HR24L基因过表达抑制细胞凋亡

Overexpression of HR24L gene inhibits apoptosis

目的 :研究人类DNA损伤修复基因HR2 4L与细胞凋亡的关系。方法 :PCR方法扩增HR2 4L基因开放阅读框序列 ,克隆入逆转录病毒载体pDOR neo ,转染HeLa细胞 ,筛选阳性克隆 ;Southern印迹验证HR2 4L基因的整合状况。Northern印迹检查HR2 4LmRNA的细胞内表达水平。过氧化氢、丁酸钠及血清饥饿诱导凋亡 ,流式细胞计数仪检测凋亡比例 ,电泳观察DNALadder,Western印迹检测凋亡相关蛋白质的表达。结果 :获得转染有HR2 4L基因的HeLa细胞。与HeLa细胞相比 ,HeLa HR2 4L细胞HR2 4LmRNA表达明显上升。HR2 4L基因过表达导致Bcl 2的表达升高 ,抑制caspase 3和Bax的表达 ,而对p53却无明显的影响。HR2 4L基因过表达抑制过氧化氢和血清饥饿诱导的细胞凋亡 ,但对丁酸钠诱导的细胞凋亡却无抑制作用。结论 :HR2 4L基因过表达抑制细胞凋亡。

Objective: To study the effect of human DNA repair gene HR24L on apoptosis. Methods: The open reading frame of HR24L was amplified by PCR and was cloned into the retroviral vector pDOR neo. Transfection of HeLa cells was performed with the recombinant vector. Apoptosis was induced by H 2O 2, sodium butyrate and serum starvation. FACS and DNA agarose gel electrophoresis were used to observe apoptosis. Western blot analysis examined apoptosis related proteins. Results: HeLa cells were successfully transfected with HR24L gene, and overexpression of HR24L gene resulted in ascendance of Bcl 2, inhibited the expression of caspase 3 and Bax and suppressed the apoptosis caused by H 2O 2, and serum starvation, but had no effect on the expression of p53 and the apoptosis caused by sodium butyrate. Conclusion: Overexpression of the human repair gene HR24L can inhibit apoptosis.