Ad-ING4抑制人A549肺癌细胞及其裸鼠移植瘤的生长

Inhibitory effects of adenovirus-mediated ING4 on growth of human A549 lung cell and lung carcinoma xenografts in nude mice

目的研究腺病毒介导的ING4基因(Ad-ING4)对肺癌体内外的抑制作用及其潜在的分子机制。方法体外采用50 MOI的Ad-ING4感染A549肺癌细胞,并用MTT法检测Ad-ING4对A549细胞的抑制作用,流式细胞仪检测肿瘤细胞的凋亡率。采用A549细胞株建立人肺癌裸鼠模型,瘤体局部注射干预用药,每隔5天测量一次肿瘤体积,并计算瘤重抑瘤率;瘤块免疫组化检测ING4、生存素(survivin)、CD34、HIF-1等基因的表达。结果Ad-ING4感染A549细胞抑制率可达47.62%;72 h凋亡率为18.5%;荷瘤裸鼠经治疗后,Ad-ING4组和顺铂(DDP)组的肿瘤体积、瘤重均明显缩小,其抑瘤率分别达到42.43%和46.47%,DDP组出现不良反应;免疫组化检测Ad-ING4组和DDP组与对照组比较,生存素、CD34、HIF-1的表达下调(P<0.01)。结论Ad-ING4对人A549肺癌细胞及其裸鼠移植瘤的生长具有明显的抑制作用。其抑瘤机制可能与下调生存素、CD34、HIF-1的基因表达进而诱导肿瘤细胞凋亡和抑制肿瘤血管形成等有关。

Objective To study the inhibitory effects and the possible mechanisms of adenovirus-mediated ING4（Ad-ING4） on lung carcinoma in vitro and vivo.Methods Human A549 lung cells were infected by Ad-ING4 at 50 MOI.The growth inhibition of Ad-ING4 on A549 cells was detected by MTT.The apoptosis of tumor cells was detected by flow cytometry.The human lung carcinoma xenografts model was established with A549 cell in nude mice.Mice were progressed multi-points injection in tumor,the gross tumor volumes were measured every 5 days.The weights of tumor were measured and the ratios of tumor-suppression were calculated.The expression of ING4,survivin,CD34 and HIF-1 in tumor sample were detected by immunohistochemistry.ResultsThe inhibition ratio was 47.62%.The apoptotic rate was 18.5% at 72 hour.After treated,the tumor volume and weight in groups of Ad-ING4 and cisdiaminodichloplatin（DDP） were reduced,and the inhibitory rates were 42.43% and 46.47%,respectively.DDP-treated group showed adverse reaction.According to immunohistochemistry,survivin,CD34 and HIF-1 were lower or less（P0.01）.Conclusion Ad-ING4 inhibits the growth of human A549 lung cells and lung carcinoma in nude mice significantly,which may be related to multi-pathways such as down-regulation the expressions of survivin,CD34 and HIF-1 to induce apoptosis and inhibit angiogenesis.