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. 2022 Mar 15;82(6):1013–1024. doi: 10.1158/0008-5472.CAN-21-1707

Figure 5.

Figure 5. DAU and BAY-1895344 combination suppresses tumor growth 3D assays and in vivo. A, HCT116, RKO, and ARPE-19-MYC (DOX+) cells grown on Matrigel were treated with 5 μmol/L (RKO and HCT116) or 20 μmol/L (ARPE-19-MYC; DOX+) DAU alone or together with 50 nmol/L BAY-1895344 for 5 days. Representative images of spheroids are shown (left). Quantified spheroid sizes (right). n = 3. B, Nude mice bearing HCT116 xenograft tumors were treated with vehicle, DAU, and/or BAY-1895344 once daily, 5 days on/2 days off, intraperitoneally. The tumor volume was monitored and recorded. C and D, Tumors were excised at day 21 (C) and weighed (D). E, ARPE-19 cells expressing WT or the indicated mutant MYC were treated with DAU/BAY-1895344 for 24 hours and then subjected to apoptosis analysis. n = 4; *, P < 0.05; #, P < 0.0001. F, Apoptosis analysis of ARPE-19-MYCT58A treated with DAU/BAY-1895344 for 24 or 48 hours. n = 4; *, P < 0.05; #, P < 0.0001. G, ARPE-19-MYC (DOX+) and ARPE-19-MYC (DOX−) cells expressing HRASG12V or P53 shRNA were treated with DAU/BAY-189534 for 24 hours, followed by apoptosis analysis. H and I, Knockdown efficiency of P53 and the expression of HA-HRASG12V. n = 4; #, P < 0.0001. Graphic data are means ± SEM. For A and D, *, P < 0.05; **, P < 0.01; #, P < 0.0001; significantly different from vehicle-treated group. DOX, doxycycline; ctrl, control; hr, hours.

DAU and BAY-1895344 combination suppresses tumor growth 3D assays and in vivo.A, HCT116, RKO, and ARPE-19-MYC (DOX+) cells grown on Matrigel were treated with 5 μmol/L (RKO and HCT116) or 20 μmol/L (ARPE-19-MYC; DOX+) DAU alone or together with 50 nmol/L BAY-1895344 for 5 days. Representative images of spheroids are shown (left). Quantified spheroid sizes (right). n = 3. B, Nude mice bearing HCT116 xenograft tumors were treated with vehicle, DAU, and/or BAY-1895344 once daily, 5 days on/2 days off, intraperitoneally. The tumor volume was monitored and recorded. C and D, Tumors were excised at day 21 (C) and weighed (D). E, ARPE-19 cells expressing WT or the indicated mutant MYC were treated with DAU/BAY-1895344 for 24 hours and then subjected to apoptosis analysis. n = 4; *, P < 0.05; #, P < 0.0001. F, Apoptosis analysis of ARPE-19-MYCT58A treated with DAU/BAY-1895344 for 24 or 48 hours. n = 4; *, P < 0.05; #, P < 0.0001. G, ARPE-19-MYC (DOX+) and ARPE-19-MYC (DOX−) cells expressing HRASG12V or P53 shRNA were treated with DAU/BAY-189534 for 24 hours, followed by apoptosis analysis. H and I, Knockdown efficiency of P53 and the expression of HA-HRASG12V. n = 4; #, P < 0.0001. Graphic data are means ± SEM. For A and D, *, P < 0.05; **, P < 0.01; #, P < 0.0001; significantly different from vehicle-treated group. DOX, doxycycline; ctrl, control; hr, hours.