SiRNA, shRNA sequence

HNSCC cell lines PCI-4A/4B and PCI-37A/37B were supplied by the University of Pittsburgh Cancer Institute (USA). These cell lines were cultured at 37 °C in a humidified atmosphere of 5% CO₂ in the medium of Dulbecco's modified Eagle's medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA), 100U/ml penicillin G and 100U/ml streptomycin. The RNA oligonucleotide miR-92b mimics, inhibitor, SP1 siRNA, and negative control were synthesized by Genepharma (2603, Genepharma, Shanghai, China). MiR-92b inhibitor and negative control plasmid were purchased from GeneChem (PMDE249003218, GeneChem, Shanghai, China). Transfection agent Lipofectamine 2000 (11668500, Invitrogen) was used according to the manufacturer’s protocol. Cells were harvested 24 hours after transfection for cell migration, invasion, proliferation and wound healing assay, 48 hours after for RNA isolation and 72 hours after for western blot analysis.This is the sequences of siRNA and shRNA used in this study.头颈部鳞状细胞癌（HNSCC）细胞系PCI-4A/4B与PCI-37A/37B由美国匹兹堡大学癌症研究所提供。 上述细胞系于37 ℃、含5% CO₂的饱和湿度培养环境中培养，培养基采用达尔伯克改良伊格尔培养基（Dulbecco's modified Eagle's medium, DMEM；Invitrogen，美国加利福尼亚州卡尔斯巴德），并添加10%胎牛血清（FBS；Gibco，美国加利福尼亚州卡尔斯巴德）、100 U/ml青霉素G及100 U/ml链霉素。 本研究所用的RNA寡核苷酸miR-92b模拟物、抑制剂、SP1小干扰RNA（SP1 siRNA）及阴性对照均由吉玛基因（Genepharma，中国上海，货号2603）合成。 miR-92b抑制剂与阴性对照质粒购自吉凯基因（GeneChem，中国上海，货号PMDE249003218）。 转染试剂脂质体2000（Lipofectamine 2000，货号11668500，Invitrogen）依照制造商说明书使用。 转染后24小时收集细胞，用于细胞迁移、侵袭、增殖及划痕愈合实验；转染后48小时收集细胞用于RNA提取；转染后72小时收集细胞用于蛋白质印迹（western blot）分析。 本研究所用小干扰RNA（siRNA）与短发夹RNA（shRNA）序列如下。