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（Head and Neck Squamous Cell Carcinoma，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的湿润培养环境中培养，所用培养基为添加10%胎牛血清（FBS；Gibco，美国加利福尼亚州卡尔斯巴德）、100U/ml青霉素G及100U/ml链霉素的杜氏改良伊格尔培养基（Dulbecco's modified Eagle's medium，DMEM；Invitrogen，美国加利福尼亚州卡尔斯巴德）。本研究所需的RNA寡核苷酸包括miR-92b模拟物、抑制剂、SP1小干扰RNA（Small Interfering RNA，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Lipofectamine 2000（货号11668500，Invitrogen）按照厂商说明书操作。细胞于转染后24小时收获，用于细胞迁移、侵袭、增殖及划痕愈合实验；转染后48小时收获用于RNA提取；转染后72小时收获用于蛋白质印迹分析。本研究所用siRNA与短发卡RNA（short hairpin RNA，shRNA）序列如下。