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干扰素β(IFN-β)和肿瘤坏死因子相关凋亡诱导配体(TRAIL)是有效抗癌药物。腺相关病毒(AAV)为目前最有应用前景的基因转移载体之一。利用AAV携带IFN-β和TRAIL基因并置于hTERT启动子控制下分别构建成肿瘤靶向病毒AAV-hTERT-IFN-β和AAV-hTERT-TRAIL,且单个IFN-β或TRAIL基因治疗发挥了一定的抗癌效果。将AAV-hTERT-IFN-β和AAV-hTERT-TRAIL进行联合,旨在研究其对A549肺癌细胞体内外的生长抑制效应。ELISA法检测了AAV-hTERT-IFN-β感染A549细胞后分泌型IFN-β的表达;MTT法检测AAV-hTERT-IFN-β联合AAV-hTERT-TRAIL对肿瘤细胞的生长抑制作用;凋亡细胞染色和流式细胞仪分别检测了AAV-hTERT-IFN-β、AAV-hTERT-TRAIL及其联合对A549细胞的凋亡效应;进一步评价了联合AAV-hTERT-IFN-β和AAV-hTERT-TRAIL对A549裸鼠移植瘤的抑癌效果。结果显示,联合治疗优于任一单独治疗并且导致了增强的肿瘤细胞毒性和凋亡诱导效应。更进一步显示,联合AAV-hTERT-IFN-β和AAV-hTERT-TRAIL治疗发挥了重要的抑制裸鼠移植瘤效果甚至消除全部移植瘤,为探究IFN-β和TRAIL联合抗癌的分子机制奠定了基础。
Interferon beta (IFN-beta) and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) are potent anti-cancer drugs. Adeno-associated virus (AAV) is one of the most promising gene transfer vectors at present. The AAV-hTERT-IFN-β and AAV-hTERT-TRAIL were constructed by AAV carrying IFN-β and TRAIL gene respectively under the control of hTERT promoter, and the treatment of single IFN-β or TRAIL gene was certain Anti-cancer effect. AAV-hTERT-IFN-β and AAV-hTERT-TRAIL were combined to study their growth inhibitory effects on A549 lung cancer cells in vitro and in vivo. The expression of secreted IFN-βin A549 cells infected by AAV-hTERT-IFN-β was detected by ELISA. The growth inhibition of AAV-hTERT-IFN-β combined with AAV-hTERT-TRAIL was detected by MTT assay. The effects of AAV-hTERT-IFN-β, AAV-hTERT-TRAIL and their combination on the apoptosis of A549 cells were detected by flow cytometry, staining and flow cytometry. AAV-hTERT-IFN-β and AAV-hTERT- Tumor Suppressing Effect on A549 Nude Mouse Transplanted Tumor. The results show that combination therapy is superior to either single treatment and results in enhanced tumor cytotoxicity and apoptosis-inducing effects. Further, the combination of AAV-hTERT-IFN-β and AAV-hTERT-TRAIL therapy exerted an important inhibitory effect on nude mice xenografts and even eliminated all transplanted tumors, laying a foundation for exploring the molecular mechanism of IFN-β and TRAIL in combating cancer basis.