Journal: Cancer research

Article Title: Androgen receptor drives polyamine synthesis creating a vulnerability for prostate cancer

doi: 10.1158/0008-5472.CAN-25-1532

Figure Lengend Snippet: A. MYC ChIP-seq in VCaP cells treated with vehicle control or supraphysiological androgen (SPA; DHT 10nM) for 4 hours. Data reanalyzed from Guo et al, 2021( 40 ). B. Change in indicated transcript expression by RNAseq in VCaP cells with control knock-down or MYC knock-down by siRNA. Data reanalyzed from Guo et al, 2021( 40 ). C . Expression of indicated proteins in LNCaP cells expressing empty vector (EV) or MYC (MYC) treated with vehicle control (V; EtOH 0.1%) or SPA (S; R1881 10nM) for 96 hours. Vinculin is used as a loading control. Representative blot of n=2 experiments. D. Change in indicated transcript expression by RNAseq in cells treated as per C. E. A comparison of change of individual RNA transcript abundance by SPA in LNCaP-EV cells versus LNCaP-MYC cells. The line of best fit (dotted red) has a shallower slope than the line of unity (solid blue), indicating that many transcripts have reduced change by SPA in LNCaP-MYC cells. F. Schematic highlighting that the inhibition of MYC by SPA can function as an amplifying circuit to further increase expression of ODC1 , AMD1 , and KLK3 by SPA. P values by unpaired 2-tailed t test in B and D.

Article Snippet: RT-PCR were performed in triplicate using 500ug cDNA, 10ul TaqMan Gene Expression Master Mix (ThermoFisher), and 1ul 20x TaqMan Gene Expression Assay probe mix for MYC (Hs00153408_m1), AMD1 (Hs00750876_s1), ODC1 (Hs00159739_m), KLK3 (Hs02576345_m1) and ACTB (Hs01060665_g1) (ThermoFisher) on an ABI7500 RT-PCR System (ThermoFisher).

Techniques: Expressing, ChIP-sequencing, Control, Knockdown, Plasmid Preparation, Comparison, Inhibition

Journal: Cancer research

Article Title: Androgen receptor drives polyamine synthesis creating a vulnerability for prostate cancer

doi: 10.1158/0008-5472.CAN-25-1532

Figure Lengend Snippet: A. Protein expression of ODC in LNCaP cells expressing dCas9-KRAB and sgRNA-C or sgRNA-D targeting site 2 upstream of ODC1 with and without concurrent constitutive ODC1 cDNA expression treated with vehicle control (V; EtOH 0.1%) or supraphysiological androgen (S; R1881 10nM) for 6 days. Vinculin is used as a loading control. B. Putrescine abundance in media of cells treated as per A. C. Viable cell number relative to vehicle treatment of cells as per A. D. Relative viable cell number of cells treated with combinations of vehicle control (VEH; EtOH 0.1%), supraphysiological androgen (SPA; R1881 10nM), difluoromethylornithine (DFMO 5mM), and putrescine (PUT 100uM) for 7 days. E. MycCaP-CR tumor size over time following treatment with control (empty pellet), SPA (testosterone pellet), DFMO (2% in drinking water), or SPA and DFMO. *p<0.05, **p<0.01, ***p<0.001 by unpaired t test of tumor volumes on day 32. F. ODC activity of MycCaP-CR tumors treated for 14 days as per E. P value by unpaired 2-tailed t test in B-D and F. N = 3 independent experiments performed in A-D.

Article Snippet: RT-PCR were performed in triplicate using 500ug cDNA, 10ul TaqMan Gene Expression Master Mix (ThermoFisher), and 1ul 20x TaqMan Gene Expression Assay probe mix for MYC (Hs00153408_m1), AMD1 (Hs00750876_s1), ODC1 (Hs00159739_m), KLK3 (Hs02576345_m1) and ACTB (Hs01060665_g1) (ThermoFisher) on an ABI7500 RT-PCR System (ThermoFisher).

Techniques: Inhibition, Expressing, Control, Activity Assay

Journal: Neoplasia (New York, N.Y.)

Article Title: High-dose DFMO alters protein translation in neuroblastoma

doi: 10.1016/j.neo.2025.101215

Figure Lengend Snippet: ODC1 and MYCN co-occurring amplification. (A) Neuroblastoma cell lines with distinct genomic profiles are shown: NBLS cells have neither MYCN nor ODC1 amplification, IMR5 has MYCN amplification (and ALK amplification, not shown), and KCN has both MYCN and ODC1 amplification; data represents Log R ratio output from lllumina Bead-Chip SNP arrays. (B) Fluorescence in situ hybridization (FISH) results for a primary neuroblastoma sample using probes for MYCN (2p24.3, red), ODC1 (2p25.1, aqua) and 2p centromere (CEN2, green) showing 4 CEN2 signals and amplification of both MYCN and ODC1 . Images courtesy of CHOP Division of Genomic Diagnostics (DGD).

Article Snippet: Primers: MTTP assay #Hs04877005_cn, ODC1 assay #Hs02651018_cn, and MYCN assay #Hs00201049_cn (Applied Biosystems), chosen for similar amplicon size (80-100 bps) for matched amplification efficiency.

Techniques: Amplification, Fluorescence, In Situ Hybridization

Journal: Neoplasia (New York, N.Y.)

Article Title: High-dose DFMO alters protein translation in neuroblastoma

doi: 10.1016/j.neo.2025.101215

Figure Lengend Snippet: DFMO-mediated translation effects. (A) Phospho-4EBP1 is not altered by DFMO (5 uM) but is reduced by the mTOR1/2 inhibitor, MLN0128 (100 μ M). (B) More sensitive IEF immunoblot confirms incomplete hypusination in most cell lines at higher DFMO exposures. (C) Dose response to DFMO for IMR5 and SK-N-BE2C cells (cells with higher proportion of non-hypusinated eIF5A in (B)). Cell line genomic status: M , MYC amplification; N , MYCN amplification; O , ODC1 amplification.

Article Snippet: Primers: MTTP assay #Hs04877005_cn, ODC1 assay #Hs02651018_cn, and MYCN assay #Hs00201049_cn (Applied Biosystems), chosen for similar amplicon size (80-100 bps) for matched amplification efficiency.

Techniques: Western Blot, Amplification

Journal: Neoplasia (New York, N.Y.)

Article Title: High-dose DFMO alters protein translation in neuroblastoma

doi: 10.1016/j.neo.2025.101215

Figure Lengend Snippet: Puromycin incorporation by DFMO exposure in vitro. Puromycin incorporation was assessed across neuroblastoma cell lines of distinct MYCN and ODC1 gene status, as denoted. Densitometry was used to define relative change from vehicle control lanes; replicates are shown in line graph form with the immunoblots corresponding to the data from the circle-marked line. Cell line genomic status: M , MYC amplification; N , MYCN amplification; O , ODC1 amplification.

Article Snippet: Primers: MTTP assay #Hs04877005_cn, ODC1 assay #Hs02651018_cn, and MYCN assay #Hs00201049_cn (Applied Biosystems), chosen for similar amplicon size (80-100 bps) for matched amplification efficiency.

Techniques: In Vitro, Control, Western Blot, Amplification

Journal: Neoplasia (New York, N.Y.)

Article Title: High-dose DFMO alters protein translation in neuroblastoma

doi: 10.1016/j.neo.2025.101215

Figure Lengend Snippet: Impact of AMXT-1501 and DFMO on puromycin incorporation. Puromycin incorporation was assessed across neuroblastoma cell lines of distinct MYCN and ODC1 gene status, as denoted, treated with both DFMO and AMXT-1501. Densitometry was used to define relative change from vehicle control lanes; replicates are shown in line graphs with the immunoblots corresponding to the data from the cirlce-marked line. Cell line genomic status: M , MYC amplification; N , MYCN amplification; O , ODC1 amplification.

Article Snippet: Primers: MTTP assay #Hs04877005_cn, ODC1 assay #Hs02651018_cn, and MYCN assay #Hs00201049_cn (Applied Biosystems), chosen for similar amplicon size (80-100 bps) for matched amplification efficiency.

Techniques: Control, Western Blot, Amplification

Journal: Neoplasia (New York, N.Y.)

Article Title: High-dose DFMO alters protein translation in neuroblastoma

doi: 10.1016/j.neo.2025.101215

Figure Lengend Snippet: Colony Formation Assay across a range of DFMO exposures. Colony formation was assessed across neuroblastoma cell lines of distinct MYCN and ODC1 gene status, as denoted, treated with DFMO across a range of concentrations. Data represents replicate wells with representative well-images shown above. Cell line genomic status: M , MYC amplification; N , MYCN amplification; O , ODC1 amplification; *= p < 0.05, **= p < 0.01, ***= p < 0.001.

Article Snippet: Primers: MTTP assay #Hs04877005_cn, ODC1 assay #Hs02651018_cn, and MYCN assay #Hs00201049_cn (Applied Biosystems), chosen for similar amplicon size (80-100 bps) for matched amplification efficiency.

Techniques: Colony Assay, Amplification

Journal: Neoplasia (New York, N.Y.)

Article Title: High-dose DFMO alters protein translation in neuroblastoma

doi: 10.1016/j.neo.2025.101215

Figure Lengend Snippet: ODC protein in response to DFMO exposure. (A, B) Treatment with DFMO leads to increased ODC protein levels by immunoblot across cell lines. (C) Dose response of IMR5 and CHLA136 cells to DFMO: ODC is increased at baseline in the ODC1- amplified cell line without significant increase in response to DFMO; shown as immunoblot and by densitometry relative to control lanes. Cell line genomic status: M , MYC amplification; N , MYCN amplification; O , ODC1 amplification.

Article Snippet: Primers: MTTP assay #Hs04877005_cn, ODC1 assay #Hs02651018_cn, and MYCN assay #Hs00201049_cn (Applied Biosystems), chosen for similar amplicon size (80-100 bps) for matched amplification efficiency.

Techniques: Western Blot, Amplification, Control