Establishment of cell lines whose hREV3 gene expression was inhibited by transfection of antisense RNA expression plasmids and their biological characteristics / 中国病理生理杂志

ABSTRACT

AIM: to establish the cell lines whose hREV3 gene expression was blocked by antisense RNA and observe their characteristis of cell growth rate and N - methyl - N' - nitrosoguanidine (MNNG) sensitivi- ty . METHODS: With modified calcium phosphate - DNA coprecipitation method the eukaryocytic expression plasmid expressing antisense fragment of hREV3,pBK - RSV - hREV3- and pMAMneo - amp hREV3 were transfected into human embryo kidney cell line of HEK - 293. After G418 selection, cell lines of 293 - B - hREV3- and 293 - M hREV3- were established. By cell counting method, the cell growth rate and MNNG sensitivity of these cell lines were characterized. RESULTS: No change of cell growth rates of these cell lines was observed whether hREV3 gene expres- sion was blocked by either the persistent or induced expression of the antisense hREV3 RNA. While the sensitivity of these cell lines to MNNG was somewhat elevated, as compared with their parent cell line 293 and the cell lines trans- fected with vector DNAs. CONCLUSION: The gene product of hREV3 was not essential for the cell growth, but it may play a role in the DNA repair functions of the cells after exposure to DNA damaging agents.