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目的探讨促血管生成素1(Ang-1)、血管内皮生长因子165(VEGF-165)对人胃癌细胞中整合素β1的影响,初步研究其对肿瘤黏附的影响及可能的作用机制。方法利用腺病毒作为载体将已构建成功的含Ad-Ang-1+Ad-VEGF-165/2基因(A组)、Ad-Ang-1基因(B组)和Ad-VEGF基因(C组)的重组腺病毒瞬时转染人胃癌细胞株BGC-823,转染Ad-GFP作为阴性对照(D组),未转染的正常细胞作为空白对照(E组),通过细胞黏附法检测转染前后细胞与细胞外基质(ECM)黏附能力的改变,再分别使用半定量逆转录-聚合酶链反应(RT-PCR)和Western blot方法对三组细胞中整合素β1的mRNA和蛋白表达水平进行分析。结果细胞黏附实验显示转染Ad-Ang-1、Ad-VEGF-165、Ad-Ang-1+Ad-VEGF-165/2后,细胞与细胞外基底之间的黏附力明显增强(P<0.05),B组与C组无区别(P>0.05),A组的黏附率明显高于B组和C组(P<0.05);RT-PCR、Western blot结果显示整合素β1mRNA和蛋白水平在A组、B组、C组中的表达要明显高于D组及E组(P<0.05),其中在A组的表达最强。结论转染Ad-Ang-1、Ad-VEGF-165和Ad-Ang-1+Ad-VEGF-165/2能够提高人胃癌细胞BGC-823中整合素β1mRNA、蛋白的表达,从而促进肿瘤细胞的黏附和转移,A组的作用强于B组和C组。
Objective To investigate the effect of Ang-1 and VEGF-165 on integrin β1 in human gastric cancer cells, and to study its effect on tumor adhesion and its possible mechanism. Methods Ad-Ang-1 + Ad-VEGF-165/2 gene (group A), Ad-Ang-1 gene (group B) and Ad-VEGF gene (group C) were successfully constructed using adenovirus as a vector. The recombinant adenovirus was transiently transfected into human gastric cancer cell line BGC-823, transfected with Ad-GFP as a negative control (group D) and untransfected normal cells as a blank control (group E) Cells and extracellular matrix (ECM). The mRNA and protein expression of integrin β1 in three groups of cells were analyzed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively . Results The results of cell adhesion assay showed that the adhesion between the cells and the extracellular matrix was significantly increased after Ad-Ang-1, Ad-VEGF-165 and Ad-Ang-1 + Ad-VEGF- ), There was no difference between group B and group C (P> 0.05). The adhesion rate of group A was significantly higher than that of group B and group C (P <0.05). RT-PCR and Western blot showed that the integrin β1 mRNA and protein levels were in A The expression in group B, group C was significantly higher than that in group D and group E (P <0.05), and the expression in group A was the strongest. Conclusion Transfection of Ad-Ang-1, Ad-VEGF-165 and Ad-Ang-1 + Ad-VEGF-165/2 can enhance the expression of integrin β1 mRNA and protein in human gastric cancer cell line BGC-823 Adhesion and metastasis, the role of group A stronger than the B group and C group.