BACKGROUND: Class IA PI 3-kinases produce phosphatidylinositol (3,4,5)-trisphosphate (PIP3). PIP3 is bound by AKT which facilities its activation by PDK1. Activated AKT promotes cell survival and stimulates cell proliferation. Class IA PI 3-kinases are heterodimers consisting of a regulatory subunit p85 and a catalytic subunit p110. The p110alpha isoform has been shown to be mutated in a number of tumor types. A number of recent studies suggest that the p110beta isoform may be functionally relevant in prostate cancer. In this study we extend this work to include the examination of the expression and functional properties of p110alpha and p110beta in three different prostate cancer cell lines, DU145, LNCaP, PC3, as well as the non-tumorigenic but immortalized RWPE1 prostate epithelial cell line. METHODS: Western blot analysis was used to measure protein expression and quantitative real-time PCR was used to measure mRNA levels. After targeted knockdown using isoform-specific siRNAs to reduce PI 3-kinase p110alpha or p110beta isoform expression, we measured downstream signally events such as phosphorylation of AKT, ERK 1/2, PDK, and FOXO, as well as biological consequences such as changes in apoptosis, and alterations in cell cycle progression. RESULTS: In all three prostate cancer cell lines examined, targeted knockdown of p110beta, and not p110alpha, resulted in significantly reduced AKT, PDK, and FOXO phosphorylation. While knockdown of either p110 isoform resulted in an increase in apoptosis and a cell cycle arrest in G1 in the remaining non-apoptotic cells, these effects were much more pronounced with knockdown of p110beta. CONCLUSIONS: Our results support the concept that p110beta appears to be the predominant functional class I PI 3-kinase isoform in prostate cancer cells.
BACKGROUND: Class IA PI 3-kinases produce phosphatidylinositol (3,4,5)-trisphosphate (PIP3). PIP3 is bound by AKT which facilities its activation by PDK1. Activated AKT promotes cell survival and stimulates cell proliferation. Class IA PI 3-kinases are heterodimers consisting of a regulatory subunit p85 and a catalytic subunit p110. The p110alpha isoform has been shown to be mutated in a number of tumor types. A number of recent studies suggest that the p110beta isoform may be functionally relevant in prostate cancer. In this study we extend this work to include the examination of the expression and functional properties of p110alpha and p110beta in three different prostate cancer cell lines, DU145, LNCaP, PC3, as well as the non-tumorigenic but immortalized RWPE1 prostate epithelial cell line. METHODS: Western blot analysis was used to measure protein expression and quantitative real-time PCR was used to measure mRNA levels. After targeted knockdown using isoform-specific siRNAs to reduce PI 3-kinase p110alpha or p110beta isoform expression, we measured downstream signally events such as phosphorylation of AKT, ERK 1/2, PDK, and FOXO, as well as biological consequences such as changes in apoptosis, and alterations in cell cycle progression. RESULTS: In all three prostate cancer cell lines examined, targeted knockdown of p110beta, and not p110alpha, resulted in significantly reduced AKT, PDK, and FOXO phosphorylation. While knockdown of either p110 isoform resulted in an increase in apoptosis and a cell cycle arrest in G1 in the remaining non-apoptotic cells, these effects were much more pronounced with knockdown of p110beta. CONCLUSIONS: Our results support the concept that p110beta appears to be the predominant functional class I PI 3-kinase isoform in prostate cancer cells.
Authors: Hong Lin; Mark J Schulz; Ren Xie; Jin Zeng; Juan I Luengo; Michael D Squire; Rosanna Tedesco; Junya Qu; Karl Erhard; James F Mack; Kaushik Raha; Ramona Plant; Cynthia M Rominger; Jennifer L Ariazi; Christian S Sherk; Michael D Schaber; Jeanelle McSurdy-Freed; Michael D Spengler; Charles B Davis; Mary Ann Hardwicke; Ralph A Rivero Journal: ACS Med Chem Lett Date: 2012-05-29 Impact factor: 4.345
Authors: Hongyi Yu; Michael L Moore; Karl Erhard; Mary Ann Hardwicke; Hong Lin; Juan I Luengo; Jeanelle McSurdy-Freed; Ramona Plant; Junya Qu; Kaushik Raha; Cynthia M Rominger; Michael D Schaber; Michael D Spengler; Ralph A Rivero Journal: ACS Med Chem Lett Date: 2013-01-10 Impact factor: 4.345
Authors: Pierre-Olivier Estève; Yanqi Chang; Mala Samaranayake; Anup K Upadhyay; John R Horton; George R Feehery; Xiaodong Cheng; Sriharsa Pradhan Journal: Nat Struct Mol Biol Date: 2010-12-12 Impact factor: 15.369