shRNA干扰Rce1基因表达对口腔鳞癌细胞放射敏感性的影响

目的:观察shRNA干扰Ras转化酶1（Rce1）基因表达对口腔鳞癌（OSCC）细胞放射敏感性的影响。方法:取对数期CAL27细胞,随机分为Control组、shRNA-NC组、shRNA-Rce1组。Control组不处理,shRNA-NC组、shRNA-Rce1组分别采用脂质体转染法转染shRNA-NC、shRNA-Rce1表达载体。采用二甲基噻唑（MTT）法检测不同放射剂量（2、4、6、8、10 Gy）射线照射对CAL27细胞增殖抑制率的影响,计算放射对细胞的半数抑制剂量。利用AnnexinV-FITC/PI双染法检测细胞凋亡率,Western免疫印迹法检测细胞B细胞淋巴瘤2（Bcl-2）、存活蛋白（Survivin）、半胱天冬氨酸蛋白酶3（Caspase-3）蛋白表达量。采用SPSS 19.0软件包对数据进行统计学分析。结果:与Control组、shRNA-NC组相比,不同放射剂量对shRNA-Rce1组CAL27细胞的增殖抑制率显著升高,半数抑制剂量显著降低（P<0.05）;与Control组、shRNA-NC组相比,shRNA-Rce1组凋亡率显著升高（P<0.05）;与Control组、shRNA-NC组相比,shRNA-Rce1组Bcl-2、Survivin蛋白表达量显著降低,Caspase-3蛋白表达量显著升高（P<0.05）。结论:shRNA干扰Rce1基因表达可增强OSCC细胞放射敏感性,降低半数抑制剂量,促进细胞凋亡,并减少凋亡抑制蛋白Bcl-2、Survivin表达,增加凋亡蛋白Caspase-3表达。

Effect of shRNA interference Rce1gene expression on the radiosensitivity of oral squamous cell carcinoma cells

PURPOSE: To observe the effect of shRNA interference with Ras convertase 1 (Rce1) gene expression on the radiosensitivity of oral squamous cell carcinoma (OSCC) cells. METHODS: CAL27 cells in log phase were taken and randomly divided into control group, shRNA-NC group and shRNA-Rce1 group. The control group was not treated, while shRNA-NC group and shRNA-Rce1 group were transfected with shRNA-NC and shRNA-Rce1 expression vector by liposome transfection method, respectively. Dimethylthiazole (MTT) method was used to detect the proliferation inhibition rate of different radiation doses (2, 4, 6, 8, 10 Gy) on CAL27 cells, and the half-dose radiation inhibitory amount on the cells was calculated. AnnexinV-FITC/PI double staining was used to detect the apoptosis rate. Western blot was used to detect the expression levels of cellular B-cell lymphoma-2(Bcl-2), Survivin and Caspase-3 (Caspase-3) protein. SPSS 19.0 software package was used for data analysis. RESULTS: Compared with the control group and shRNA-NC group, the inhibition rate of different radiation doses on CAL27 cell proliferation in the shRNA-Rce1 group were significantly increased, while the half inhibition dose was significantly decreased(P<0.05). Compared with the control group and shRNA-NC group, the apoptotic rate in the shRNA-Rce1 group was significantly increased (P<0.05). Compared with the control group and shRNA-NC group, the expression of Bcl-2 and Survivin protein in the shRNA-Rce1 group was significantly decreased, while the expression of Caspase-3 protein was significantly increased (P<0.05). CONCLUSIONS: shRNA interference with Rce1 gene expression can enhance the radiosensitivity of OSCC cells, reduce half inhibition dose, promote cell apoptosis, reduce the expression of apoptosis-inhibiting proteins Bcl-2 and Survivin, and increase the expression of apoptotic protein Caspase-3.