The antisense block of CROC-1 gene expression makes cells grow slower

AIM:To establish FL-CROC- 1 ^- cell line in which CROC- 1 gene expression was blocked and study the role of CROC- 1 gene in cell growth. METHODS:The appropriate length cDNA fragment of the recently identified human gene CROC- 1 which encodes ubiquitin-conjugating enzyme like protein(Ubc-like protein) was cloned into the reconstructed eukaryotic expression vector pMAMneo-amp^- by antisense strategy. The recombinant plasmid which can express CROC- 1 antisense RNA was selected by restriction enzyme map analysis. The antisense expression recombinant plasmid pMAM-antiCROC- 1 was then transfected into human amnion FL cells by a modified calcium phosphate-mediated transfection procedure and selected with MEM medium containing 400 μg/mL geneticin. Finally ,the growth rate of the G418 resistant FL-CROC- 1 ^- cell line was determined.RESULTS:When the antisense inhibition of CROC-1 gene expression was induced by dexamethasone,the growth rate of the FL-CROC-1^- cel line was obviously slower as compared with that of the control FL cell and FL-MAMneo cell which was established by transfection of plasmid pMAMneo-amp^-(P<0.05).CONCLUSIONS:FL-CROC- 1 ^- cell line was successfully established in this study and the result of FL-CROC- 1 ^- cell growth suppression suggests that CROC- 1 gene encoded Ubc-like protein plays a positive regulation role in cell growth.