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Updated: May 7, 2026
Sequencing Small Non-coding RNA from Formalin-fixed Tissues and Serum-derived Exosomes from Castration-resistant Prostate Cancer Patients
Published on: November 19, 2019
Next-generation sequencing of advanced prostate cancer treated with androgen-deprivation therapy
Prabhakar Rajan1, Ian M Sudbery2, M Eugenia M Villasevil1
1Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow, Cancer Research UK Beatson Institute, Glasgow, UK; Cancer Research UK Beatson Institute, The Beatson Institute for Cancer Research, Glasgow, UK.
Background:
Androgen-deprivation therapy (ADT) is standard treatment for locally advanced or metastatic prostate cancer (PCa). Many patients develop castration resistance (castration-resistant PCa [CRPC]) after approximately 2-3 yr, with a poor prognosis. The molecular mechanisms underlying CRPC progression are unclear.
Objective:
To undertake quantitative tumour transcriptome profiling prior to and following ADT to identify functionally important androgen-regulated pathways or genes that may be reactivated in CRPC.
Design, Setting, And Participants:
RNA sequencing (RNA-seq) was performed on tumour-rich, targeted prostatic biopsies from seven patients with locally advanced or metastatic PCa before and approximately 22 wk after ADT initiation. Differentially regulated genes were identified in treatment pairs and further investigated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) on cell lines and immunohistochemistry on a separate CRPC patient cohort. Functional assays were used to determine the effect of pathway modulation on cell phenotypes.
Outcome Measurements And Statistical Analysis:
We searched for gene expression changes affecting key cell signalling pathways that may be targeted as proof of principle in a CRPC in vitro cell line model.
Results And Limitations:
We identified ADT-regulated signalling pathways, including the Wnt/β-catenin signalling pathway, and observed overexpression of β-catenin in a subset of CRPC by immunohistochemistry. We validated 6 of 12 (50%) pathway members by qRT-PCR on LNCaP/LNCaP-AI cell RNAs, of which 4 (67%) demonstrated expression changes consistent with RNA-seq data. We show that the tankyrase inhibitor XAV939 (which promotes β-catenin degradation) reduced androgen-independent LNCaP-AI cell line growth compared with androgen-responsive LNCaP cells via an accumulation of cell proportions in the G0/G1 phase and reduction in the S and G2/M phases. Our biopsy protocol did not account for tumour heterogeneity, and pathway inhibition was limited to pharmacologic approaches.
Conclusions:
RNA-seq of paired PCa samples revealed ADT-regulated signalling pathways. Proof-of-principle inhibition of the Wnt/β-catenin signalling pathway specifically delays androgen-independent PCa cell cycle progression and proliferation and warrants further investigation as a potential target for therapy for CRPC.
Insights
Androgen-deprivation therapy (ADT) resistance in prostate cancer (PCa) involves reactivated signaling pathways. Targeting the Wnt/β-catenin pathway shows promise for treating castration-resistant PCa (CRPC).
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Androgen-deprivation therapy (ADT) is a standard treatment for advanced prostate cancer (PCa).
- Many patients develop castration-resistant prostate cancer (CRPC) with a poor prognosis.
- The molecular mechanisms driving CRPC progression remain unclear.
Purpose of the Study:
- To identify androgen-regulated pathways and genes reactivated in CRPC using transcriptome profiling.
- To investigate potential therapeutic targets for CRPC based on these findings.
Main Methods:
- RNA sequencing (RNA-seq) of paired PCa biopsies before and after ADT.
- Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and immunohistochemistry for validation.
- Functional assays using cell line models to assess pathway inhibition.
Main Results:
- ADT-regulated signaling pathways, including Wnt/β-catenin, were identified.
- Overexpression of β-catenin was observed in a subset of CRPC samples.
- Inhibition of Wnt/β-catenin signaling using XAV939 reduced androgen-independent PCa cell growth.
Conclusions:
- Transcriptome profiling revealed ADT-regulated pathways in PCa.
- The Wnt/β-catenin pathway is a potential therapeutic target for CRPC.
- Targeting this pathway demonstrates proof-of-principle for delaying CRPC progression.
