ARRDC4 and UBXN1: Novel Target Genes Correlated with Prostate Cancer Gleason Score

SIMPLE SUMMARY: Prostate cancer (PCa) is the second most diagnosed malignancy in men. PCa is a heterogeneous disease, with the clinical presentation ranging from localized and indolent to a rapidly progressing lethal metastatic disease, therefore, we needed the predictor to diagnosis of aggressive PCa. The most important predictor of PCa outcomes has been demonstrated to be the histological Gleason score which was confirmed after prostate biopsy. In this study, we found that germline variants of ARRDC4 and UBXN1 could affect the prostate cancer Gleason score, which could be a potential marker to select aggressive PCa. ABSTRACT: To investigate potential markers of the prostate cancer (PCa) Gleason score (GS), genetic arrays in 841 PCa patients were conducted followed by functional validation in PCa cell lines. A total of 841 PCa patients who received radical prostatectomy (RP) from November 2003 to July 2019 were enrolled. HumanExome BeadChip 12v1-1 (Illumina, Inc.; San Diego, CA, USA) exomic arrays were performed on RP tissue samples. Unconditional logistic regression was used to calculate odds ratios to generate estimates of the relative risk of pathologic GS (≥8); SNPs with the highest association were selected and validated using PCa cell lines (PC3, LNCaP, 22Rv1 and DU145). Following transfection with target-gene siRNA, assays for cell viability, wound healing, and transwell invasion were performed. Mean age of enrolled subjects was 66.34 years and median PSA was 8.43 ng/mL. After RP, 122 patients (14.5%) had pathological Gleason scores ≥8. The results from genotyping with 242,186 SNPs by exomic array revealed that 4 SNPs (rs200944490, rs117555780, rs34625170, and rs61754877) were significantly associated with high pathological GS (≥8) within cut-off level to p < 10(−5). The most highly associated rs200944490 in ARRDC4 (p = 1.39 × 10(−6)) and rs117555780 in UBXN1 (p = 2.92 × 10(−5)) were selected for further validation. The knockdown of UBXN1 and ARRDC4 led to significantly reduced cell proliferation and suppressed migration and invasiveness in PCa cell lines. Epithelial mesenchymal transition (EMT) markers were significantly down-regulated in si-ARRDC4 and si-UBXN1-transfected cells. The expression levels of PI3K-phosphorylation and Akt phosphorylation and NF-κB were also suppressed following knockdown of UBXN1 and ARRDC4. The rs200944490 (ARRDC4) and rs117555780 (UBXN1) were identified as candidate markers predictive of PCa Gleason score which is strongly associated with cancer aggressiveness. Additional validation in future studies is warranted.