靶向Ang-1的siRNA片段逆转人胃腺癌细胞株BGC-823侵袭表型的研究

【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 RTPCR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。

Study of Reversing Invasion of Human Gastric Cancer Cell Line BGC-823by Targeting Angiopoietin-1Using siRNA

Objective To knock down the expression of angiopoietin-1 in human gastric cancer cell line BGC-823, and observe the treatment effect of reversing tumor invasion phenotype. Methods To design the siRNA sequence fragments targeting angiopoietin-1 and transfer it into human gastric cancer cell line BGC-823. RT-PCR method was used to explore the expression of angiopoietin-1 mRNA, western blot and immunofluorescence methods were used to test the expression of three invasion-associated proteins, including integrin β1, CD44V6 and Ang-1. Cell adhesion assay was employed to detect the cellular adhesion ability, matrigel and transwell plastic dual-chamber culture system for detection of cancer cell invasion. Results The siRNA sequence fragments can knock down Ang-1 mRNA level through RT-PCR results. The expression of integrin β1, CD44V6 and Ang-1 were lower significantly than control group（P<0.05）, as so as the cellular adhesion and invasion abilities（P<0.05）. Conclusion The siRNA technique targeting angiopoietin-1 can reverse the invasion phenotype of human gastric cancer cell line BGC-823, and may provide new ideas and reference for future gene therapy for gastric cancer.