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Single-cell RNA-seq analysis of prostate cancer cells [II]

ABSTRACT: Androgen receptor (AR) plays key roles both in development of normal prostate gland and at all stages of prostate cancer. Due to the diversity of cell response to androgen stimulation, it’s necessary to investigate AR regulatory network at single cell level to fully understand how AR regulates transcription in prostate cancer cells. Here, we performed normal growing and time-series 5α-dihydrotestosterone (DHT) stimulated single cell RNA-seq profiling and transcriptome analysis on prostate cancer cell lines. We sequenced 186 single cell transcriptomes in total and found gene expression heterogeneity among each cell. Finally, novel biological processes have been discovered combined with clinical data. This study provides a comprehensive profile to dissect hormone regulation in prostate cancer at single cell resolution.

ORGANISM(S): Homo sapiens

PROVIDER: GSE145844 | GEO | 2021/10/10

REPOSITORIES: GEO

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Project description:Androgen receptor (AR) orchestrates an intricate transcriptional regulatory network that governs prostate cancer initiation, development and progression. To understand this network in detail, we generated genome-wide maps of AR occupancy by ChIP-seq in LNCaP cells. We found NKX3-1, an androgen-dependent homeobox protein well-characterized for its role in prostate development and differentiation, being recruited to AR binding sites (ARBS) in response to androgen signaling. We identified 6,359 NKX3-1 binding sites, most of which overlapped with AR. In addition to its novel collaborative transcriptional role at well-known prostate cancer model genes, our binding and knockdown studies further suggested that NKX3-1 potentially regulates AR in a feed-forward manner. Integrative analysis of Oncomine molecular concepts showed that these androgen-regulated AR and NKX3-1 associated genes are significantly overexpressed in prostate carcinoma as well as advanced and recurrent prostate tumors. From our transcriptomic profiling and Gene Ontology analysis, we observed that AR and NKX3-1 co-regulate genes involved in ‘protein trafficking’ processes, which are mandatory events in the integration of oncogenic signaling pathways leading to prostate cancer development and progression. Interestingly, we found that AR and NKX3-1 co-regulate several members of the RAB GTPase family of secretory/trafficking proteins via the involvement of FoxA1 in a ternary complex and we believe that these AR/NKX3-1/FoxA1 co-regulated RAB genes could serve as expression signatures in prostate carcinogenesis. More specifically, through functional analyses, we showed that NKX3-1, together with AR and FoxA1, could promote prostate cancer cell survival through activation of RAB3B expression. Collectively, our study has provided important insights into the hierarchical transcriptional regulatory network established between AR and NKX3-1 and sought to elucidate the important genetic-molecular-phenotypic paradigm in androgen-dependent prostate cancer. Gene expression profiling of LNCaP in response to ETOH (vehicle) or DHT (5α-dihydrotestosterone) stimulation across different treatment time-points using microarray.

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Single-cell RNA-seq analysis of prostate cancer cells [II]

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