016159:Tumor Suppressor Protein p53; D004052:Diethylnitrosamine; D050544:L-Gulonolactone Oxidase; D053148:Caspase 3
keywords: 14-3-3 Isoforms; Diethylnitrosamine; Gastric cancer; Gulo(−/−) mice; Helicobacter pylori; Proteome analysis