Drugs and reagents
Icariin (purity >98%, as measured by high-performance liquid chromatograph analysis) was purchased from Energy Chemical (Shanghai, China) and dissolved at a concentration of 20 mM in 100% DMSO as a stock solution, stored at -20ºC, and diluted with medium before use. The medium containing 0.1% DMSO served as the control. 3-(4,5-dimethyl-2-thiazyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) Rh123, DCFH-DA, and N-acetyl cysteine (NAC) were bought from Sigma Chemical Co. (St. Louis, MO, USA). Antibodies against Bcl-2, Bax, cleaved caspase3, SIRT6, HSK9 Ac, H3, p-IκBα, NF-κB p65, Fibrillarin, E-cadherin, N-cadherin, MMP-2, and β-actin were purchased from Cell Signaling Technology (Beverly, MA, USA). The inhibitor of SIRT6 (oss-128167) was purchased from Selleck Chemicals (Houston, TX, USA). The anti-Ki-67 mouse monoclonal antibody was brought from Merck Millipore (Billerica, MA, USA). The Annexin V-FITC/PI Apoptosis Detection Kit was obtained from KeyGen Biotech (Nanjing, China).