Transwell data

Migration and invasion assays were performed using Transwell chambers (3422, 24-well insert; 8-μm pore size; Corning Costar, NY, USA) coated with or without Matrigel (356234, BD Biosciences, NJ, USA). 4 × 10⁵HNSCC cells in DMED medium were placed into the upper chamber, and medium supplementary with 10 % FBS was added to the lower chamber. Then PCI-37A cells were incubated for 24 hours for migration and 48 hours for invasion, and PCI-37A cells were incubated for 8 hours for migration and 24 hours for invasion. Then the migratory or invasive cells were fixed by 4% paraformaldehyde, stained with 0.1% crystal violet, and counted in three random fields under 200x magnification. Transwell migration assay and Martrigel invasion assay showed that an ectopic increased level of miR-92b could raise the migration and invasion ability of PCI-37A cells, while a reduced level of miR-92b led to a significant decrease in the number of migrating or invasive cells.After knocked down with SP1 expression, PCI-37A/B cells showed a significant decrease in migration and invasion.本研究采用包被或未包被基质胶（Matrigel，货号356234，BD Biosciences，美国新泽西州）的Transwell小室（Transwell，货号3422，24孔插入式，孔径8 μm，Corning Costar，美国纽约州）进行细胞迁移与侵袭实验。将4×10^5头颈部鳞状细胞癌（HNSCC，Head and Neck Squamous Cell Carcinoma）细胞置于含DMED培养基的上室，下室加入添加10%胎牛血清（FBS）的培养基。其中PCI-37A细胞的迁移实验孵育时长为24 h、侵袭实验为48 h，另一组PCI-37A细胞的迁移实验孵育时长为8 h、侵袭实验为24 h。随后用4%多聚甲醛固定已迁移或侵袭至小室膜下方的细胞，以0.1%结晶紫染色，并在200倍放大视野下随机选取3个视野进行细胞计数。Transwell迁移实验与基质胶侵袭实验结果显示，异位升高miR-92b的表达水平可增强PCI-37A细胞的迁移与侵袭能力，而下调miR-92b的表达则会显著减少迁移及侵袭的细胞数量。敲低特异性蛋白1（SP1）的表达后，PCI-37A/B细胞的迁移与侵袭能力均出现显著下降。