دورية أكاديمية
Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking.
| العنوان: | Subcellular Fractionation of Primary Chronic Lymphocytic Leukemia Cells to Monitor Nuclear/Cytoplasmic Protein Trafficking. |
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| المؤلفون: | Hay J; Paul O'Gorman Leukaemia Research Centre, Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow., Moles MW; Paul O'Gorman Leukaemia Research Centre, Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow., Cassels J; Paul O'Gorman Leukaemia Research Centre, Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow., Michie AM; Paul O'Gorman Leukaemia Research Centre, Institute of Cancer Sciences, College of Medical, Veterinary and Life Sciences, University of Glasgow; Alison.Michie@glasgow.ac.uk. |
| المصدر: | Journal of visualized experiments : JoVE [J Vis Exp] 2019 Oct 23 (152). Date of Electronic Publication: 2019 Oct 23. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't; Video-Audio Media |
| اللغة: | English |
| بيانات الدورية: | Publisher: MYJoVE Corporation Country of Publication: United States NLM ID: 101313252 Publication Model: Electronic Cited Medium: Internet ISSN: 1940-087X (Electronic) Linking ISSN: 1940087X NLM ISO Abbreviation: J Vis Exp Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: [Boston, Mass. : MYJoVE Corporation, 2006]- |
| مواضيع طبية MeSH: | Cell Fractionation/*methods , Leukemia, Lymphocytic, Chronic, B-Cell/*metabolism , Protein Transport/*physiology, Active Transport, Cell Nucleus ; Carrier Proteins/metabolism ; Cell Nucleus/metabolism ; Cytoplasm/metabolism ; Cytosol/metabolism ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell/pathology ; Nuclear Proteins/metabolism ; Signal Transduction |
| مستخلص: | Nuclear export of macromolecules is often deregulated in cancer cells. Tumor suppressor proteins, such as p53, can be rendered inactive due to aberrant cellular localization disrupting their mechanism of action. The survival of chronic lymphocytic leukaemia (CLL) cells, among other cancer cells, is assisted by the deregulation of nuclear to cytoplasmic shuttling, at least in part through deregulation of the transport receptor XPO1 and the constitutive activation of PI3K-mediated signaling pathways. It is essential to understand the role of individual proteins in the context of their intracellular location to gain a deeper understanding of the role of such proteins in the pathobiology of the disease. Furthermore, identifying processes that underlie cell stimulation and the mechanism of action of specific pharmacological inhibitors, in the context of subcellular protein trafficking, will provide a more comprehensive understanding of the mechanism of action. The protocol described here enables the optimization and subsequent efficient generation of nuclear and cytoplasmic fractions from primary chronic lymphocytic leukemia cells. These fractions can be used to determine changes in protein trafficking between the nuclear and cytoplasmic fractions upon cell stimulation and drug treatment. The data can be quantified and presented in parallel with immunofluorescent images, thus providing robust and quantifiable data. |
| معلومات مُعتمدة: | MR/K014854/1 United Kingdom MRC_ Medical Research Council |
| المشرفين على المادة: | 0 (Carrier Proteins) 0 (Nuclear Proteins) |
| تواريخ الأحداث: | Date Created: 20191112 Date Completed: 20200625 Latest Revision: 20210304 |
| رمز التحديث: | 20231215 |
| DOI: | 10.3791/60426 |
| PMID: | 31710039 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-087X |
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| DOI: | 10.3791/60426 |