title: PCOTH, a novel gene overexpressed in prostate cancers, promotes prostate cancer cell growth through phosphorylation of oncoprotein TAF-Ibeta/SET.
abstract: Through genome-wide cDNA microarray analysis coupled with microdissection of prostate cancer cells, we identified a novel gene, prostate collagen triple helix (PCOTH), showing overexpression in prostate cancer cells and its precursor cells, prostatic intraepithelial neoplasia (PIN). Immunohistochemical analysis using polyclonal anti-PCOTH antibody confirmed elevated expression of PCOTH, a 100-amino-acid protein containing collagen triple-helix repeats, in prostate cancer cells and PINs. Knocking down PCOTH expression by small interfering RNA (siRNA) resulted in drastic attenuation of prostate cancer cell growth, and concordantly, LNCaP derivative cells that were designed to constitutively express exogenous PCOTH showed higher growth rate than LNCaP cells transfected with mock vector, suggesting the growth-promoting effect of PCOTH on prostate cancer cell. To investigate the biological mechanisms of this growth-promoting effect, we applied two-dimensional differential gel electrophoresis (2D-DIGE) to analyze the phospho-protein fractions in LNCaP cells transfected with PCOTH. We found that the phosphorylation level of oncoprotein TAF-Ibeta/SET was significantly elevated in LNCaP cells transfected with PCOTH than control LNCaP cells, and these findings were confirmed by Western blotting and in-gel kinase assay. Furthermore, knockdown of endogenous TAF-Ibeta expression by siRNA also attenuated viability of prostate cancer cells as well. These findings suggest that PCOTH is involved in growth and survival of prostate cancer cells thorough, in parts, the TAF-Ibeta pathway, and that this molecule should be a promising target for development of new therapeutic strategies for prostate cancers.
authors.affiliation: Laboratory of Molecular Medicine, Human Genome Center, Institute of Medical Science, University of Tokyo, Tokyo, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Anazawa
authors.forename: Hidewaki
authors.initials: H
authors.lastname: Nakagawa
authors.forename: Mutsuo
authors.initials: M
authors.lastname: Furihara
authors.forename: Shingo
authors.initials: S
authors.lastname: Ashida
authors.forename: Kenji
authors.initials: K
authors.lastname: Tamura
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yoshioka
authors.forename: Taro
authors.initials: T
authors.lastname: Shuin
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Fujioka
authors.forename: Toyomasa
authors.initials: T
authors.lastname: Katagiri
authors.forename: Yusuke
authors.initials: Y
authors.lastname: Nakamura
mesh_terms: D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D002868:Chromosomal Proteins, Non-Histone / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D003094:Collagen / Q000096:biosynthesis / Q000235:genetics*; D004268:DNA-Binding Proteins; D015180:Electrophoresis, Gel, Two-Dimensional; D015870:Gene Expression; D056488:Histone Chaperones; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D034741:RNA, Small Interfering / Q000235:genetics; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D004268:DNA-Binding Proteins; D056488:Histone Chaperones; D034741:RNA, Small Interfering; C513473:SET protein, human; D014157:Transcription Factors; D003094:Collagen