Abstracts Posters SICOT-SOF meeting Gothenburg 2010 _2_

Poster
Topic: Tumours
Abstract number: 25246
THE ROLE OF PI3K/AKT PATHWAY IN BONE AND SOFT TISSUE SARCOMAS
Yoshiki YAMAGAMI, Toshiaki HITORA, Osamu NAKAMURA, Tetsuji YAMAMOTO
Department of Orthopaedic Surgery, Kagawa University, kita-gun, kagawa (JAPAN)
INTRODUCTIONThe phosphatidylinositol 3-kinase (PI3K)/Akt pathway plays an
important role in various cellular processes including cell growth, survival, and
motility. We examined the expression of Akt and the existence of PI3K/Akt pathway
in human bone and soft tissue sarcoma cell lines.MATERIALS AND METHODSCell
lines and reagent: 3 human osteosarcoma cell lines and 3 human MFH cell lines
were used in this study. Akt inhibitor X, a specific Akt kinase inhibitor was used.The
inhibitory effect of Akt inhibitor: The cell proliferation was assayed using the MTS
assay. Cells were pretreated for 24 or 48h with Akt inhibitor. The percent viability of
each well was calculated.Western blotting: Cells were pretreated for 60 min with Akt
inhibitor, and immunoblotted with anti-AKT1/2/3 and anti-phospho-AKT1/2/3
antibody. Apoptosis Detection:ApoStrand ELISA Apoptosis Detection Kit which
detects single-stranded DNA was used. RESULTSThe effect of the Akt inhibitorAkt
inhibitor inhibited the cell proliferation of all cell lines. Expression of Akt and phospho-
AktNot only Akt but phospho- Akt were expressed in all cell lines. Phosphorylation of
Akt was decreased by Akt inhibitor in all cell lines.Apoptosis DetectionAn apoptotic
effect of Akt inhibitor was detected in all cell lines. DISCUSSIONIn our study, Akt
phosphorylation was detected and Akt inhibitor decreased the phosphorylation.
Moreover, Akt inhibitor inhibited cell proliferation and induced apoptosis. These
results suggest that PI3K/Akt signaling pathway exists and plays an important role in
bone and soft tissue sarcomas.
642