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目的:构建受辐射诱导的EGR-1启动子调控的携带人TRAIL基因的新型溶瘤腺病毒Ad-EGR-TRAIL,研究其联合放疗对宫颈癌细胞株HeLa S3的杀伤效果。方法:构建重组腺病毒Ad-EGR-TRAIL,用腺病毒Ad-GFP检测对HeLa S3细胞的感染效率。CCK-8法检测Ad-EGR-TRAIL组、单纯放疗组以及Ad-EGR-TRAIL联合放疗组对HeLa S3细胞的杀伤效应,同时观察它们对人正常宫颈细胞的作用。结果:成功构建腺病毒Ad-EGR-TRAIL,当MOI为100时,HeLa S3细胞的腺病毒感染效率最高。单纯Ad-EGR-TRAIL或放疗对HeLa S3细胞增殖的抑制率分别为(8.07±3.02)%和(23.02±4.03)%,Ad-EGR-TRAIL联合放疗对HeLa S3细胞增殖的抑制率达(79.77±9.15)%;同样的处理对正常宫颈细胞无明显抑制作用。结论:Ad-EGR-TRAIL联合放疗对宫颈癌细胞HeLa S3有显著的杀伤作用。
OBJECTIVE: To construct a novel oncolytic adenovirus Ad-EGR-TRAIL carrying human TRAIL gene under the regulation of EGR-1 promoter induced by radiation and study its killing effect on cervical cancer cell line HeLa S3. Methods: The recombinant adenovirus Ad-EGR-TRAIL was constructed and the infection efficiency of HeLa S3 cells was detected by adenovirus Ad-GFP. The killing effects of Ad-EGR-TRAIL group, radiotherapy alone group and Ad-EGR-TRAIL combined with radiotherapy group on HeLa S3 cells were detected by CCK-8 assay, and their effects on human normal cervical cells were also observed. Results: The adenovirus Ad-EGR-TRAIL was constructed successfully. When the MOI was 100, HeLa S3 cells had the highest adenovirus infection efficiency. The inhibitory rates of Ad-EGR-TRAIL or radiotherapy on the proliferation of HeLa S3 cells were (8.07 ± 3.02)% and (23.02 ± 4.03)%, respectively. The inhibition rate of HeLa S3 cells proliferation was 79.77% ± 9.15)%. The same treatment had no obvious inhibitory effect on normal cervical cells. Conclusion: Ad-EGR-TRAIL combined with radiotherapy has a significant killing effect on HeLa S3 cervical cancer cells.