反义阻断hREV3基因表达对细胞生长及MNNG敏感性的影响

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

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 (2 93细胞 )细胞系 (2 93 -hREV3-)。以细胞记数方法检测该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 (2 93-hREV3-)与对照组母本 2 93细胞和转染载体DNA的 2 93细胞相比在生长速率上无明显差异 ,但对烷化剂MNNG的细胞毒作用较为敏感。结论 :hREV3基因编码蛋白并非细胞增殖所必需 。

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.