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李丽, 冯俊, 王朝莉, 等. 反义ATM核苷酸转染对放射线照射后喉鳞癌细胞凋亡影响的体外研究[J]. koko体育app 学报（医学版）, 2013, 44(3): 379-382.
引用本文: 李丽, 冯俊, 王朝莉, 等. 反义ATM核苷酸转染对放射线照射后喉鳞癌细胞凋亡影响的体外研究[J]. koko体育app 学报（医学版）, 2013, 44(3): 379-382.
LI Li, FENG Jun, WANG Chao-li, et al. The Apoptosis Effect of Antisense Oligodeoxynucleotides Targeting ATM on the Laryngeal Squamous Cell Carcinoma Treated with Radiation[J]. Journal of Sichuan University (Medical Sciences), 2013, 44(3): 379-382.
Citation: LI Li, FENG Jun, WANG Chao-li, et al. The Apoptosis Effect of Antisense Oligodeoxynucleotides Targeting ATM✃ on the Laryngeal Squamous Cell Carcinoma Treated with Radiation[J]. Journal of Sichuan University (Medical Sciences), 2013, 44(3): 379-382.

反义ATM核苷酸转染对放射线照射后喉鳞癌细胞凋亡影响的体外研究

The Apoptosis Effect of Antisense Oligodeoxynucleotides Targeting ATM on the Laryngeal Squamous Cell Carcinoma Treated with Radiation

  • 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18 h,各组细胞分别接受0、2、4、6、8 Gy照射,24 h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4 Gy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。  
    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.  
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