干扰BAG-1表达对黑素瘤B16F10细胞放射敏感性的影响

目的: 明确BAG-1基因表达对黑素瘤B16F10细胞放射敏感性的影响及机制。方法: B16-F10细胞株分为未转染组(Control组)、阴性对照质粒转染组(NC-shRNA组) 和转染BAG-1-shRNA 组(BAG-1-shRNA组)。 克隆计数法计算克隆形成率和存活分数，描绘细胞存活曲线。流式细胞仪检测细胞凋亡率，Western blot检测BAG-1沉默对黑色素瘤细胞凋亡相关蛋白Caspase3、8、9、PARP表达。结果: X线8Gy 剂量照射后，Control组、NC-shRNA组和BAG-1-shRNA组细胞的存活率分别为1.9%，1.75%和0.32%，细胞凋亡率分别为7.1%，9.2% 和25.3%，差异具有统计学意义（均P＜0.05）。BAG-1-shRNA组调亡蛋白c-PARP及c-caspase 3、caspase 8、caspase 9水平分别为0.36, 0.37，0.35和0.30高于control组（0.10,0.04，0.11, 0.2）和NC-shRNA组（0.13, 0.12, 0.20, 0.26）。讨论: 干扰BAG-1基因表达可显著增高黑素瘤B16F10细胞对X放射线的敏感性，促进细胞凋亡可能是放射增敏的机制之一。

Effect of inhibiting of BAG-1 gene expression on radiosensitivity of melanoma B16F10 cells

Objective: To determine effect of inhibiting of BAG-1 gene expression on radiosensitivity of melanoma B16F10 cells and explore the mechanism. Methods: B16F10 cells were divided into three groups: control group, NC-shRNA group and BAG-1-shRNA group. Medical linear accelerator irradiation and clone counting method were used to calculate the clone formation rate and survival fraction, and the cell survival curve was delineated. The apoptosis rates were detected by flow cytometry. The expression level of apoptosis-related proteins Caspase 3, 8, 9, and c-PARP in melanoma cells were detected by Western blot. Results: After X-ray 8Gy dose irradiation, the survival rates of cells in the control group, NC-shRNA group and BAG-1-shRNA group linear were 1.9%, 1.75%, 0.32% and the apoptosis rate were 7.1%, 9.2% and 25.3% respectively, with significant differences (P<0.05). The levels of c-PARP, Caspase 3, 8, 9 were 0.36, 0.37，0.35 and 0.30 respectively, which were higher than those in the control group and NC-shRNA group, with significant differences (P<0.05). Conclusion: RNA interference of BAG-1 gene expression can significantly increase the sensitivity of melanoma cells to X-ray radiation, which may be associated with accelerating cells apoptosis.