反义阻断hREV3基因表达对细胞生长及MNNG敏感性的影响

目的：应用反义核酸阻断技术研究ｈＲＥＶ３基因对细胞生长和ＭＮＮＧ敏感性的影响。方法：酶切法构建表达反义ｈＲＥＶ３基因片段的真核细胞表达质粒；以改良的磷酸钙－ＤＮＡ共沉淀法建立ｈＲＥＶ３基因表达被阻断的人胚肾细胞（２９３细胞）细胞系（２９３－ｈＲＥＶ３＾－）。以细胞记数方法检测核细胞系的生长速率及不同浓度的烷化剂ＭＮＮＧ对细胞的毒作用。结果：ｈＲＥＶ３基因表达被阻断的人胚肾细胞（２９３－ｈＲＥＶ３＾

The Role of Antisense-blocked hREV3 Gene Expression in the Cell Growth and MNNG Sensitivity

Objective: To study the role of hREV3 gene in the cell growth and MNNG sensitivity of the cell lines with the technique of antisense nucleic acid. Methods: To construct the eukaryotic expression plasmid by digesting with restriction enzymes. And to establish the human embryo kidney cell lines whose hREV3 gene expression was blocked by the modified calcium phosphate-DNA coprecipitation method. By cell counting method, the cell growth rate and MNNG sensitivity of these cell lines were characterized. Results: Compared with their parent cell line 293 and the cell line transfected with vector DNAs, no change was found in the cell growth rate, but the sensitivity of MNNG was somewhat elevated. Conclusion: The gene product of hREV3 is 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.