溶瘤腺病毒ATV联合顺铂协同抑制A 549细胞的研究

Study on synergistic suppression of A549 cells by a combination of ATV and cisplatin

目的探讨双特异性溶瘤腺病毒ATV与化疗药物顺铂联用对人肺癌细胞A 549治疗的增效减毒作用。方法将构建的溶瘤腺病毒ATV感染A 549细胞后进行Annexin V检测,分析ATV对肿瘤细胞的抑制方式;单用ATV、顺铂以及两者联合作用于人肺癌A 549细胞,采用MTS法检测细胞体外增殖情况,采用划痕试验、Transwell迁移试验和Transwell侵袭试验检测细胞的迁移侵袭情况;构建A 549皮下移植瘤裸鼠模型,单独注射重组腺病毒、顺铂以及重组腺病毒和顺铂联合使用,观察监测肿瘤生长情况,评价溶瘤腺病毒联合顺铂对肿瘤的体内抑制作用。结果双特异性重组腺病毒ATV主要通过诱导A 549细胞凋亡达到对细胞的杀伤;ATV联合顺铂体外杀伤A 549细胞具有一定的剂效和时效关系,且其抑制作用显著强于ATV及顺铂单独使用;ATV联合顺铂作用下,可使A 549细胞迁移侵袭能力降低,且能显著抑制小鼠皮下移植瘤生长,延长模型小鼠生存期。结论 ATV可诱导肿瘤细胞凋亡,ATV与化疗药物顺铂联用可增强对A549细胞生长及迁移侵袭能力的抑制作用,即具有抑制肺癌细胞A 549的增效减毒作用。

Objective To examine the synergistic platinum on the A549 human lung cancer cell line. effect of a dual cancer-specific oncolytic adenovirus （ATV） and cis Methods A549 cells were infected with ATV, and annexin V was detected. Inhibition of tumor cells by ATV was measured. Cell proliferation in vitro was assessed by infecting A549 cells with the oncolytic adenovirus ATV and treating those cells with cisplatinum, by infecting those cells with the oncolytic adenovirus ATV alone, or by treating those cells with cisplatinum alone. An MTS assay was performed. A wound scratch assay, Transwell migration assay, and Transwell invasion assay were used to detect the migration and invasion of A549 cells that were transfected with constructed adenoviral vectors. Nude mice bearing subcutaneously implanted tumors formed by A549 cells were treated with ATV alone or in combination with cisplatin. Tumor growth was monitored, and the inhibition of tumors by ATV combined with cisplatin was determined in vitro and in vivo. Results The dual cancerspecific oncolytic adenovirus ATV killed A549 cells by inducing apoptosis, and this action was dose and time-dependent. When A549 cells were treated with ATV combined with cisplatin, they had markedly diminished capacity for migration and invasion compared to cells that were treated with ATV or eisplatin alone. The use of ATV combined with cisplatin inhibited the growth of subcutaneously implanted tumors. Conclusion ATV can induce apoptosis of A549 cells. The combination of ATV and cisplatinum has a synergistic attenuating effect and can enhance inhibition of A549 cell growth and it can inhibit migration and invasion by those cells.