The Mechanisms Of SP1 Mediated AR Signaling Pathway In The Progression Of Prostate Cancer

ObjectiveTo explore the molecular mechanism of SP1 in the AR transcriptional regulation pathway and the impact of SP1 on castration-resistant prostate cancer(CRPC).To explore the value of targeting SP1 in the treatment of CRPC.MethodsUsing ChIP-seq on SP1,AR,and RNA-seq data,we used the BETA(binding and expression targeting analysis)method to study whether SP1 participated in the AR transcription regulation network,and used q PCR Western blot,ChIP,and luciferase reporter gene experiments to verify the accuracy of the results.With the help of correlation analysis,cell and tissue immunofluorescence,protein interaction,and PLA assays,we combined these experiments to verify the interaction between SP1 and AR.To study the expression status of SP1 in CRPC in a low androgen environment,bioinformatics,CRISP-Cas9 technology,super enhancer interference experiments,and other methods were performed to analyze the mechanism of high expression of SP1 in CRPC.Through RNA-seq and pathway enrichment analysis on CRPC cell lines with SP1 silencing,the biological pathways and molecular mechanisms involved in SP1 in CRPC were studied.MTT,clone formation,Transwell,xenograft tumor model and other experiments were performed to study the effects of SP1 on the proliferation and invasion of CRPC cells.Finally,we used an antisense oligonucleotide(ASO)design process to complete the screening,synthesis and verification of antisense oligonucleotides specifically targeting SP1,and used in vitro and in vitro experiments to verify the therapeutic value of this ASO for CRPC.Results1.SP1 participates in AR-mediated transcriptional regulation functions(including transcriptional activation and transcriptional inhibition)in prostate cancer cells,and cooperates with AR to perform transcriptional regulation on KLK3 and other classic AR-target genes.During stimulation of androgen(DHT),SP1 and AR have protein-protein interactions.2.In CRPC,SP1 is highly expressed by the activation of a specific superenhancer,owing to epigenetic modifications but not genetic changes to the gene.3.SP1 participates in the regulation of CRPC cells by regulating the signaling pathways(such as PI3K/AKT and mTOR pathways).4.SP1 plays a key role in the growth and invasion of CRPC cells.SP1 can promote the proliferation and invasion of CRPC,suggesting that SP1 is an important therapeutic target for CRPC.5.Targeting SP1 has a clear therapeutic value for CRPC,and antisense oligonucleotides targeting SP1 are expected to become a new treatment for CRPC in the future.ConclusionsIn prostate cancer cells,when AR is activated by DHT stimulation,SP1 can interact with AR to co-regulate downstream target genes that are associated with AR.Upon entering the CRPC stage,SP1 will be highly expressed under the regulation of super enhancers.Additionally,due to the lack of DHT in the environment,SP1 will function independently.Combined with the integrated analysis of RNA-seq,we found that SP1 tends to activate PI3K/AKT and mTOR signaling pathways in CRPC,indicating that SP1 plays an important function in CRPC,and it also suggests that SP1 may become a drug target in CRPC with potential clinical value.