Gdc 0941

The PI3K inhibitors AZD8835, AMG319, HS173, and AS605240 were purchased from Selleck (Houston, TX, USA). The PI3K inhibitors GDC0941, LY294002, and BEZ235 and the Akt inhibitor afuresertib were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Each drug was dissolved with dimethylsulfoxide (DMSO) to a storage solution of 100 mM. We diluted 100 mM of each drug with DMSO to a concentration of 10 mM of drug. BmE cells were seeded in 96-well plates 24 h before treatment with drugs. We added 10 mM of each drug to the cell culture medium at a dilution ratio of 1:1000 (0.1% of total volume) to prepare 10 µM of the drug. BmE cells were incubated with 10 µM of the drugs for 72 h. BmE cells were also incubated with 0 µM, 2.5 µM, 5 µM, 10 µM, 20 µM, 40 µM, 80 µM, and 100 µM of AZD8835 for 72 h. Each treatment with drugs and DMSO control comprised three replicates. Cell viability was measured using the CellTiter 96^® AQueous One Solution Cell Proliferation Assay (MTS) (Promega, Madison, WI, USA) [22 (link)].

Breast cancer cell lines BT-474, MCF7, Hs-578-T, MDA-MB-231, MDA-MB-453, and MDA-MB-468 (Cobioer, Nanjing, China) were cultured in Dulbecco’s Modified Eagle Medium (DMEM, Sunncell, Wuhan, China) with 10% fetal bovine serum (FBS) and certain cell growth factor. BT-549, HCC1937, and T47D cell lines were cultured in Roswell Park Memorial Institute (RPMI)-1640 medium (Sunncell, Wuhan, China) with 10% or 20% FBS.
For the following function experiments, T47D, MCF7, MDA-MB-453, HCC1937, MDA-MB-231, and MDA-MB-468 cells were treated with 100 or 500 nM GDC-0941 (PI3K selective inhibitor, MedChemExpress, New Jersey, USA) or dimethyl sulfoxide (DMSO) for 24 h. In addition, MDA-MB-231 and HCC1937 cells were treated with 100 nM OSU-T315 (ILK inhibitor, SolelyBio, Beijing, China), tumor necrosis factor (TNF)-α, IgG, or TNF-α antibody (ab6671, Abcam, Cambridge, UK).
In addition, MCF7 and MDA-MB-453 cells were transfected with ILK, and MDA-MB-231 and HCC1937 cells were transfected with shILK.