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).