AT‐rich interaction domain 5A regulates the transcription of interleukin‐6 gene in prostate cancer cells

• Published in: The Prostate 2022-01, Vol.82 (1), p.97-106
• Main Authors: Ikeuchi, Wataru, Wakita, Yuriko, Zhang, Guoxiang, Li, Chun, Itakura, Keiichi, Yamakawa, Takahiro
• Format: Article
• Language: English
• Subjects: ARID5A; Cloning; CRISPR; CRISPR‐SAM; Cytokines; DNA microarrays; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; DNA-directed RNA polymerase; Down-Regulation; Gene Expression Regulation; Gene Knockout Techniques - methods; Gene regulation; Histone Code; histone modification; Histones; Humans; IL‐6; Interleukin-6 - genetics; Interleukin-6 - metabolism; Male; Molecular modelling; mRNA stability; PC-3 Cells; Prostate cancer; Prostatic Neoplasms - genetics; Prostatic Neoplasms - metabolism; Protein Array Analysis - methods; RNA polymerase; RNA Stability; Statistical analysis; Transcription Factors; transcriptional regulation
• ISSN: 0270-4137; 1097-0045
• DOI: 10.1002/pros.24251

Background Interleukin‐6 (IL‐6) is a pleiotropic cytokine that confers androgen‐independence and aggressiveness in prostate cancer (PCa); however, the molecular mechanisms regulating IL‐6 expression remain unclear. The expression of ARID5A, an AT‐rich interaction domain (ARID) DNA‐binding motif‐containing transcription factor is positively correlated with IL‐6 expression in human PCa. We, therefore, hypothesized that ARID5A could regulate IL‐6 expression in PCa. Methods The relationship between ARID5A and IL‐6 in PCa patients was analyzed using statistical analyses of multiple clinical microarray data sets. To investigate whether ARID5A regulates IL‐6 expression, CRISPR‐driven ARID5A knockout clones were established in DU145 and PC‐3 cells. Results Analysis of three microarray data sets showed a positive correlation between ARID5A and IL‐6 expression. The expression of IL‐6 in ARID5A knockout clones was significantly reduced compared with control clones in both PCa cell lines. Knockout of ARID5A did not result in any loss of IL‐6 mRNA stability. Instead, we observed a significant decrease in the occupancy of both active RNA Polymerase II and the active histone mark, H3K4me3 at the IL‐6 transcriptional start site in ARID5A knockout PCa cells, suggesting a role for transcriptional regulation. Conclusions Our study demonstrated that loss of ARID5A downregulates the expression of IL‐6 at the transcriptional level.