Effects of exogenous antisense VEGF-C gene on growth of human gastric carcinoma cell line SGC-7901 in vitro

ZHU Peng; et al

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Effects of exogenous antisense VEGF-C gene on growth of human gastric carcinoma cell line SGC-7901 in vitro

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- Effects of exogenous antisense VEGF-C gene on growth of human gastric carcinoma cell line SGC-7901 in vitro
- Journal of Chongqing Medical University Vol. 34, Issue 5, Pages: 558-562(2009)
- 作者机构：
  
  重庆医科大学附属第二医院普外科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R735.2
- Published：2009，
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[1]朱鹏,张剑波,王继见.外源性反义血管内皮生长因子-C基因稳定转染对人胃癌细胞SGC-7901体外生长的影响[J].重庆医科大学学报,2009,34(05):558-562.

ZHU Peng, et al. Effects of exogenous antisense VEGF-C gene on growth of human gastric carcinoma cell line SGC-7901 in vitro. [J]. Journal of Chongqing Medical University 34(5):558-562(2009)
[1]朱鹏,张剑波,王继见.外源性反义血管内皮生长因子-C基因稳定转染对人胃癌细胞SGC-7901体外生长的影响[J].重庆医科大学学报,2009,34(05):558-562. DOI：

ZHU Peng, et al. Effects of exogenous antisense VEGF-C gene on growth of human gastric carcinoma cell line SGC-7901 in vitro. [J]. Journal of Chongqing Medical University 34(5):558-562(2009) DOI：

摘要

目的:观察以pCI-neo为载体的血管内皮生长因子-C（Vascular endothelial growth factor-C

VEGF-C）反义基因转染的人胃癌细胞SGC-7901中VEGF-C蛋白的表达

并研究其对人胃癌细胞体外生长的影响。方法:应用重组质粒pCI-neo-anti VEGF-C和pCI-neo空载体质粒

以脂质体转染法转染体外培养的人胃癌细胞株

以未转染组为对照

经G418筛选抗性克隆

扩大培养后

再用免疫组化和Western-blot分析VEGF-C基因及蛋白的表达

最后用MTT法分析其对细胞生长的抑制作用。结果:免疫组化和Western-blot均显示pCI-neo-anti VEGF-C转染组无VEGF-C mRNA及其蛋白的表达

MTT检测表明pCI-neo-anti VEGF-C转染组活细胞数较其它各组均低（P<0.05）。结论:pCI-neo-anti VEGF-C成功转染人胃癌细胞SGC-7901且可封闭VEGF-C mRNA

使已转染pCI-neo-anti VEGF-C的人胃癌细胞VEGF-C蛋白的表达减少

并能抑制胃癌细胞SGC-7901的体外生长

为进一步研究VEGF-C的生物学功能及胃癌的基因治疗研究创造了条件。

Abstract

Objective:To observe the expression of VEGF-C protein in human gastric cancer cells SGC-7901 those transfected by pCI-neo-antisense VEGF-C and investigate whether antisense VEGF-C gene could inhibit the growth of human gastric carcinoma cell line in vitro.Methods:By the liposome infection protocol

We transfected two different plasmids including pCI-neo and pCI-neo antisense VEGF-C into human gastric cancer cell line SGC-7901 cultured in vitro.Then those resistant clones were selected by G418 and expanded in DMEM culture medium.Then we used Immunohistochemical method and Western-blot to detect target genes and its expression.The cell viability was detected by MTT assay.We took the untransfected group as the control.Results:The immunohistochemical method and Western-blot showed that there were no expression of VEGF-C mRNA and its protein in the group that transfected by plasmid pCI-neo antisense VEGF-C.MTT assay showed that the group of transfected with antisense VEGF-C had a lower cell viability than other groups(P<0.05).Conclusion:The target genes were successfully transfected and the expression of VEGF-C mRNA and its protein in SGC-7901 cells could be inhibited by pCI-neo-antisense VEGF-C.The study also showed that antisense VEGF-C gene could suppress the growth of human gastric cells in vitro.These results created the condition for further study of the biologic functions of VEGF-C and gene therapy for gastria carcinoma.

关键词

人胃癌细胞株反义寡核苷酸类血管内皮生长因子-C基因转染

Keywords

cell line of gastric carcinomaAntisense oligonucleotidesVascular endothelial factor-c growth(VEGF-C)gene transfection

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