[Experimental studies on inhibition of malignant melanoma growth by gene transfer of adenovirus-mediated averse vascular endothelial growth factor165].
OBJECTIVE
To investigate the effect of Adenovirus-mediated averse vascular endothelial growth factor165(Ad-aVEGF165)on the growth of human melanoma cells(A375) in vivo and in vitro.


METHODS
In vitro, the 100 multiplicity of infection of Adenovirus- mediated green fluorescent protein(Ad-GFP)and Ad-aVEGF165 were transfected into human endothelium cell of vessel304(ECV304) and A375. ECV304 cells were divided into 3 groups: A375 group, Ad-GFP group and Ad-aVEGF165 group. A375 cells were also divided into 3 groups: 1640 group, Ad-GFP group and Ad-aVEGF165 group. Their effects were analyzed by proliferation assay, cell cycle, and VEGF expression. In vivo, A375 cells were injected into the axilla of the nude mouse. When the tumor formed, they were transplanted into another 15 mice. After treatment, the tumor was excised for naked eye observation, HE observation and micro-vascular density (MVD) counting.


RESULTS
The cell supernatant fluid of A375 group and Ad-GFP group could stimulate ECV304 cell growth, but that of Ad-aVEGF165 group could inhibit the growth of ECV304 cell. All the A375 cells in 3 groups had the proliferation trend, showing no statistically significant difference (P>0.05). ECV304 cell proliferation index (PI) in Ad-aVEGF165 group reduced (P<0.05). There was no statistically significant difference (P>0.05) in the PI of A375 cell. The A375 cell integral optical densities were 234.41 +/- 13.8 in 1640 group, 222.73 +/- 3.67 in Ad-GFP group and 180.84 +/- 6.34 in Ad-aVEGF165 group. The tumor volume in Ad-aVEGF165 group was smaller than that in Ad-GFP group and PBS group at 2 weeks after operation, the trend became much obvious with the time delay. Ad-aVEGF165 brought to much tissue necrosis under HE stain. The MVD of PBS group, Ad-GFP group and Ad-aVEGF165 group were 65 +/- 10/view, 52 +/- 11/view and 30 +/- 6/ view, respectively.


CONCLUSIONS
In Vitro, Ad-VEGF165 gene could inhibited ECV304 cells' growth by weakening VEGF expression of A375 cells. In vivo, Ad-aVEGF165 could inhibit the growth of human melanoma from blocking micro-vascular.
Cui
Zhengjun
Z
Department of Plastic and Burn Surgery, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R. China.
Cen
Ying
Y
Zhang
Jie
J
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D045744:Cell Line, Tumor; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D008545:Melanoma / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A