CAJ | 학술논문

目的探讨磷脂酰肌醇蛋白聚糖3( GPC3)在肝细胞癌(HCC)发生发展中的作用机制.方法设计靶向GPC3的siRNA,转染Huh7细胞后实时荧光定量聚合酶链反应(Real-time PCR)检测其对GPC3表达的抑制效果；选用抑制效果明显的小干扰RNA(siRNA)下调GPC3表达后,Western blot法检测GPC3的蛋白表达水平,并用流式细胞仪、荧光显微镜观察及Transwell法分别对Huh7细胞的周期、增殖和侵袭进行检测.结果成功筛选出1条抑制效率达到74.87％的特异靶向GPC3的siRNA,可以显著抑制GPC3的转录翻译；同时,可见Huh7细胞的增殖和侵袭受到抑制,但并不影响细胞周期.结论 GPC3的表达有利于HCC的增殖侵袭,其表达降低不利于HCC的发生发展.
목적 탐토린지선기순단백취당3( GPC3)재간세포암(HCC)발생발전중적작용궤제.방법 설계파향GPC3적siRNA,전염Huh7세포후실시형광정량취합매련반응(Real-time PCR)검측기대GPC3표체적억제효과；선용억제효과명현적소간우RNA(siRNA)하조GPC3표체후,Western blot법검측GPC3적단백표체수평,병용류식세포의、형광현미경관찰급Transwell법분별대Huh7세포적주기、증식화침습진행검측.결과 성공사선출1조억제효솔체도74.87％적특이파향GPC3적siRNA,가이현저억제GPC3적전록번역；동시,가견Huh7세포적증식화침습수도억제,단병불영향세포주기.결론 GPC3적표체유리우HCC적증식침습,기표체강저불리우HCC적발생발전.
Objective To study the action mechanism of Glypican-3 ( GPC3 ) in the development of hepatocellular carcinoma (HCC).Methods siRNA targeting GPC3 was designed and transfected into Huh7 cells,and the inhibitory effects on the expression of GPC3 was tested by using real-time polymerase chain reaction (PCR).The siRNA with obvious inhibitory effect was selected to down-regulate the GPC3 expression,and then the expression of GPC3 protein was detected by usig Wesnter blotting.The cell cycle,proliferation and invasion of Huh7 cells were assayed by using flow cytometry,fluorescence microscopy and Transwell,respectively.Results We successfully screened out a siRNA specifically targeting GPC3,its inhibition efficiency was 74.87％ and the siRNA could significantly inhibit the transcription and translation of GPC3.The knock-down of GPC3 could inhibit the proliferation and invasion of Huh7 cells,but had no effects on the cell cycle.Conclusion GPC3 expression facilitates the proliferation and invasion of HCC.