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Protein-tyrosine kinase and protein-serine/threonine kinase expression in human gastric cancer cell lines
Authors:Jyh-Shi Lin  Chi-Wei Lu  Chang-Jen Huang  Peng-Fyn Wu  Daniel Robinson  Hsing-Jien Kung  Chin-Wen Chi  Chew-Wun Wu  Wen-Kang Yang  Jacqueline J. K. Whang-Peng  Wen-chang Lin PhD
Affiliation:(1) Institute of Biomedical Sciences, Academia Sinica and Clinical Cancer Center, National Health Research Institutes, Taipei, Taiwan, ROC;(2) Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan, ROC;(3) Department of Molecular Biology and Microbiology, School of Medicine, Case Western Reserve University, Cleveland, Ohio, USA;(4) Department of Surgery, National Yang-Ming University, Taipei, Taiwan, ROC;(5) Department of Medical Research, Veterans General Hospital-Taipei and School of Medicine, National Yang-Ming University, Taipei, Taiwan, ROC
Abstract: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.
Keywords:Protein tyrosine kinase  Protein serine/threonine kinase  RT-PCR  Gastric cancer
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