Methods in enzymology
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Methods in enzymology · Jan 2010
Large-scale glycomics for discovering cancer-associated N-glycans by integrating glycoblotting and mass spectrometry.
It has known that the glycosylation plays an important role in the biological states, such as development, aging, and diseases. Although genomic and proteomic approaches have been intensively studied for diagnosis and disease treatment, glycomics have been laggard compared to them due to the hardness of the purification procedure from crude biological materials. Recently, we have developed "glycoblotting" method, a high-throughput and quantitative technique for comprehensive glycomics, which enables to enrich and quantify glycans from crude biological materials, such as serum, tissue biopsy, and cell lysate [Niikura, K., Kamitani, R., Kurogochi, M., Uematsu, R., Shinohara, Y., Nakagawa, H., Deguchi, K., Monde, K., Kondo, H., and Nishimuram S.-I. (2005). ⋯ Cell. Proteomics9, 523-537]. In this chapter, we will discuss glycoblotting method including the potentials not only for exploration of glycan-related cancer biomarker but also for detection of cellular differentiation.
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Methods in enzymology · Jan 2008
Chapter 11. The role of bone marrow-derived cells in tumor angiogenesis and metastatic progression.
Tumor angiogenesis is orchestrated by a complex set of secreted factors and collaboration between many different cell types. This chapter discusses the role of tumor-secreted angiogenic factors that are responsible for the recruitment of bone marrow-derived cells (BMDCs) at tumor sites and their role in tumor angiogenesis and progression.
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In addition to their contribution to research fields such as early human development, self-renewal, and differentiation mechanisms, human embryonic stem cells (hESCs) may serve as a tool for drug testing and for the study of cell-based therapies. Traditionally, these cells have been cultured with mouse embryonic fibroblast (MEF) feeder layers, which allow their continuous growth in an undifferentiated state. ⋯ To this end, different culture methods for hESCs, based on serum replacement and free of supportive cell layers, were developed. This chapter discusses a simple, feeder-free culture system on the basis of medium supplemented with transforming growth factor beta1 (TGFbeta1), basic fibroblast growth factor (bFGF) and fibronectin as matrix.
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Methods in enzymology · Jan 2006
Sorafenib (BAY 43-9006, Nexavar), a dual-action inhibitor that targets RAF/MEK/ERK pathway in tumor cells and tyrosine kinases VEGFR/PDGFR in tumor vasculature.
Activating mutations in Ras and B-RAF were identified in several human cancers. In addition, several receptor tyrosine kinases, acting upstream of Ras, were found either mutated or overexpressed in human tumors. Because oncogenic activation of the Ras/RAF pathway may lead to a sustained proliferative signal resulting in tumor growth and progression, inhibition of this pathway represents an attractive approach for cancer drug discovery. ⋯ Thus, sorafenib may inhibit tumor growth by a dual mechanism, acting either directly on the tumor (through inhibition of Raf and Kit signaling) and/or on tumor angiogenesis (through inhibition of VEGFR and PDGFR signaling). In phase I and phase II clinical trials, sorafenib showed limited side effects and, more importantly, disease stabilization. This agent is currently being evaluated in phase III clinical trials in renal cell and hepatocellular carcinomas.
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Repair or regeneration of defective lung epithelium would be of great therapeutic potential. Cellular sources for such repair have long been searched for within the lung, but the identification and characterization of stem or progenitor cells have been hampered by the complexity and cellular heterogeneity of the organ. In recent years, various pulmonary cells have been identified that meet the criteria for stem cells but it remains to be seen how far manipulation of these tissue-specific cell pools can upregulate epithelial repair. ⋯ The potential of human embryonic stem cells to generate any cell, tissue, or organ on demand for tissue repair or replacement is promising to revolutionize the treatment of human disease. Although some headway has been made into making pulmonary epithelium from these stem cells, human embryonic stem cell technology is still in its infancy and many technical, safety, and ethical hurdles must be cleared before clinical trials can begin. This chapter focuses on the potential role of stem cells in future approaches to lung repair and regeneration.