论文部分内容阅读
目的 研究人血管生成素 1 (Ang1 )基因转染人胃癌细胞系SGC790 1后 ,对肿瘤细胞生物学特性以及对小鼠体内致瘤力和血管生成的影响 ,探讨其在胃癌发生及血管生成中的作用。方法构建重组正、反义Ang1真核表达载体 ,采用脂质体转染法 ,将重组载体转染人胃癌细胞系SGC790 1 ,分别得到正、反义载体稳定转染的细胞株 7Ang1 +和 7Ang1 - ,并以半定量PCR及Westernblot方法鉴定转染效果。以MTT比色试验绘制生长曲线 ;以流式细胞仪检测细胞周期分布。通过裸鼠成瘤实验 ,比较转染前后小鼠体内成瘤能力的差别 ,并检测肿瘤组织微血管密度 (MVD) ,判定血管生成程度。结果 7Ang1 -组细胞Ang1在蛋白及mRNA水平的表达均下调。体内成瘤实验结果显示 ,空载体对照组、7Ang1 +组和 7Ang1 -组瘤体的平均重量 ( x±s)分别为 6 2 4 .0 0± 77.78,6 5 2 .6 7± 1 32 .0 7和2 93.0 0± 95 .5 4 ,7Ang1 -组的细胞成瘤性显著低于 7Ang1 +组和空载体对照组 (P <0 .0 1 )。空载体对照组、7Ang1 +组和 7Ang1 -组肿瘤组织的MVD( x±s)分别为 8.4 4± 1 .33,8.78± 1 .92和 6 .0 0± 1 .73,7Ang1 -组细胞的血管生成显著减少 (P <0 .0 1 )。结论 Ang1在胃癌形成和发展中可能通过诱生血管起促进作用 ,通过反义技术可部分阻断这种作用。
Objective To investigate the biological characteristics of tumor cells and their effect on the tumorigenicity and angiogenesis in vivo of human gastric cancer cell line SGC790 1 after transfection of human angiopoietin 1 (Ang1) gene in gastric cancer and angiogenesis Role. Methods The recombinant sense and antisense Ang1 eukaryotic expression vector was constructed and transfected into human gastric cancer cell line SGC7901 by lipofection. The positive cell lines 7Ang1 + and 7Ang1 -, and semi-quantitative PCR and Western blotting method to identify transfection. The growth curve was drawn by MTT colorimetric assay. The cell cycle distribution was detected by flow cytometry. The nude mouse tumorigenicity experiment was performed to compare the tumorigenic ability of the mice before and after transfection. The microvessel density (MVD) of the tumor tissue was detected to determine the degree of angiogenesis. Results The expression of Ang1 in 7Ang1 - cells was down - regulated at both protein and mRNA levels. The results of in vivo tumorigenicity showed that the mean weight (x ± s) of the control group, 7Ang1 + group and 7Ang1 - group were 624.0 ± 77.78 and 65.26 ± 1 32, respectively. The tumorigenicity of 7Ang1 - group was significantly lower than that of 7Ang1 + group and empty vector control group (P <0.01). The MVD (x ± s) of the control group, 7Ang1 + group and 7Ang1 - group were 8.44 ± 1.33, 8.78 ± 1.92 and 6.00 ± 1.73, respectively Angiogenesis was significantly reduced (P <0.01). Conclusion Ang1 may contribute to the formation and development of gastric cancer through the induction of blood vessels. This effect can be partially blocked by antisense technology.