shRNA干扰Rce1基因表达对口腔鳞癌细胞放射敏感性的影响  

Effect of shRNA interference Rce1 gene expression on the radiosensitivity of oral squamous cell carcinoma cells

作　　者：孙大为[1] 张云[2] 朱勇[2] SUN Da-wei;ZHANG Yun;ZHU Yong(Department of Stomatology,Baoji Central Hospital,Baoji 721008,Shaanxi Province,China;Department of Radiotherapy,Baoji Central Hospital,Baoji 721008,Shaanxi Province,China)

机构地区：[1]宝鸡市中心医院口腔科,陕西宝鸡721008 [2]宝鸡市中心医院放射治疗科,陕西宝鸡721008

出　　处：《中国口腔颌面外科杂志》2022年第2期123-128,共6页China Journal of Oral and Maxillofacial Surgery

摘　　要：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。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 lipo-some 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 soft-ware package was used for data analysis.RESULTS:Compared with the control group and shRNA-NC group,the inhibi-tion 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 de-creased,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.

关 键 词：口腔鳞癌 Ras转化酶1 放射敏感性 

分 类 号：R739.8[医药卫生—肿瘤]