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. 2024 Oct 29;221(11):e20231832. doi: 10.1084/jem.20231832

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© 2024 Hsu et al.

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Figure S4. — IMPA1 and IMPDH2 are upregulated in prostate cancer patients correlated with poor survival outcome. (A) The frequency of alteration of IMPA1 gene in human prostate cancers. The representative data obtained from cBioPortal (https://www.cbioportal.org) representing a combined study of 4,104 samples, querying 3,886 prostate cancer patients in 16 studies are shown by a bar graph. (B) The bar graph represents the relative expression of IMPA1 in normal tissues (152) and prostate cancers (492) from the GEPIA database. P < 0.01. (C) The overall survival of prostate cancer patients with IMPA1 altered group (121 cases) and IMPA1 unaltered group (1,157 cases). The representative data also obtained from cBioPortal (https://www.cbioportal.org) representing a combined study of 4,104 samples, querying 3,886 prostate patients in 16 studies as shown by overall survival Kaplan–Meier plot. The median months overall survival of the IMPA1 altered group and IMPA1 unaltered group are 97 and 141 mo, respectively. Long–rank test P value = 3.69e−8. (D) The overall survival Kaplan–Meier plot of prostate cancer patients with IMPA1 high and low expression from the database of PrognoScan. Long–rank test P value = 0.045595. (E) Prostate tissue from AR⁺ metastatic prostate cancer and AR^low/− metastatic prostate cancer patients in our in-house prostate cancer samples were determined by IHC staining with indicated antibodies. Scale bar, 100 μm. (F and G) Box plot represents IMPA1 (F) and IMPDH2 (G) expression in AR⁺ metastatic prostate cancer (20 cases) and AR⁻ metastatic prostate cancer (20 cases). ***, P < 0.001 by Mann–Whitney U test. (H) Prostate tissue from AR⁺ CRPC and AR^low/− CRPC in our in-house prostate cancer samples were determined by IHC staining with indicated antibodies. Scale bar, 200 μm. (I) Box plot represents IMPA1 expression levels by H-score in AR⁺ CRPC (n = 5) and AR^low/− CRPC (n = 15) prostate cancer patients. ***, P < 0.001 (AR⁺ CRPC versus AR^low/- CRPC) by Mann–Whitney U test. (J) Inositol levels were determined by LC-MS/MS in the prostate tissues of the patients without or with the development to CRPC. ***, P < 0.001 (patients without CRPC versus patient with CRPC) by Mann–Whitney U test. (K) Soft agar assay shown in 22RV1 cells stably expressing shLuc or shIMPDH2 (#1 and #2). (L) Quantification of the number of colonies from 22RV1 by ImageJ shown in K. The data were shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01 by two-tailed unpaired t test. (M) Soft agar assay shown in 22RV1 cells stably expressing shLuc or shIMPDH2 upon the treatment of 40 μM of guanosine (Gua), 40 μM of inositol (Ins) or 40 μM of adenosine (Ade) for 20 days. (N) Quantification of the number of colonies from 22RV1 by ImageJ shown in M. The data were shown as the mean ± SEM of three independent experiments for each group. *, P < 0.05; **, P < 0.01 by two-tailed unpaired t test. (O) Male nude mice subcutaneously injected with 22RV1 cells stably expressing shLuc or shIMPDH2 were intraperitoneally injected with 30 mg/kg of inositol (Ins) or 30 mg/kg of guanosine (Gua) every 2 days until 39 days, and tumor volume was measured with indicated days. Four mice were included in each experimental group. (P) IMPDH2 protein expression from adjacent normal and prostate cancer patients with high Gleason scores, and advanced stage in our in-house prostate cancer samples was determined by IHC staining. Scale bar, 20 μm. (Q) Box plot represents IMPDH2 expression in adjacent normal (80 cases) and prostate cancer patients with high Gleason score and advanced stage (80 cases). ***, P < 0.001 by Mann–Whitney U test. (R) Disease-specific survival outcome with IMPDH2 low or high expression was shown by Kaplan–Meier plots in prostate cancer patients with high Gleason score and advanced stage (80 cases). *, P = 0.0171 by long–rank test.