title: GNB2 is a mediator of lidocaine-induced apoptosis in rat pheochromocytoma PC12 cells.
abstract: Lidocaine has been recognized to induce neurotoxicity. However, the molecular mechanism underlying this effect, especially the critical molecules in cells that mediated the lidocaine-induced apoptosis were unclear. In the present study, PC12 cells were administrated with lidocaine for 48h. Using MTT assay and flow cytometry, we found lidocaine significantly decreased the cell proliferation and S phases in PC12 cells with treatment concentrations, and significantly enhanced cell apoptosis with treatment concentrations. Two-dimensional gel electrophoresis (2-DE) analysis and LC-MS/MS were used to identification of protein biomarkers. Six proteins were identified. Among them, three were up-expressed including ANXA6, GNB2 and STMN1, other three were down-expressed including ubiquitin-linke protein 7 (UBL7), DDAH2 and BLVRB. Using qRT-PCR, we confirmed that lidocaine up-regulated the mRNA expression of STMN1, GNB2, ANXA6 and DDAH2, and found that the GNB2 had the largest change (about increased by 6.4 folds). The up-regulation of GNB2 by lidocaine was also validated by western blot. After transfected with 100μM GNB2-Rat-453 siRNA, the expression of GNB2 in PC12 cells was almost completely inhibited; and the cell proliferation and cells in S phases were significantly enhanced, cell apoptosis including both early apoptosis and later apoptosis were significantly reduced in the presence of 0.5mM lidocaine for 48h. Therefore, neuronal apoptosis was induced by lidocaine and this effect was mediated by GNB2. Further research is needed to assess the clinical relevance and exact mechanism of neuronal apoptosis caused by lidocaine.
authors.affiliation: Department of Anesthesiology, Guangzhou Women and Children's Medical Center, Guangzhou 510623, Guangdong, China. Electronic address: yonghtan@hotmail.com.
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Tan
authors.affiliation: Department of Anesthesiology, Guangzhou Women and Children's Medical Center, Guangzhou 510623, Guangdong, China. Electronic address: wangtans@hotmail.com.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Anesthesiology, Guangzhou Women and Children's Medical Center, Guangzhou 510623, Guangdong, China. Electronic address: zhaobaisong819@163.com.
authors.forename: Baisong
authors.initials: B
authors.lastname: Zhao
authors.affiliation: Department of Anesthesiology, Guangzhou Women and Children's Medical Center, Guangzhou 510623, Guangdong, China. Electronic address: yjsheh@yeah.net.
authors.forename: Yingjun
authors.initials: Y
authors.lastname: She
authors.affiliation: Department of Anesthesiology, Guangzhou Women and Children's Medical Center, Guangzhou 510623, Guangdong, China. Electronic address: bobjuan@sina.cn.
authors.forename: Xiaobao
authors.initials: X
authors.lastname: Bi
mesh_terms: D000581:Amidohydrolases / Q000235:genetics / Q000378:metabolism; D000704:Analysis of Variance; D000779:Anesthetics, Local / Q000633:toxicity*; D000818:Animals; D017304:Annexin A5 / Q000378:metabolism; D017317:Annexin A6 / Q000235:genetics / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects*; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D015180:Electrophoresis, Gel, Two-Dimensional; D044387:GTP-Binding Protein beta Subunits / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects*; D008012:Lidocaine / Q000633:toxicity*; D016716:PC12 Cells / Q000187:drug effects; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D050777:Stathmin / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D000779:Anesthetics, Local; D017304:Annexin A5; D017317:Annexin A6; D044387:GTP-Binding Protein beta Subunits; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D050777:Stathmin; C061082:Stmn1 protein, rat; D008012:Lidocaine; D000581:Amidohydrolases; C059999:dimethylargininase
keywords: 2-DE; Apoptosis; Cell cycle; GNB; Lidocaine