c-ski对大鼠皮肤成纤维细胞抗辐射作用的影响

Radioprotective effect of c-ski on rat skin fibroblast in vitro

目的研究原癌基因c ski对大鼠皮肤成纤维细胞抗辐射作用的影响,并探讨其可能的机理。方法采用Annexin V FITC PI标记的方法,通过流式细胞仪检测2～8Gy的软X线照射对培养的大鼠皮肤成纤维细胞凋亡的影响;观察转染c ski基因后细胞凋亡的变化;并采用Westernblot检测c ski对凋亡蛋白Bax、Bcl2表达的影响。结果软X线照射以剂量依赖的方式增加细胞凋亡,并且随着时间的延长进一步增加,大约在36h时达到峰值。转染c ski基因使4Gy照射24h后的成纤维细胞凋亡率显著下降。转染c ski基因48h后,Bax的表达和对照组差异无统计学意义,而Bcl2的表达显著增加。结论c ski可降低成纤维细胞的辐射敏感性,促进Bcl-2的表达可能是其机理之一。

Objective To examine radioprotective effect of c-ski on rat skin fibroblast in vitro and explore its possible mechanism. Methods The effect of soft X-ray irradiation at dose varied from 2 to 8 Gy on cell apoptosis in rat skin fibreblast were determined by flow cytometry with Annexin-V-FITC-Pl labelling. The effect of c-ski gene transfection on cell apoptosis was evaluated after soft X-ray irradiation of 4 Gy, The protein expressions of Bax and Bcl-2 after c-ski gene transfection were measured with the Western blot method. Results Soft X-ray irradiation increases cell apoptosis, and the increase is proportional to the irradiation （lose. Apoptosis ratio increases with time since the irradiation, and reaches its peak at 36h after the irradiation, c-ski gene was observed to markedly decrease apoptosis index at 24 h after soft X-ray irradiation of 4 Gy compared to the control group, significant increase of the protein expression of Bcl-2 was observed. C-ski gene was found no significant effect on the protein expression of Bax. Conclusion c-ski gent can decrease radiation sensitivity of skin fibroblast, promoting Bcl-2 protein expression is one of its possible mechanism for this radioprotective effects.