摘要
目的利用RNA干扰技术沉默人胃腺癌细胞株BGC-823中血管生成因子1(Ang-1)的表达,观察其对肿瘤侵袭性的抑制效果。方法设计靶向Ang-1的siRNA片段并转染人胃腺癌细胞株BGC-823;RT-PCR方法检测Ang-1的mRNA水平;Western Blot和细胞免疫荧光方法检测整合素β1、CD44V6和Ang-1的蛋白表达量和细胞定位;细胞黏附试验检测转染前后细胞黏附能力;Matrigel胶及Transwell双室培养体系检测癌细胞侵袭能力。结果 RT-PCR结果显示所设计的siRNA片段可有效沉默Ang-1的mRNA表达;整合素β1、CD44V6和Ang-1蛋白主要定位于肿瘤细胞胞浆和胞膜,其表达水平较转染siRNA片段前均有不同程度降低;细胞黏附能力和侵袭能力均明显降低(P<0.01)。结论靶向Ang-1的siRNA技术可有效降低人胃腺癌细胞株BGC-823的侵袭能力,为胃癌基因治疗提供了新的思路。
Objective To knock down angiopoietin-1 expression in human gastric cancer cell line BGC-823 and to observe its effect of reversing tumor invasion. Methods siRNA sequence fragments was designed to target angiopoietin-1 and transferred into human gastric cancer cell line BGC-823. RT-PCR was used to assess the transcription level of angio-poietin-1 mRNA, then western blot and immunofluorescence were used to examine the expression level of three invasion-as-sociated proteins include integrinβ1, CD44V6 and Ang-1. Cell adhesion ability was evaluated by cell adhesion assay and cell invation was determined by matrigel and transwell plastic dual-chamber culture system. Results Ang-1 mRNA was knocked down by siRNA showed by RT-PCR. The expression of integrinβ1, CD44V6 and Ang-1 were significantly lower than control group（P〈0.05）, so did the cellular adhesion and invasion abilities（P〈0.05）. Conclusion Knocking down angiopoietin-1 by siRNA can reverse invasion of human gastric cancer cell line BGC-823 and may provide new ideas and reference for gene therapy of gastric cancer in the future.
出处
《天津医药》
CAS
北大核心
2014年第8期752-754,I0001,共4页
Tianjin Medical Journal
