敲低Abl相互作用蛋白1的表达抑制胃癌细胞NCI-N87的体外增殖和迁移

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.

Abstract：

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.

Abl interactor 1 knock-down inhibits the in vitro proliferation and migration of NCI-N87 gastric cancer cells

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.