TY - JOUR
T1 - Absolute quantification of four isoforms of the class I phosphoinositide-3-kinase catalytic subunit by real-time RT-PCR
AU - Nakamura, Hiroyuki
AU - Dan, Shingo
AU - Akashi, Tetsuyuki
AU - Unno, Michiaki
AU - Yamori, Takao
N1 - Copyright:
Copyright 2009 Elsevier B.V., All rights reserved.
PY - 2007/6
Y1 - 2007/6
N2 - Class I phosphoinositide-3-kinase (PI3K) consists of four isoforms of the catalytic subunit, p110α, -β, -δ and -γ, generated from the genes PIK3CA, -B, -D and -G, respectively. These isoforms show different tissue distribution and some specific and indispensable functions in various biological pathways such as development, inflammation, autoimmunity and malignancy. In human cancers, frequent genomic amplification and gain-of-function mutations of PIK3CA were reported, which suggests an oncogenic potential. However, the role played by the other three isoforms in human cancer remains to be determined. We wanted to investigate the relationship between all the isoforms in human cancers. Here, we have established a system for the simultaneous absolute-quantification of all four isoforms by real-time reverse transcription polymerase chain reaction (RT-PCR). The reliability of this system was confirmed using three main criteria: (i) good correlation of each standard curve, (ii) high specificity of the PCR reactions and (iii) excellent reproducibility. Using this system, we investigated human monocytic leukemia cells (U937) to analyze expression of all four isoforms. The biological implications of the expression level of the four isoforms of class I PI3K catalytic subunit are discussed.
AB - Class I phosphoinositide-3-kinase (PI3K) consists of four isoforms of the catalytic subunit, p110α, -β, -δ and -γ, generated from the genes PIK3CA, -B, -D and -G, respectively. These isoforms show different tissue distribution and some specific and indispensable functions in various biological pathways such as development, inflammation, autoimmunity and malignancy. In human cancers, frequent genomic amplification and gain-of-function mutations of PIK3CA were reported, which suggests an oncogenic potential. However, the role played by the other three isoforms in human cancer remains to be determined. We wanted to investigate the relationship between all the isoforms in human cancers. Here, we have established a system for the simultaneous absolute-quantification of all four isoforms by real-time reverse transcription polymerase chain reaction (RT-PCR). The reliability of this system was confirmed using three main criteria: (i) good correlation of each standard curve, (ii) high specificity of the PCR reactions and (iii) excellent reproducibility. Using this system, we investigated human monocytic leukemia cells (U937) to analyze expression of all four isoforms. The biological implications of the expression level of the four isoforms of class I PI3K catalytic subunit are discussed.
KW - Absolute quantification
KW - Phosphoinositide-3-kinase (PI3K)
KW - Real-time reverse transcription polymerase chain reaction (RT-PCR)
KW - U937
UR - http://www.scopus.com/inward/record.url?scp=34249998437&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=34249998437&partnerID=8YFLogxK
U2 - 10.1248/bpb.30.1181
DO - 10.1248/bpb.30.1181
M3 - Article
C2 - 17541179
AN - SCOPUS:34249998437
VL - 30
SP - 1181
EP - 1184
JO - Biological and Pharmaceutical Bulletin
JF - Biological and Pharmaceutical Bulletin
SN - 0918-6158
IS - 6
ER -