血管内皮生长因子165基因对人胃癌细胞在体外生长及其受体表达的影响
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关云艳, 欧希龙, 孙为豪, 颜 芳, 产松苗, 杨 柳
腺病毒;血管内皮生长因子;胃癌;增殖;VEGFR-2,关云艳,欧希龙,颜芳,杨柳,孙为豪,产松苗,关云艳,通讯作者:,Effectofvascularendothelialgrowthfactorgeneonitsr
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关云艳, 欧希龙, 颜芳, 杨柳, 东南大学临床医学院附属中大医院消化科 江苏省南京市 210009
孙为豪, 南京医科大学第一附属医院老年病科 江苏省南京市 210029
产松苗, 上海梅山医院消化科 江苏省南京市 210039
关云艳, 东南大学临床医学院研究生, 主治医师, 主要从事消化系统肿瘤的研究.
通讯作者: 欧希龙, 210009, 江苏省南京市, 东南大学临床医学院附属中大医院消化科. ouxilong@126.com
电话: 025-83272034
收稿日期: 2006-12-04 接受日期: 2006-12-31
Effect of vascular endothelial growth factor gene on its receptor-2 expression and proliferation of human gastric carcinoma cell in vitro
Yun-Yan Guan, Xi-Long Ou, Wei-Hao Sun, Fang Yan, Song-Miao Chan, Liu Yang
Yun-Yan Guan, Xi-Long Ou, Fang Yan, Liu Yang,Department of Gastroenterology, Zhongda Hospital Affiliated to Clinical Medical College of Southeast University, Nanjing 210009, Jiangsu Province, China
Wei-Hao Sun,Department of Geratology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China
Song-Miao Chan,Department of Gastroenterology, Shanghai Meishan Hospital, Nanjing 210039, Jiangsu Province, China
Correspondence to:Xi-Long Ou, Department of Gastroenterology, Zhongda Hospital Affiliated to Clinical Medical College of Southeast University, Nanjing 210009, Jiangsu Province, China. ouxilong@126.com
Received:2006-12-04 Accepted:2006-12-31
Abstract
AIM: To amplify and purify the recombinant adenovirus vector carrying vascular endothelial growth factor 165 (Ad-VEGF165), and to investi-gate its effect of on the expression of VEGF receptor-2 (VEGFR-2) and the proliferation of human gastric adenocarcinoma cell line BGC-823 in vitro.
METHODS: The adenoviruses were packaged and amplified in human embryonic kidney 293 cells. Then human gastric adenocarcinoma cell line BGC-823 was transfected with the resultant adenoviruses. The proliferation and expression of VEGFR-2 were measured by MTT assay and immunocytochemistry, respectively.
RESULTS: The recombinant adenovirus vector carrying VEGF165 or GFP (control virus) was amplified and purified successfully. The viral titer of VEGF165 or GFP was 3.2 × 1013 and 2.0 × 1013 pfu/L, respectively. The transfection rate almost reached 100% in BGC-823 cells when the multiplicity of infection (MOI) was 20. The proliferation of BGC-823 cells (shown by absorbency) transfected with Ad-VEGF165 was significantly higher than that of the cells transfected with Ad-GFP (24 h: 0.960 ± 0.01 vs 0.737 ± 0.01, P < 0.01; 48 h: 1.321 ± 0.03 vs 0.981 ± 0.02, P < 0.01; 72 h: 1.663 ± 0.03 vs 1.207 ± 0.01, P < 0.01) or the controlled cells (24 h: 0.960 ± 0.01 vs 0.724 ± 0.03, P < 0.01; 48 h: 1.321 ± 0.03 vs 0.968 ± 0.01, P < 0.01; 72 h: 1.663 ± 0.03 vs 1.185 ± 0.02, P < 0.01). Moreover, the expression of VEGFR-2 was expressed was markedly higher in BGC-823 cells transfected with Ad-VEGF165 than that in Ad-GFP-transfected cells (62.5% vs 37 ......
关云艳, 欧希龙, 颜芳, 杨柳, 东南大学临床医学院附属中大医院消化科 江苏省南京市 210009
孙为豪, 南京医科大学第一附属医院老年病科 江苏省南京市 210029
产松苗, 上海梅山医院消化科 江苏省南京市 210039
关云艳, 东南大学临床医学院研究生, 主治医师, 主要从事消化系统肿瘤的研究.
通讯作者: 欧希龙, 210009, 江苏省南京市, 东南大学临床医学院附属中大医院消化科. ouxilong@126.com
电话: 025-83272034
收稿日期: 2006-12-04 接受日期: 2006-12-31
Effect of vascular endothelial growth factor gene on its receptor-2 expression and proliferation of human gastric carcinoma cell in vitro
Yun-Yan Guan, Xi-Long Ou, Wei-Hao Sun, Fang Yan, Song-Miao Chan, Liu Yang
Yun-Yan Guan, Xi-Long Ou, Fang Yan, Liu Yang,Department of Gastroenterology, Zhongda Hospital Affiliated to Clinical Medical College of Southeast University, Nanjing 210009, Jiangsu Province, China
Wei-Hao Sun,Department of Geratology, the First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, Jiangsu Province, China
Song-Miao Chan,Department of Gastroenterology, Shanghai Meishan Hospital, Nanjing 210039, Jiangsu Province, China
Correspondence to:Xi-Long Ou, Department of Gastroenterology, Zhongda Hospital Affiliated to Clinical Medical College of Southeast University, Nanjing 210009, Jiangsu Province, China. ouxilong@126.com
Received:2006-12-04 Accepted:2006-12-31
Abstract
AIM: To amplify and purify the recombinant adenovirus vector carrying vascular endothelial growth factor 165 (Ad-VEGF165), and to investi-gate its effect of on the expression of VEGF receptor-2 (VEGFR-2) and the proliferation of human gastric adenocarcinoma cell line BGC-823 in vitro.
METHODS: The adenoviruses were packaged and amplified in human embryonic kidney 293 cells. Then human gastric adenocarcinoma cell line BGC-823 was transfected with the resultant adenoviruses. The proliferation and expression of VEGFR-2 were measured by MTT assay and immunocytochemistry, respectively.
RESULTS: The recombinant adenovirus vector carrying VEGF165 or GFP (control virus) was amplified and purified successfully. The viral titer of VEGF165 or GFP was 3.2 × 1013 and 2.0 × 1013 pfu/L, respectively. The transfection rate almost reached 100% in BGC-823 cells when the multiplicity of infection (MOI) was 20. The proliferation of BGC-823 cells (shown by absorbency) transfected with Ad-VEGF165 was significantly higher than that of the cells transfected with Ad-GFP (24 h: 0.960 ± 0.01 vs 0.737 ± 0.01, P < 0.01; 48 h: 1.321 ± 0.03 vs 0.981 ± 0.02, P < 0.01; 72 h: 1.663 ± 0.03 vs 1.207 ± 0.01, P < 0.01) or the controlled cells (24 h: 0.960 ± 0.01 vs 0.724 ± 0.03, P < 0.01; 48 h: 1.321 ± 0.03 vs 0.968 ± 0.01, P < 0.01; 72 h: 1.663 ± 0.03 vs 1.185 ± 0.02, P < 0.01). Moreover, the expression of VEGFR-2 was expressed was markedly higher in BGC-823 cells transfected with Ad-VEGF165 than that in Ad-GFP-transfected cells (62.5% vs 37 ......
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