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. 2016 Nov 16;7(52):86313–86325. doi: 10.18632/oncotarget.13388

Figure 4. Phenformin inhibits the development of selumetinib resistance in osimertinib resistant cell lines.

Figure 4

PC9-AZDR1 (A) and NCI-H1975-AZDR1 (B) cells were plated at low density, treated with the indicated compounds and scored for resistance as described in the Materials and Methods. (C) NCI-H1975-AZDR1 and NCI-H1975-AZDR4 cells were treated as follows, lysed and subjected to Western blotting; lane 1. 160 nM osimertinib; lane 2. Osimertinib/300 nM selumetinib; lane 3. Osimertinib/100 μM phenformin; lane 4. Osimertinib/ selumetinib/phenformin. (D) Indicated cell lines were treated for 48 h with osimertinib (160 nM) + selumetinib (300 nM), osimertinib + phenformin (30 μM) or osimertinib + selumetinib + phenformin and subjected to a caspase 3/7 assay as described in the Materials and Methods. Values shown are means relative to osimertinib-only control +/− SEM (*p < 0.05; n = 3). (E) Cells were treated for 24 h with osimertinib (Osi; 160 nM), selumetinib (Sel; 300 nM) or both (Osi/Sel), and conditioned media was analysed for lactate concentration. Values shown are means relative to osimertinib-only control +/− SEM (*p < 0.05; n = 3).