靶向LASP1基因的干扰腺病毒对非小细胞肺癌的抗肿瘤作用

Antitumor effects of interfering adenovirus targeting LASP1 gene on non-small cell lung cancer

目的探讨靶向LIM和SH3蛋白1基因(LASP1)干扰腺病毒(Ad-LASP1-shRNA)对非小细胞肺癌(NSCLC)的抗肿瘤作用。方法采用实时荧光定量聚合酶链反应(qPCR)及Western blot检测NSCLC组织中LASP1表达水平;应用Ad-LASP1-shRNA感染A549细胞48 h;采用qPCR、Western blot分析转染后A549细胞中mRNA及蛋白的表达水平;划痕实验和Transwell法检测转染后A549的迁移与侵袭能力;建立A549移植瘤模型,瘤体内注射Ad-LASP1-shRNA,定期测量肿瘤体积。结果NSCLC癌组织和癌旁组织中LASP1的mRNA表达量、蛋白表达水平差异均有统计学意义(P<0.001)。qPCR和Western blot表明Ad-LASP1-shRNA可有效抑制LASP1的表达。Ad-LASP1-shRNA感染A549细胞24 h后的A549迁移、侵袭能力显著降低,侵袭和迁移细胞数目显著低于对照组(P<0.05);接种5周后Ad-LASP1-shRNA组肿瘤体积为(334.5±32.6)mm 3明显小于PBS组(833.9±61.3)mm^(3),Ad-LASP1-shRNA能显著抑制移植瘤的生长。结论Ad-LASP1-shRNA对NSCLC有显著的抑瘤效果,值得进一步研究。

Objective To investigate the antitumor effect of interfering adenovirus targeting LIM and SH3 protein 1(LASP1)gene(Ad-LASP1-shRNA)on non-small cell lung cancer(NSCLC).Methods Real-time fluorescence quantitative polymerase chain reaction(qPCR)and Western blot were used to detect the levels of LASP1 mRNA and protein in NSCLC tissue.Then,A549 cells were transfected with Ad-LASP1-shRNA for 48 h.The expression of LASP1 mRNA and protein in the transfected A549 cells were analyzed by qPCR and Western blot.The wound healing and Transwell assay were used to detect the migration and invasion ability.A xenograft model was established using A549 cells.Then,Ad-LASP1-shRNA was intra-tumorally injected and tumor volume was measured regularly.Results NSCLC tissue and normal adjacent tissue showed statistical different relative amounts of LASP1 mRNA and protein(P<0.001).According to qPCR and Western blot,Ad-LASP1-shRNA effectively inhibited the expression of LASP1.After transfection with Ad-LASP1-shRNA for 24 h,the A549 cells presented remarkable increases in the abilities of migration and invasion,and the number of invasive and migratory cells,compared with the control group(P<0.05).After implantation for five weeks,the Ad-LASP1-shRNA group showed a tumor volume of(334.5±32.6)mm 3,which were significantly smaller than that of the PBS group(833.9±61.3)mm^(3)(P<0.001).Ad-LASP1-shRNA was able to significantly inhibit the growth of xenograft tumor.Conclusions Ad-LASP1-shRNA shows obvious anti-tumor effects and underlying mechanisms needed further investigation.