CAJ | 학술논문

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。
목적：이용RNA간우기술침묵인위선암세포주BGC-823중혈관생성인자1（Ang-1）적표체，관찰기대종류침습성적억제효과。방법설계파향Ang-1적siRNA편단병전염인위선암세포주BGC-823；RT-PCR방법검측Ang-1적mRNA수평；Western Blot화세포면역형광방법검측정합소β1、CD44V6화Ang-1적단백표체량화세포정위；세포점부시험검측전염전후세포점부능력；Matrigel효급Transwell쌍실배양체계검측암세포침습능력。결과 RT-PCR결과현시소설계적siRNA편단가유효침묵Ang-1적mRNA표체；정합소β1、CD44V6화Ang-1단백주요정위우종류세포포장화포막，기표체수평교전염siRNA편단전균유불동정도강저；세포점부능력화침습능력균명현강저（P<0.01）。결론파향Ang-1적siRNA기술가유효강저인위선암세포주BGC-823적침습능력，위위암기인치료제공료신적사로。
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.