镉相关新基因TIF3的蛋白表达与细胞转化功能研究

雷毅雄; 陈家堃

镉相关新基因TIF3的蛋白表达与细胞转化功能研究

Study on protein expression and cell transformation of novel cadmium related gene TIF3

摘要

摘要:
目的探讨TIF3基因在镉细胞转化和致癌过程中的重要作用。
方法用细胞转染, Westernblot, 以及细胞转化等技术与方法, 对克隆化基因TIF3的生物学功能进行研究。
结果 TIF3cDNA以pcDNA3.1/V5-His-TOPO为表达载体所转染的CHO细胞和猴肾COS1细胞均可表达分子量为36000的TIF3编码蛋白质, 而非转染和单纯载体转染的对照细胞则无此蛋白质表达; 此外, TIF3cDNA转染NIH3T3细胞可导致TIF3蛋白质超额表达, 并与细胞转化密切相关。
结论镉的细胞转化和致癌作用至少部分是因TIF3基因的高表达所致。

Abstract:
Objective TEF-1δ, mouse translation elongation factor-1D, is a novel gene cloned from CdCl ₂-transformed BALB/c-3T3 cells.Its genbank accession number is AF304351.The objective is to test the biological functions of the gene in the cell transformation and car cinogenesis of cadmium.
Methods The transfection technique, western analysis and cell transformation assay were used.
Results Compared with untransfected and vector control, a 31 kDa protein was detected in Chinese hamster ovary(CHO)cells and in monkey kidney COS1 cells transfected with an expression vector(pcDNA 3.1/V5-His-TOPOV ector)containing TEF-1δcDNA.Furtheromore, transfection of NIH3T3 cells with an expression vector containing the TEF-1δcDNA resulted in overex pression of the encoded protein and this was associated with cell transformation.
Conclusion These findings demonstrated for the first time that the cell transformation and carcinogenesis induced by cadmium were due, at least in part, to the overexpression of TEF-1δ.It was probably a novel cadmium-responsive protooncogene.

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