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Effect of depletion of PRCAT71 and KHSRP on gene expression in prostate cancer cells

ABSTRACT: effect of depletion of PRCAT71 and KHSRP on gene expression in prostate cancer cells

PROVIDER: PRJNA1002000 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25510288_1.fastq.gz	Fastqsanger.gz
	SRR25510288_2.fastq.gz	Fastqsanger.gz
	SRR25510289_1.fastq.gz	Fastqsanger.gz
	SRR25510289_2.fastq.gz	Fastqsanger.gz
	SRR25510290_1.fastq.gz	Fastqsanger.gz

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effect of depletion of PRCAT71 and KHSRP on gene expression in prostate cancer cells

Project description:Mounting evidence indicates that long non-coding RNAs (lncRNAs) play vital roles in tumorigenesis and progression of cancers. However, the functions and regulatory mechanisms of lncRNAs in prostate cancer (PCa) are still largely unknown. In this study, we discovered a PCa-specific lncRNA, PRCAT71, significantly highly expressed in metastatic and primary PCa compared to benign prostate tissues. Silencing PRCAT71 inhibited cancerous properties of PCa cells, as well as androgen receptor (AR) signaling. Mechanismly, PRCAT71 acts as a scaffold to recruit KH-type splicing regulatory protein (KHSRP) to AR mRNA and stabilize AR mRNA. Furthermore, PRCAT71 is transcriptionally regulated by AR, thus forming a positive regulatory loop between AR and PRCAT71 in PCa. Our findings demonstrate that the PRCAT71-KHSRP-AR axis is a promising therapeutic target to treat PCa.

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Project description:We established LNCaP cells in wihch KHSRP was knocked down along with LNCaP cells with silenced KHSRP and re-introduced HA-tagged KHSRP-WT/K205R.RNA-seq was carried out in the stable LNCaP cell lines to determine the underlying mechanism of KHSRP acetylation in PCa tumorigenesis. We then performed gene expression profiling analysis using data obtained from RNA-seq of 4 different cells.

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Effect of KHSRP acetylation on gene expression in LNCaP cells

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Project description:Although the vital role of the androgen receptor (AR) has been well demonstrated in primary prostate cancers, its role in the androgen-insensitive prostate cancers still remains unclear. Here, we used a small hairpin RNA approach to directly assess AR activity in prostate cancer cells. Reduction of AR expression in the two androgen-sensitive prostate cancer cell lines, LNCaP and LAPC4, significantly decreased AR-mediated transcription and cell growth. Intriguingly, in two androgen-insensitive prostate cell lines, LNCaP-C42B4 and CWR22Rv1, knockdown of AR expression showed a more pronounced effect on AR-induced transcription and cell growth than androgen depletion. Using cDNA microarrays, we also compared the transcriptional profiles induced by either androgen depletion or AR knockdown. Although a significant number of transcripts appear to be regulated by both androgen depletion and AR knockdown, we observed a subset of transcripts affected only by androgen depletion but not by AR knockdown, and vice versa. Finally, we demonstrated a direct role for AR in promoting tumor formation and growth in a xenograft model. Taken together, our results elucidate an important role for the AR in androgen-insensitive prostate cancer cells, and suggest that AR can be used as a therapeutic target for androgen-insensitive prostate cancers.

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RNA immunoprecipitation (RIP) of KHSRP from pancreatic cancer cell extracts cultured on fibronectin

Project description:KHSRP-bound transcripts were identified by RNA immunoprecipitation and sequencing (RIP-Seq) and bioinformatics analysis. A pancreatic cancer cell line S2-013 was cultured on fibronectin. We performed RIP of KHSRP using anti-KHSRP antibody from the fibronectin-stimulated S2-013 extracts, and identified the associated RNAs using a next generation sequencer.

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Effect of androgen induction and AR depletion on gene expression of metastatic prostate cancer-specific long noncoding RNAs in prostate cancer cells.

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