Abstract
Phosphatidylinositol-4-phosphate 5-kinase-like 1 (PIP5KL1), the forth member of phosphatidylinositol-4-phosphate 5-kinases (PIPKs) type I, acts as a scaffold for localization and activation of PIPKs, which mediates numerous cellular processes. However, the role of PIP5KL1 in the development of human cancer is still lacking. We therefore examined the expression of PIP5KL1 in human normal and cancer tissues by tissue microarrays (TMAs). Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence imaging analysis were used to testify the mRNA and protein levels of PIP5KL1 in human gastric cancer cell line (BGC823). The cell proliferation was investigated with 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) assay. Both wound healing and transwell migration assay were performed to study the cell migration. The phosphorylation of v-akt murine thymoma viral oncogene homolog 1 (AKT1) was determined by western immunoblot analysis. Immunostaining of gastric cancer tissue microarrays revealed a negative correlation between PIP5KL1 overexpression and gastric cancer in situ. Transient transfection PIP5KL1 induced a significant increase expression at both transcriptional and translational levels and consequent robust inhibition of proliferation (P < 0.05) and migration (P < 0.05) of BGC823 cells. Overexpression of PIP5KL1 markedly inhibited (P < 0.05) serum-induced phosphorylation of AKT1. Taken together, these studies indicate a functional negative correlation between elevated levels of PIP5KL1 and the development of human gastric cancer, suggesting that PIP5KL1 overexpression may suppress gastric cancer formation.
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References
Toker A (2002) Phosphoinositides and signal transduction. Cell Mol Life Sci 59:761–779
Gardocki ME, Jani N, Lopes JM (2005) Phosphatidylinositol biosynthesis: biochemistry and regulation. Biochim Biophys Acta 1735:89–100
Blero D, Payrastre B, Schurmans S, Erneux C (2007) Phosphoinositide phosphatases in a network of signalling reactions. Pflugers Arch 455:31–44
Wymann MP, Schneiter R (2008) Lipid signalling in disease. Nat Rev Mol Cell Biol 9:162–176
Pendaries C, Tronchère H, Plantavid M, Payrastre B (2003) Phosphoinositide signaling disorders in human diseases. FEBS Lett 546:25–31
Stace C, Manifava M, Delon C, Coadwell J, Cockcroft S, Ktistakis NT (2008) PA binding of phosphatidylinositol 4-phosphate 5-kinase. Adv Enzyme Regul 48:55–72
Loijens JC, Boronenkov IV, Parker GJ, Anderson RA (1996) The phosphatidylinositol 4-phosphate 5-kinase family. Adv Enzyme Regul 36:115–140
Chang JD, Field SJ, Rameh LE, Carpenter CL, Cantley LC (2004) Identification and characterization of a phosphoinositide phosphate kinase homolog. J Biol Chem 279:11672–11679
Wang L, Gao X, Gao P, Deng W, Yu P, Ma J, Guo J, Wang X, Cheng H, Zhang C, Yu C, Ma X, Lv B, Lu Y, Shi T, Ma D (2006) Cell-based screening and validation of human novel genes associated with cell viability. J Biomol Screen 11:369–376
Luoh SW, Venkatesan N, Tripathi R (2004) Overexpression of the amplified Pip4k2beta gene from 17q11–12 in breast cancer cells confers proliferation advantage. Oncogene 23:1354–1363
Akiyama C, Shinozaki-Narikawa N, Kitazawa T, Hamakubo T, Kodama T, Shibasaki Y (2005) Phosphatidylinositol-4-phosphate 5-kinase gamma is associated with cell-cell junction in A431 epithelial cells. Cell Biol Int 29:514–520
Do GM, Choi MS, Kim HJ, Woo MN, Lee MK, Jeon SM (2008) Soy pinitol acts partly as an insulin sensitizer or insulin mediator in 3T3–L1 preadipocytes. Genes Nutr 2:359–364
Yarrow JC, Perlman ZE, Westwood NJ, Mitchison TJ (2004) A high-throughput cell migration assay using scratch wound healing, a comparison of image-based readout methods. BMC Biotechnol 4:21–30
Voduc D, Kenney C, Nielsen TO (2008) Tissue microarrays in clinical oncology. Semin Radiat Oncol 18:89–97
Verderio P, Carbone A (2009) Tissue microarrays for immunohistochemical determination of oncological biomarkers. Virchows Arch 454:353–354
Heck JN, Mellman DL, Ling K, Sun Y, Wagoner MP, Schill NJ, Anderson RA (2007) A conspicuous connection: structure defines function for the phosphatidylinositol-phosphate kinase family. Crit Rev Biochem Mol Biol 42:15–39
Honda A, Nogami M, Yokozeki T, Yamazaki M, Nakamura H, Watanabe H, Kawamoto K, Nakayama K, Morris AJ, Frohman MA, Kanaho Y (1999) Phosphatidylinositol 4-phosphate 5-kinase alpha is a downstream effector of the small G protein ARF6 in membrane ruffle formation. Cell 99:521–532
Coppolino MG, Dierckman R, Loijens J, Collins RF, Pouladi M, Jongstra-Bilen J, Schreiber AD, Trimble WS, Anderson R, Grinstein S (2002) Inhibition of phosphatidylinositol-4-phosphate 5-kinase I alpha impairs localized actin remodeling and suppresses phagocytosis. J Biol Chem 277:43849–43857
Yamamoto M, Hilgemann DH, Feng S, Bito H, Ishihara H, Shibasaki Y, Yin HL (2001) Phosphatidylinositol 4,5-bisphosphate induces actin stress-fiber formation and inhibits membrane ruffling in CV1 cells. J Cell Biol 152:867–876
Ling K, Doughman RL, Firestone AJ, Bunce MW, Anderson RA (2002) Type I gamma phosphatidylinositol phosphate kinase targets and regulates focal adhesions. Nature 420:89–93
Kisseleva M, Feng Y, Ward M, Song C, Anderson RA, Longmore GD (2005) The LIM protein Ajuba regulates phosphatidylinositol 4,5-bisphosphate levels in migrating cells through an interaction with and activation of PIPKI alpha. Mol Cell Biol 25:3956–3966
Chandrasekar I, Stradal TE, Holt MR, Entschladen F, Jockusch BM, Ziegler WH (2005) Vinculin acts as a sensor in lipid regulation of adhesion-site turnover. J Cell Sci 118(Pt 7):1461–1472
Doughman RL, Firestone AJ, Anderson RA (2003) Phosphatidylinositol phosphate kinases put PI4, 5P(2) in its place. J Membr Biol 194:77–89
Boronenkov IV, Loijens JC, Umeda M, Anderson RA (1998) Phosphoinositide signaling pathways in nuclei are associated with nuclear speckles containing pre-mRNA processing factors. Mol Biol Cell 9:3547–3560
Di Paolo G, Pellegrini L, Letinic K, Cestra G, Zoncu R, Voronov S, Chang S, Guo J, Wenk MR, De Camilli P (2002) Recruitment and regulation of phosphatidylinositol phosphate kinase type 1 gamma by the FERM domain of talin. Nature 420:85–89
Somanath PR, Razorenova OV, Chen J, Byzova TV (2006) Akt1 in endothelial cell and angiogenesis. Cell Cycle 5:512–518
Peng SB, Peek V, Zhai Y, Paul DC, Lou Q, Xia X, Eessalu T, Kohn W, Tang S (2005) Akt activation, but not extracellular signal-regulated kinase activation, is required for SDF-1alpha/CXCR4-mediated migration of epithelioid carcinoma cells. Mol Cancer Res 3:227–236
Toker A, Yoeli-Lerner M (2006) Akt signaling and cancer: surviving but not moving on. Cancer Res 66:3963–3966
Manning BD, Cantley LC (2007) AKT/PKB signaling: navigating downstream. Cell 129:1261–1274
Mejillano M, Yamamoto M, Rozelle AL, Sun HQ, Wang X, Yin HL (2001) Regulation of apoptosis by phosphatidylinositol 4, 5-bisphosphate inhibition of caspases, and caspase inactivation of phosphatidylinositol phosphate 5-kinases. J Biol Chem 276:1865–1872
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Shi, L., Zhao, M., Luo, Q. et al. Overexpression of PIP5KL1 suppresses cell proliferation and migration in human gastric cancer cells. Mol Biol Rep 37, 2189–2198 (2010). https://doi.org/10.1007/s11033-009-9701-5
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DOI: https://doi.org/10.1007/s11033-009-9701-5