Effect on Gap Junctional Intercellular Communication by TPT in WB-F344 Rat Liver Epithelial Cells

• Main Authors: I-Hui Chen, 陳怡橞
• Other Authors: Jin-Lian Tsai
• Format: Others
• Language: zh-TW
• Published: 2007
• Online Access: http://ndltd.ncl.edu.tw/handle/20276410418208959975

碩士 === 高雄醫學大學 === 職業安全衛生研究所碩士班 === 95 === Organotin compounds (OTCs) are used extensively as stabilizers in the production of plastic,agricultural pesticides, antifoulant paints and wood preservation. Owing to their structure are similar to hormone. OTCs are category Environmental Hormone induce toxicity in animals and human such as interrupt normal metabolism and neurotoxicity. There are a few studies about Gap Junctional Intercellular Communication (GJIC) of cells by OTCs. Therefore, this study discussed the effect on GJIC in WB-F344 rat liver epithelial cells by TPT. According to the MTT assay, cell availabilities of WB-F344 cell were degression when treaded with high concentration TPTC (IC50=59 ppb) and had Dose & Time dependent. And WB-F344 cells Colony Forming Efficiency were reversed of TPTC dose when exposured 9 days. During the GJIC experimentation,WB-F344 cells GJIC were block when treated 1.5 ppm TPTC in 30 minutes. The diffusion length were decreased gradually when exposured 1.5 ppm TPTC 15、30、45、60 minutes. The GJIC were recover when WB-F344 cells treated with PD98059(ERK inhibitor ) and LY294002（PI 3-kinase inhibitor） previously, but treated with GF109203X hydrochloride (Protein Kinase C inhibitor)were not. TPTC increased extra- and intra-cellular calcium in WB-F344 cells In the Immunostain analysis & Western Blot experimentation, the expression of GAPDH、β-caternin were the same, but E-cadherin in WB-F344 cells with TPTC were strong than control. The fluorescence of Cx43 in WB-F344 cells with TPTC was decrease and Cx43 phosphoration be found in Western Blot experimentation . According to those experimentations, the mechanism of WB-F344 cells GJIC block was TPTC destroyed the function of protein on the cell. Because of membranes that intracellular calcium concentration change, PI3-kinase and MEK pathway were be activate and caused Cx43 phosphoration.