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. 2024 Apr 8;84(9):1426–1442. doi: 10.1158/0008-5472.CAN-23-3334

Figure 3.

Figure 3. Drug screen identifies palbociclib as a DSRCT therapeutic. A, Heatmap of 22 EWSR-WT1 upregulated genes with available inhibitors. B, Relative viability of JN-DSRCT-1 and LP9 cells treated with 1 or 10 μmol/L of various inhibitors. C, Relative viability of four DSRCT cell lines or LP9 mesothelial cells treated for 72 hours with palbociclib, KN93, UNC2250, vandetinib, or sunitinib (n = 3). D, Western blot analysis of MERTK, CAMK2A, CALM1, and CCND1 protein expression in shWT1 DSRCT cell lines treated with (+) or without (−) dox. Average protein expression was quantified relative to ACTIN, normalized to the -dox condition, and is shown underneath each blot (n = 2). E, Colony formation of JN-DSRT-1 and BER-DSRCT cells treated with palbociclib or abemaciclib for 12 days (n = 2). F, Western blot analysis of DSRCT cells treated with palbociclib for 2 days showing decreased phosphorylation or RB (representative image of n = 2). Phosphorylated RB fraction was determined by quantifying pRB S708 relative to total RB and normalizing to the untreated condition. Average protein expression of CDK4 and CDK6 was quantified relative to ACTIN and normalized to the untreated condition. Quantifications are shown underneath each blot. G, Cell-cycle analysis of DSRCT cells treated with palbociclib for 2 days (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Drug screen identifies palbociclib as a DSRCT therapeutic. A, Heatmap of 22 EWSR-WT1 upregulated genes with available inhibitors. B, Relative viability of JN-DSRCT-1 and LP9 cells treated with 1 or 10 μmol/L of various inhibitors. C, Relative viability of four DSRCT cell lines or LP9 mesothelial cells treated for 72 hours with palbociclib, KN93, UNC2250, vandetinib, or sunitinib (n = 3). D, Western blot analysis of MERTK, CAMK2A, CALM1, and CCND1 protein expression in shWT1 DSRCT cell lines treated with (+) or without (−) dox. Average protein expression was quantified relative to ACTIN, normalized to the -dox condition, and is shown underneath each blot (n = 2). E, Colony formation of JN-DSRT-1 and BER-DSRCT cells treated with palbociclib or abemaciclib for 12 days (n = 2). F, Western blot analysis of DSRCT cells treated with palbociclib for 2 days showing decreased phosphorylation or RB (representative image of n = 2). Phosphorylated RB fraction was determined by quantifying pRB S708 relative to total RB and normalizing to the untreated condition. Average protein expression of CDK4 and CDK6 was quantified relative to ACTIN and normalized to the untreated condition. Quantifications are shown underneath each blot. G, Cell-cycle analysis of DSRCT cells treated with palbociclib for 2 days (n = 3). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.