title: Role of FAK in TNF-alpha/Cycloheximide-induced Apoptosis of SMMC-7721 Cells.
abstract: To explore the role of FAK in TNF-alpha/cycloheximide-induced apoptos is of human hepatocellular carcinoma cell line SMMC-7721, the FAK antisense plasmid was constructed and transfected into SMMC-7721 cells. Western blot assay was adopted to examine PKB level. Flow cytometry assay was used to detect apoptosis. It was shown that the SMMC-7721 cells were insensitive to TNF-alpha cytotoxicity, but they entered apoptosis quickly in the presence of cycloheximide and TNF-alpha. PKB was decreased during TNF-alpha/cycloheximide-induced apoptosis. No significant change of PKB level was found in the presence of TNF-alpha or cycloheximide, respectively, seeming that PKB level was closely correlated with apoptosis. When FAK was 60% reduced as a result of the transfection of SMMC-7721 cells with FAK antisense construct, the percentage of TNF-alpha/cycloheximide-induced apoptosis was enhanced at lower dose of TNF-alpha but decreased at higher dose of TNF-alpha, compared with the control. Correspondingly, the PKB level in FAK-down-regulated transfectants was lower at lower dose of TNF-alpha, but higher at higher dose of it. Therefore, FAK regulated TNF-alpha/cycloheximide-induced apoptosis in a biphase manner. This function might be related with PKB level.
authors.affiliation: Key Laboratory of Glycoconjugate Research, Ministry of Health, Department of Biochemistry, School of Medicine, Fudan University, Shanghai 200032, China. xlzha@shmu.edu.cn
authors.forename: Yi
authors.initials: Y
authors.lastname: Fang
authors.forename: Jia-Wei
authors.initials: JW
authors.lastname: Jin
authors.forename: Xi-Liang
authors.initials: XL
authors.lastname: Zha