目的:探讨重组溶瘤腺病毒Ad-Apoptin-hTERTp-E1A(ATV)对荧光素酶标记人肺癌细胞（A549-luc）和人肺癌细胞（A549）的体外抑制作用差异。方法：利用ATV分别感染A549 细胞和A549-luc 细胞，通过WST-1 和结晶紫染色法确定ATV的抑制作用的差异性，通过Hoechst 和Annexin V-FITC/PI 染色验证ATV的抑制方式。结果：ATV对A549 和A549-luc 细胞均具有明显抑制作用（P<0.05）。ATV在24、48、72 h 对两种细胞均有明显抑制作用（P<0.05 或P<0.01），且72 h 抑制率最强；剂量分别为1、10 和100 MOI的ATV对两种细胞均有抑制作用，且100 MOI抑制率最高。A549 细胞和A549-luc 细胞感染ATV后，均呈现核碎裂典型浓染和边集的凋亡现象，且凋亡率随时间的增加而增大，具有显著的时间效应（P<0.05 或P<0.01），且凋亡率均在72 h 达到峰值。结论：荧光素酶的插入没有明显改变ATV对A549-luc 细胞的抑制作用和抑制方式，ATV是通过诱导A549-luc 细胞和A549细胞凋亡从而达到对其显著性体外抑制作用.

Objective: To investigate the inhibitory effect of recombinant oncolytic adenovirus Ad-Apoptin-hTERTp-E1A (ATV) on luciferase-labeled human lung cancer cells (A549-luc) and human lung cancer A549 cells, and to compare the differences in the inhibitory effect on two cell lines. Methods: ATV was used to infect A549-luc cells and A549 cells respectively. WST-1 and crystal violet staining were used to determine the difference in the inhibitory effect of ATV. Hoechst and Annexin V-FITC/PI staining were used to verify the inhibition mode of ATV. Results: WST-1 and crystal violet staining showed that ATV had significant inhibitory effect on both A549-luc and A549 cells (P<0.05). ATV showed significant inhibitory effect on both cells at 24, 48 and 72 h (P<0.05 or P<0.01), and reached the peak at 72 h; ATV at concentrations of 1, 10 and 100 MOI all showed inhibitory effect on both cells, and reached the peak at 100 MOI.Hoechst staining showed that A549-luc cells and A549 cells infected with ATV showed typical nuclear fragmentation and marginal set.The results of Annexin V-FITC/PI Flow cytometry showed that ATV infection resulted in apoptosis of A549-luc and A549 cells, which was in a time-dependent manner and reached the peak at 72 h(P<0.05 or P<0.01). Conclusion: Insertion of luciferase didn’t significant-ly change the inhibitory effect and inhibitory mode of ATV on A549-luc cells. ATV exerted its in vitro inhibitory effect on A549-luc and A549 cells by inducing cell apoptosis.

国家重点研发计划重要新发突发病体防治处治技术与研究资助项目（No.2016YFC1200900）；长春市重大科技攻关资助项目（No.16ss11）；国家科学技术重大新药创制专项（No.2018ZX09301053-004-001）