title: Production of Cyr61 protein is modulated by extracellular acidification and PI3K/Akt signaling in prostate carcinoma PC-3 cells.
abstract: High expression of Cyr61, an extracellular cysteine-rich heparin-binding protein, has been associated with a malignant cell phenotype and poor outcome in prostate cancers. Although Cyr61 was found by us to be overproduced in androgen-independent PC-3 cells treated with N-acetylcysteine (NAC), its significance is still unclear. We therefore aimed to determine how and why Cyr61 protein is overexpressed in NAC-treated cells. Here, we found that Cyr61 protein level markedly increased in cells treated with NAC at high cell seeding density. Silencing of Cyr61 by siRNA induced enhanced activity of caspase-3/7, upregulation of the proapototic Bok, BimL and BimS, cleavage of apoptosis hallmarkers such as Bax, PARP and caspase-3, and downregulation of antiapoptotic Bcl2, Bcl-xL and Mcl-1 proteins. NAC treatment caused a reduction of extracellular medium pH to acidic and an increase in Akt phosphorylation, after which the replacement with NAC-free medium returned them to control levels within 24h. Acid stimulation increased the levels of Cyr61 and p-Akt proteins, whereas it suppressed the induction of proapoptotic and antiapoptotic proteins. Overall, our data indicate that PC-3 cells overproduce Cyr61 protein via activation of the PI3K/Akt signaling as a part of the survival mechanisms under the conditions causing extracellular acidity and further cytotoxicity.
authors.affiliation: Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Soonchunhyang University, Cheonan 330-930, Republic of Korea.
authors.forename: Yoon-Jin
authors.initials: YJ
authors.lastname: Lee
authors.forename: David M
authors.initials: DM
authors.lastname: Lee
authors.forename: Sang-Han
authors.initials: SH
authors.lastname: Lee
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000143:Acids / Q000378:metabolism*; D017209:Apoptosis; D045744:Cell Line, Tumor; D055515:Cysteine-Rich Protein 61 / Q000096:biosynthesis* / Q000235:genetics; D020868:Gene Silencing; D006801:Humans; D008297:Male; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction; D015854:Up-Regulation
chemical_list: D000143:Acids; C501848:CCN1 protein, human; D055515:Cysteine-Rich Protein 61; D019869:Phosphatidylinositol 3-Kinases; D051057:Proto-Oncogene Proteins c-akt; D000111:Acetylcysteine
keywords: Akt; Apoptotic cell death; Cyr61; ECL; ECM; Extracellular acidity; HRP; N-acetylcysteine; NAC; PBS; PI3K; Prostate cancer; SDS; c-FLIP; cellular FLICE-like inhibitory protein; enhanced chemiluminescence; extracellular matrix; horseradish peroxidase; p-Akt; phosphate buffered saline; phosphoinositide 3-kinase; phosphor-Akt; serine/threonine protein kinase B; siRNA; small interfering RNA; sodium dodecyl sulfate