【摘要】 目的设计共济失调毛细血管扩张症突变基因(ATM)反义多核苷酸作用于喉鳞状细胞癌(简称喉鳞癌)细胞以减少ATM表达,进而分析在体外其对放射线照射后喉鳞癌细胞凋亡的影响。方法实验分为反义ATM核苷酸(AS-Lipo)、正义ATM核苷酸(Sen-Lipo)、错配ATM核苷酸(Mis-Lipo)、Lipo与Hep-2组。用实时荧光定量PCR对各组Hep-2细胞的ATM mRNA进行分析。转染后18h,各组细胞分别接受0、2、4、6、8Gy照射,24h后计算细胞存活分数(SF)检测Hep-2细胞放射线照射后的生存能力。选择4Gy的照射剂量,用流式细胞仪分析各组Hep-2细胞的凋亡情况。结果 AS-Lipo、Sen-Lipo、Mis-Lipo、Lipo与Hep-2组的ATM mRNA相对表达量以AS-Lipo组最低,为(11.03±2.51)%(P<0.05)。在同等剂量的放射线照射下,AS-Lipo组的SF最低,与其他组比较差异有统计学意义(P<0.05)。流式细胞仪检测发现,AS-Lipo组的凋亡率为(30.7±1.31)%,高于其他各组(P<0.05)。结论在体外,ATM反义多核苷酸降低了喉鳞癌细胞中ATM mRNA的表达,同时增加了放射线照射后喉鳞癌细胞的凋亡。更多还原

【Abstract】 Objecitve To designed antisense oligodeoxynucleotides(AS-ODNs) to reduce the expression of ATM and to study its effect on the apoptosis of Hep-2(human epidermoid laryngeal carcinoma) cells treated with radiation in vitro.Methods Experiment was divided into AS-Lipo,Sen-Lipo,Mis-Lipo,Lipo and Hep-2 group.The expression of ATM mRNA in Hep-2 cells was examined by real-time quantitative PCR.About 18 hours after transfection,they were irradiated simultaneously with different doses of X-ray radiation(0,2,4,6,and 8 Gy) respectively.Clonogenic survival assay was carried out to detect the survival ability of Hep-2 cells after irradiation.After exposed to 4 Gy radiation,flow cytometry was carried out to analyze the cell apoptosis.Results The relative ATM mRNA expression in Hep-2 cells treated with ATM AS-ODNs was decreased to(11.03±2.51)% which was much lower than that of untreated cells(P<0.05).After irradiation,the survival fraction(SF) of cells treated with ATM AS-ODNs was lower than that of other groups at the same dose of radiation.There was statistical significance between the group treated with ATM AS-ODNs and other groups(P<0.05).The apoptotic rate for the group irradiated with ATM AS-ODNs was(30.7±1.31)%,which was significantly higher than that of others(P<0.05).Conclusion AS-ODNs of ATM reduce ATM mRNA expression and enhance Hep-2 cells apoptosis to radiation in vitro.更多还原