Protein-tyrosine kinase and protein-serine/threonine kinase expression in human gastric cancer cell lines
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Protein kinases play key roles in cellular functions. They are involved in many cellular functions including; signal transduction, cell cycle regulation, cell division, and cell differentiation. Alterations of protein kinase by gene amplification, mutation or viral factors often induce tumor formation and tumor progression toward malignancy. The identification and cloning of kinase genes can provide a better understanding of the mechanisms of tumorigenesis as well as diagnostic tools for tumor staging. In this study, we have used degenerated polymerase-chain-reaction primers according to the consensus catalytic domain motifs to amplify protein kinase genes (protein-tyrosine kinase, PTK, and protein-serine/threonine kinase, PSK) from human stomach cancer cells. Following amplification, the protein kinase molecules expressed in the gastric cancer cells were cloned into plasmid vectors for cloning and sequencing. Sequence analysis of polymerase-chain-reaction products resulted in the identification of 25 protein kinases, including two novel ones. Expression of several relevant PTK/PSK genes in gastric cancer cells and tissues was further substantiated by RT-PCR using gene-specific primers. The identification of protein kinases expressed or activated in the gastric cancer cells provide the framework to understand the oncogenic process of stomach cancer.
Key WordsProtein tyrosine kinase Protein serine/threonine kinase RT-PCR Gastric cancer
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- 6.Hanks SK, Hunter T. The eukaryotic protein kinase superfamily. In: Hardie G, Hanks S, eds. The Protein Kinase Facts Book. San Diego, Academic Press, I:7–47;1995.Google Scholar
- 7.Hanks SK, Quinn AM, Hunter T. The protein kinase family conserved features and deduced phylogeny of the catalytic domains. Science 241:42–52;1988.Google Scholar
- 14.Jin LD, Meng CL, Han SH, Ding MJ, Chang TM, Chan TK, Shen KL. A study on production of monoclonal antibodies using SC-M1 cells as immunogen. Med Sci 8:17–25;1987.Google Scholar
- 17.Maniatis T, Fritsch E, Sambrook J. Molecular Cloning: A Laboratory Manual. ed 1. Cold Spring Harbor, Cold Spring Harbor Laboratory, 1982.Google Scholar
- 20.Schultz SJ, Nigg EA. Identification of 21 novel human protein kinases, including 3 members of a family related to the cell cycle regulator nima ofAspergillus nidulans. Cell Growth Different 4:821–830;1993.Google Scholar
- 22.Shu HK, Pelley RJ, Kung HJ. Tissue-specific transformation by epidermal growth factor receptor: A single point mutation within the ATP-binding pocket of the erbB product increases its intrinsic kinase activity and activates its sarcomagenic potential. Proc Natl Acad Sci USA 87:9103–9107;1990.PubMedGoogle Scholar
- 23.Takahashi T, Shirasawa T, Miyake K, Yahagi Y, Maruyama N, Kasahara N, Kawamura T, Matsumura O, Mitarai T, Sakai O. Protein tyrosine kinases expressed in glomeruli and cultured glomerular cells: FLT-1 and VEGF expression in renal mesangial cells. Biochem Biophys Res Commun 209:218–226;1995.CrossRefPubMedGoogle Scholar
- 29.Yokota J, Yamamoto T, Miyajima N, Toyoshima K, Nomura N, Sakamoto H, Yoshida T, Terada M, Sugimura T. Genetic alterations of the c-erbB-2 oncogene occur frequently in tubular adenocarcinoma of the stomach and are often accompanied by amplification of the v-erb A homologue. Oncogene 2:283–287;1988.PubMedGoogle Scholar