Figure 6
From: Targeting Mcl-1 by a small molecule NSC260594 for triple-negative breast cancer therapy
The combination treatment of NSC260594 and everolimus acted synergistically to kill TNBCs. (A) NSC260594 and everolimus (Rapa) synergistically killed MDA-MB 231, MDA-MB157 and MDA-MB468 cells. (B) NSC260594 and everolimus (Rapa) acted synergistically to kill TU-BcX-4QATb2 3D organoids. PDX-Os were formed from the digested PDX tumor tissue, and then 2000 cells per well were plated in a low attachment 24 well plate. After TU-BcX-4QATb2 organoids were treated with DMSO or NSC260594 (10 μM), Everolimus (20 μM) and 10 μM NSC260594 + 20 μM Everolimus for 72 h, cells were stained with Calcein-AM (2 μM) and Ethidium homodimer (EthD)-III (5 μM) for 45 min using the PromoKine live/dead staining kit. Organoids were imaged with fluorescence microscopy. The 588 nM excitation channel was used to identify red, ‘dead’ cells, and the 420 nM excitation channel was used to visualize green, ‘live’ cells. (C) The number of live organoids per filed was quantified. (D) MDA-MB 157 cells were treated with DMSO (control), NSC260594, everolimus, and the combination of NSC260594 and everolimus. Expression of mTOR cascade proteins in cell lysates was analyzed by western blot assays. (E) The diagram of the potential molecular mechanisms underlying the synergistic effect of NSC260594 and everolimus in killing TNBC cells.
