摘要
目的 观察血管内皮生长因子1 6 5反义 RNA对人喉鳞癌细胞 Hep- 2的作用 ,探讨其治疗喉癌的可行性 .方法 采用亚克隆技术 ,构建并鉴定反义 VEGF1 6 5真核表达载体 ;重组质粒转染人喉鳞癌细胞 Hep- 2后 ,将其接种于裸鼠皮下 ,利用原位杂交、EL ISA、激光共聚焦、图像分析及微血管计数等方法 ,观察转染前后 Hep- 2细胞的生物学性状和致瘤性的改变 .结果 构建的 VEGF1 6 5反义真核表达载体在 Hep- 2细胞中获得表达 ,转染后的细胞 VEGF1 6 5的表达下降 70 % ,其生物学性状不受外源基因表达的影响 ,但其在裸鼠皮下的致瘤性和血管生成能力明显下降 ,实验组、空载体组和对照组肿瘤体积分别为 (736± 2 6 2 ) ,(74 4 0± 335 )和 (772 0± 35 0 ) mm3(P<0 .0 1) ,微血管密度分别为 (9.6± 5 .4 ) ,(45 .5± 8.4 )和 (48.4± 7.6 ) mm- 2 (P<0 .0 1) .结论 VEGF1 6 5反义 RNA能够显著减少喉癌细胞内 VEGF1 6 5的表达 ,具有抑制肿瘤生长和血管生成的作用 ,有望成为抗喉癌的新基因治疗手段 .
AIM To investigate the effect of vascular endothelial growth factor 165 (VEGF 165 ) antisense RNA on human laryngeal squamous cell carcinoma cell line Hep 2 and the feasibility of gene therapy for laryngeal carcinoma.METHODS The eukaryotic expression vector of antisense VEGF 165 was constructed and identified by using subclone technique. The recombinant plasmid pCEP AVEGF 165 was transfected into Hep 2 cell by using Lipofectamine 2000. After a stable transfection, in situ hybridization, enzyme linked immunosorbent assay (ELISA), laser confocal imaging system analysis, transmission electron microscopy and flow cytometry were performed to determine the biological characteristics of Hep 2 cell before and after transfection in vitro and whether there was a reversion in the tumorigenic properties of the Hep 2 cell in vivo . RESULTS The eukaryotic expression vector pCEP AVEGF 165 was successfully constructed and transfected into Hep 2 cell. The biological characteristics of the cells were not influenced by the expression of antisense gene. The tumorigenic and angiogenic capabilities were significantly reduced in nude mice, as demonstrated by reduced tumor volume [experimental group: (736±262) mm 3, empty vector transfected group: (7440±335) mm 3, control group: (7720±350) mm 3; P <0.01] and microvessel density in experimental group compared with empty vector transfected group and control group [(9.6±5.4), (45.5±8.4) and (48.4±7.6) mm -2 , P <0.01]. CONCLUSION The angiogenesis and tumorigenicity of human laryngeal squamous cell carcinoma are effectively inhibited by VEGF 165 antisense RNA. Antisense RNA therapy to VEGF 165 can potentially become a new therapeutic method for laryngeal carcinoma.
出处
《第四军医大学学报》
北大核心
2002年第19期1773-1775,共3页
Journal of the Fourth Military Medical University
关键词
血管内皮生长因子
喉癌
反义RNA
基因治疗
vascular endothelial growth factor
antisense RNA
laryngeal carcinoma
gene therapy
