Fig 1.
ODC1 expression in endometrial cancers.
A-C in LCM (laser capture micro-dissected) samples. A, Endometrioid (E, n = 139) and Serous (S, n = 37) types and normal epithelial tissues (N, n = 12); B, FIGO Stages I and II (n = 133), III (n = 24), and IV (n = 18); C, Grade 1 (G1, n = 42), Grade 2 (G2, n = 65), and Grade 3 (G3, n = 69). Signal: Affymetrix signal normalized to target value of 500. D, In TCGA samples of four molecular sub-types: Copy number low (MSS, n = 90), MSI high (n = 65), POLE ultra-mutant (n = 17) and Copy number high—serous like (n = 60). ***p < 0.001; **p < 0.01; *p< 0.05. Counts: Normalized RNA-seq counts.
Fig 2.
ODC1 mRNA expression overall survival (OS) and recurrence free interval (RFI).
Kaplan-Meier diagrams and Wald statistics showing increased ODC1 is significantly associated with shorter OS (A) in TCGA (RNA-seq) (n = 232), LCM cases (Affymetrix) (n = 188) and in Spectrum (qRT-PCR) (n = 60) samples (B) ODC1 mRNA is significantly associated with recurrence for TCGA and qRT-PCR cohorts but is not significant in the Affymetrix LCM dataset. “High” and “Low” indicate above and below the median expression level.
Fig 3.
Bar graph of ODC1 mRNA expression overall survival (OS) and recurrence free interval (RFI).
A, Bar graph indicates relative expressions of OS in TCGA (RNA-seq) (p = 0.00352), LCM cases (Affymetrix) (p = 1.3e-05) and in Spectrum (qRT-PCR) (p = 0.00846) samples. B, Relative expressions of RFI in TCGA (RNA-seq) (p = 0.0248), LCM cases (Affymetrix) (p = 0.211) and in Spectrum (qRT-PCR) (p = 0.00295) samples. Rec.: Recurrence; NED: No Evidence of Disease.
Fig 4.
Colony formation, cell viability and polyamine levels in DFMO-treated endometrial cancer cells.
A, Colony counts in a panel of endometrial cancer and immortal normal endometrial epithelial cells including ACI-98 (undifferentiated), MSU-15 (clear cell), ACI-61 (endometrioid) ACI-70 (MMT), HEC-1-A (endometrioid), EM E6/E7 TERT1 (normal immortalized endometrial epithelial), ECC-1 (endometrioid) and ACI-45 (carcinosarcoma, MMT). Cells are ordered most highly sensitive on left to least sensitive on the right. B, Representative colony plates for most (ACI-98) and least (ACI-45) sensitive cells. C and D, ACI-98 cells highly sensitive to wide range of doses of DFMO while ACI-45 cells are non-sensitive even at very high doses (5 mM) as measured by MTS assay. The percent values (%) shown are DFMO to Control. Intracellular polyamine levels (putrescine, spermidine, spermine) were analyzed in ACI-98 (E) and ACI-45 (F) cells either untreated (control) or DFMO-treated (0.25 mM) using RP-HPLC. Intracellular polyamines were quantified and expressed as nmol/mg protein. Standard errors are indicated (+/- S.E.). Put, putrescine; Spd, spermidine; Spm, spermine.
Fig 5.
Oral DFMO significantly reduces tumor burden in tumor-bearing mice.
Female athymic nude mice bearing xenografted endometrial tumors (ACI-98) received 2% (w/v) DFMO in drinking water or control (water only). Both control and DFMO-treated mice consumed similar quantities of water per day. Quantification of tumor volume (A) and tumor weight (B) of DFMO-treated and untreated tumors. Ten mice per group (n = 10). C, Representative images of DFMO-treated and untreated tumors.
Fig 6.
Effect of oral DFMO on polyamine levels in tumor-bearing mice.
Intracellular polyamine levels (putrescine, spermidine, spermine) were analyzed in tumor tissue (A) and blood plasma (B) of DFMO-treated and untreated mice using reverse-phase HPLC. Intracellular polyamines were quantified and expressed as nmol/mg protein (A) or nmol/ml plasma (B). Standard errors are indicated (+/- S.E.). Put, putrescine; Spd, spermidine; Spm, spermine.