CAJ | 학술논문

目的探讨敲低Abl相互作用蛋白1(Abl interactor 1,ABI-1)对胃癌NCI-N87细胞体外增殖和迁移能力的影响.方法利用脂质体和嘌呤霉素筛选稳定表达ABI-1短发夹RNA(short hairpin RNA,ShRNA)的NCI-N87模型细胞,用实时定量RT-PCR和Western blot鉴定ABI-1敲低的效果;用CCK-8试剂盒、细胞骨架染色和Transwell小室检测ABI-1敲低对细胞增殖、形态和骨架以及迁移的影响;用Western blot检测磷酸化AKT蛋白的表达.结果成功获得表达ABI-1-ShRNA的NCI-N87细胞模型.CCK-8法检测显示NCI-N87-Vector和NCI-N87细胞在36 h和48 h的增殖率之间相比差异均无统计学意义(t=0.400、0.489,P＞0.05),而NCI-N87-ABI-1-ShRNA在36 h和48 h这2个时间点的增殖率均低于NCI-N87细胞(t=2.85、4.166,P＜0.05),ABI-1敲低抑制NCI-N87细胞的增殖.细胞骨架染色显示ABI-1敲低能够改变90%NCI-N87细胞的形态和骨架结构.Transwell研究显示NCI-N87、NCI-N87-Vector和NCI-N87-ABI-1-ShRNA的细胞迁移数分别是66±8、65±8和30±4,前两者相比差异无统计学意义(t=0.269,P＞0.05),敲低组与NCI-N87相比差异有统计学意义(t=9.550,P＜0.05),ABI-1敲低抑制NCI-N87细胞的迁移.Western Blot显示ABI-1敲低抑制磷酸化AKT蛋白的表达.结论 ABI-1敲低可能通过PI3K/AKT通路抑制胃癌细胞NCI-N87的体外增殖和迁移.
목적 탐토고저Abl상호작용단백1(Abl interactor 1,ABI-1)대위암NCI-N87세포체외증식화천이능력적영향.방법 이용지질체화표령매소사선은정표체ABI-1단발협RNA(short hairpin RNA,ShRNA)적NCI-N87모형세포,용실시정량RT-PCR화Western blot감정ABI-1고저적효과;용CCK-8시제합、세포골가염색화Transwell소실검측ABI-1고저대세포증식、형태화골가이급천이적영향;용Western blot검측린산화AKT단백적표체.결과 성공획득표체ABI-1-ShRNA적NCI-N87세포모형.CCK-8법검측현시NCI-N87-Vector화NCI-N87세포재36 h화48 h적증식솔지간상비차이균무통계학의의(t=0.400、0.489,P＞0.05),이NCI-N87-ABI-1-ShRNA재36 h화48 h저2개시간점적증식솔균저우NCI-N87세포(t=2.85、4.166,P＜0.05),ABI-1고저억제NCI-N87세포적증식.세포골가염색현시ABI-1고저능구개변90%NCI-N87세포적형태화골가결구.Transwell연구현시NCI-N87、NCI-N87-Vector화NCI-N87-ABI-1-ShRNA적세포천이수분별시66±8、65±8화30±4,전량자상비차이무통계학의의(t=0.269,P＞0.05),고저조여NCI-N87상비차이유통계학의의(t=9.550,P＜0.05),ABI-1고저억제NCI-N87세포적천이.Western Blot현시ABI-1고저억제린산화AKT단백적표체.결론 ABI-1고저가능통과PI3K/AKT통로억제위암세포NCI-N87적체외증식화천이.
Objective To investigate the effects of ABI-1 gene knockdown upon the proliferation and migration of human gastric cancer cell NCI-N87 in vitro. Methods NCI-N87-ABI-I-ShRNA cell model was successfully constructed and validated by Real-time PCR and Western blot. The cellular morphous and skeleton, proliferative and migrative potents, and also AKT expression were compared between NCI-N87-ABI-1-ShRNA and its parents by immunofluorental staining, CCK-8 assay, transwell chamber and Western blotting. Results CCK-8 assay showed there was no significant difference in the proliferation rates at different time points between the NCI-N87-Vector and NCI-N87 cells while the proliferation rates at the time points of 36 and 48 hours of the NCI-N87-ABI-1-ShRNA were significantly lower than the NCI-N87( t =2. 85and 4. 166, P ＜ 0. 05 ). Transwell assay showed that migrated cell number were 66 ± 8, 65 ± 8 and 30 ± 4,respectively, and there was significant difference between the NCI-N87-ABI-1-ShRNA and NCI-N87 cells (t =9. 550,P ＜0. 05). Finally, ABI-1- knock-down altered the cellular morphoos and skeleton of 90％NCI-N87 cells and inhibited p-AKT expression. Conclusion ABI-1 inhibits proliferation and migration of NCI-N87 cells in vitro probably by PI3K/AKT pathway.