RRC ID 45571
Author Guo W, Keener AL, Jing Y, Cai L, Ai J, Zhang J, Fisher AL, Fu G, Wang Z.
Title FOXA1 modulates EAF2 regulation of AR transcriptional activity, cell proliferation, and migration in prostate cancer cells.
Journal Prostate
Abstract BACKGROUND:ELL-associated factor 2 (EAF2) is an androgen-regulated tumor suppressor in the prostate. However, the mechanisms underlying tumor suppressive function of EAF2 are still largely unknown. Identification of factors capable of modulating EAF2 function will help elucidate the mechanisms underlying EAF2 tumor suppressive function.
METHODS:Using eaf-1(the ortholog of EAF2) mutant C. elegans model, RNAi screen was used to identify factors on the basis of their knockdown to synergistically enhance the reduced fertility phenotype of the eaf-1 mutant C. elegans. In human cells, the interaction of EAF2 with FOXA1 and the effect of EAF2 on the FOXA1 protein levels were determined by co-immunoprecipitation and protein stability assay. The effect of EAF2 and/or FOXA1 knockdown on the expression of AR-target genes was determined by real-time RT-PCR and luciferase reporter assays. The effect of EAF2 and/or FOXA1 knockdown on LNCaP human prostate cancer cell proliferation and migration was tested using BrdU assay and transwell migration assay.
RESULTS:RNAi screen identified pha-4, the C. elegans ortholog of mammalian FOXA1, on the basis of its knockdown to synergistically enhance the reduced fertility phenotype of the eaf-1 mutant C. elegans causing sterility. EAF2 co-immunoprecipitated with FOXA1. EAF2 knockdown enhanced endogenous FOXA1 protein level, whereas transfected GFP-EAF2 down-regulated the FOXA1 protein. Also, EAF2 knockdown enhanced the expression of AR-target genes, cell proliferation, and migration in LNCaP cells. However, FOXA1 knockdown inhibited the effect of EAF2 knockdown on AR-target gene expression, cell proliferation, and migration in LNCaP cells, suggesting that FOXA1 can modulate EAF2 regulation of AR transcriptional activation, cell proliferation, and migration.
CONCLUSIONS:These findings suggest that regulation of the AR signaling pathway, cell proliferation, and migration through FOXA1 represents an important mechanism of EAF2 suppression of prostate carcinogenesis.
Volume 75(9)
Pages 976-87
Published 2015-6-15
DOI 10.1002/pros.22982
PMID 25808853
PMC PMC4424106
MeSH Animals Blotting, Western Caenorhabditis elegans Cell Line, Tumor Cell Proliferation / physiology Down-Regulation Hepatocyte Nuclear Factor 3-alpha / genetics Hepatocyte Nuclear Factor 3-alpha / metabolism* Humans Immunoprecipitation Male Prostatic Neoplasms / genetics Prostatic Neoplasms / metabolism* Prostatic Neoplasms / pathology RNA Interference / physiology Receptors, Androgen / genetics Receptors, Androgen / metabolism* Transcription Factors / genetics Transcription Factors / metabolism* Transcriptional Activation Transfection / methods
IF 3.279
Times Cited 15
WOS Category UROLOGY & NEPHROLOGY ENDOCRINOLOGY & METABOLISM
Resource
C.elegans tm3976