Identification of miR-30b-3p and miR-30d-5p as direct regulators of androgen receptor signaling in prostate cancer by complementary functional microRNA library screening

Abstract

The Androgen Receptor (AR) plays a key role in prostate biology and in the progression of prostate cancer (PCa) to castration resistance. The role of microRNAs (miRNAs) in aberrant AR signaling have not been fully characterized. Here we screened a library of 810 miRNA mimics to identify miRNAs that alter AR activity in complementary functional assays including protein lysate microarray (LMA) quantification of AR and PSA protein levels, AR transcriptional reporter activity, and AR-positive PCa cell viability. Candidate AR-regulating miRNAs were verified through AR transcriptional reporter and cell viability assays. MiRNA binding sites were found within the AR 3′-untranslated region (UTR) and within the AR and AR-V7 coding regions. MiRNA activity was characterized by western blotting, 3′-UTR reporter assay, and AR-GFP and AR-V7-GFP reporter assays. Results uncovered miR-30 family members as direct AR inhibitors. Inhibition of endogenous miR-30b-3p and miR-30d-5p enhanced AR expression and androgen-independent cell growth. Droplet digital RT-PCR quantification of miR-30c-5p and miR-30d-5p revealed significantly reduced levels in metastatic castration resistant PCa (CRPC), when compared to healthy prostate tissues. MiR-30d-5p levels were inversely correlated with AR activity, as measured by PSA mRNA, in metastatic CRPC. Collectively, these studies provide a comprehensive evaluation of AR-regulating miRNAs in PCa.

LDHA is a direct target of miR‐30d‐5p and contributes to aggressive progression of gallbladder carcinoma

“…31 In our study, high LDHA expression including metabolism in cancers. 32,33 There are many miRNAs targeting LDHA have been reported, such as miR-30a-5p, 34 40 Moreover, our data showed that miR-30d-5p over-expression inhibit glucose metabolism, cell proliferation, and tumorigenesis in GBC cell lines. This phenomenon could be phenolcopied by LDHA knockdown.…”

“…However, whether miRNA contributes to the dysregulated of LDHA in GBC has not been reported yet. miR-30d-5p could regulate androgen receptor signaling and exert a growth-suppressive role in prostate cancer 40. Meanwhile, we found downregulation of miR-30d-5p was frequently observed and negatively correlated with the LDHA expression in GBC tissues.…”

Expression Signature and Role of miR-30d-5p in Non-Small Cell Lung Cancer: a Comprehensive Study Based on in Silico Analysis of Public Databases and in Vitro Experiments

“…Previous studies have reported the involvement of miR-30d-5p in the carcinogenesis of several cancers, such as prostate cancer, colon cancer and conjunctival malignant melanoma [22][23][24]. Notably, miR-30d-5p expression is down-regulated in NSCLC tissues, and it exerts significant suppressive effects on the tumor growth, cell cycle progression, and motility of NSCLC cells [25].…”

Systematic Discovery of the Functional Impact of Somatic Genome Alterations in Individual Tumors through Tumor-specific Causal Inference

“…These SGA-FIs include the majority (87%) of the previously published drivers (1,2), as well as many novel candidate drivers. In addition to protein-coding genes, TCI also identified SGA events affecting microRNAs and intergenic non-protein-coding RNAs (e.g., M1R31HG(36,37), MIR30B(38), and PVT1(39’)) as SGA-FIs, (Supplementary Table S2).…”