VEGF165 antisense RNA suppresses oncogenic properties of human esophageal squamous cell carcinoma.
OBJECTIVE
To investigate the effect of antisense RNA to vascular endothelial growth factor165 (VEGF165) on human esophageal squamous cell carcinoma cell line EC109 and the feasibility of gene therapy for esophageal carcinoma.


METHODS
By using subclone technique, the full length of VEGF165 amino acid cDNA, which was cut from pGEM-3Zf+,was cloned inversely into the eukaryotic expression vector pCEP4. The recombinant plasmid pCEP-AVEGF165 was transfected into EC109 cell with lipofectamine. After a stable transfection, dot blot, enzyme-linked immunosorbent assay (ELISA),laser confocal imaging system analysis, transmission electron microscopy and flow cytometry were performed to determine the biological characteristics of EC109 cell line before and after transfection in vitro and whether there was a reversion in the tumorigenic properties of the EC109 cell in vitro.


RESULTS
The eukaryotic expression vector pCEP-AVEGF165 was successfully constructed and transfected into EC109 cells. The expression of VEGF165 was significantly decreased in the transfected cells while the biological characteristics of the cells were not influenced by the expression of antisense gene. The tumorigenic and angiogenic capabilities were greatly reduced in nude mice, as demonstrated by reduced tumor end volume (820+/-112.5)mm(3) vs (7930+/-1035)mm(3) and (7850+/-950)mm(3), P<0.01) and microvessel density (average number: (8.5+/-1.2)mm(-2) vs (44.3+/-9.4)mm(-2) and (46.4+/-12.6)mm(-2), P<0.01) in comparison between experimental groups empty vector transfected group and control group.


CONCLUSIONS
The angiogenesis and tumorigenicity of human esophageal squamous cell carcinoma were effectively inhibited by VEGF165 antisense RNA. Antisense RNA to VEGF165 can potentially be used as an adjuvant therapy for solid tumors.
Gu
Zhong-Ping
ZP
Department of Thoracic Surgery, Fourth Military Medical University, Xi'an 710038, Shaanxi Province,China. zhongpg@pub.xaonline.com
Wang
Yun-Jie
YJ
Li
Jin-Ge
JG
Zhou
Yong-An
YA
D000818:Animals; D015197:Carcinogenicity Tests; D002294:Carcinoma, Squamous Cell / Q000503:physiopathology / Q000628:therapy*; D002453:Cell Cycle / Q000235:genetics; D003001:Cloning, Molecular; D016228:Endothelial Growth Factors / Q000235:genetics*; D004938:Esophageal Neoplasms / Q000503:physiopathology / Q000628:therapy*; D015972:Gene Expression Regulation, Neoplastic; D015316:Genetic Therapy; D006801:Humans; D008222:Lymphokines / Q000235:genetics*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008854:Microscopy, Electron; D009389:Neovascularization, Pathologic / Q000517:prevention & control / Q000628:therapy; D010957:Plasmids; D016372:RNA, Antisense / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000502:physiology / Q000648:ultrastructure; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D016228:Endothelial Growth Factors; D008222:Lymphokines; D016372:RNA, Antisense; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors