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目的:探讨抑制Nbs1基因的表达对喉鳞癌细胞放射敏感性的影响。方法:用ADV1包装的干扰质粒(ADV1-Nbs1)和空载腺病毒(ADV1-NC)转染Hep-2细胞,将细胞分为正常对照组、ADV1-Nbs1转染组、ADV1-NC转染组,上述各组又分为X射线照射组和未照射组。RT-PCR和Western Blot检测Nbs1基因的表达水平,MTT法描绘出各组增殖曲线,流式细胞仪检测细胞周期,RT-PCR法测定各组相对端粒长度。结果:RT-PCR和Western Blot结果显示ADV1-Nbs1转染组的Nbs1基因的表达水平明显低于正常对照组和ADV1-NC转染组。ADV1-Nbs1转染联合X射线放射组抑制增殖最显著(P<0.05)。对比正常对照组(11.8%),ADV1-Nbs1转染组和ADV1-Nbs1转染联合放射组G_2/M期百分比均有增高,分别为34.2%(P<0.05)、25%(P<0.05)。ADV1-Nbs1转染组端粒长度较正常组缩短约57%(P<0.05),ADV1-Nbs1转染组同ADV1-Nbs1转染联合放射组端粒长度无差异(P>0.05)。结论:干扰MRN复合体功能可有效使Hep-2细胞的放射敏感性提高。
Objective: To investigate the effect of inhibiting the expression of Nbs1 gene on radiosensitivity of laryngeal squamous cell carcinoma. METHODS: Hep-2 cells were transfected with ADV1-packaged interfering plasmid (ADV1-Nbs1) and adenovirus vector (ADV1-NC). The cells were divided into normal control group, ADV1-Nbs1 transfection group and ADV1- Group, the above groups are divided into X-ray irradiation group and non-irradiation group. The expression of Nbs1 gene was detected by RT-PCR and Western Blot. The proliferation curve of each group was plotted by MTT method. The cell cycle was detected by flow cytometry. The relative telomere length was determined by RT-PCR. Results: The results of RT-PCR and Western Blot showed that the expression level of Nbs1 gene in ADV1-Nbs1 transfected group was significantly lower than that in normal control group and ADV1-NC transfected group. ADV1-Nbs1 transfection combined with X-ray radiation inhibited proliferation most significantly (P <0.05). Compared with the normal control group (11.8%), the percentage of G 2 / M phase were increased in ADV1-Nbs1 transfection group and ADV1-Nbs1 transfection combined with radiotherapy group (34.2%, P <0.05) . The telomere length in ADV1-Nbs1 transfected group was 57% shorter than that in normal group (P <0.05). There was no difference in telomere length between ADV1-Nbs1 transfection group and ADV1-Nbs1 transfection group (P> 0.05). Conclusion: The interference of MRN complex function can effectively improve the radiosensitivity of Hep-2 cells.