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Localization of Prohibitin in the Nuclear Matrix and Alteration of Its Expression During Differentiation of Human Neuroblastoma SK-N-SH Cells Induced by Retinoic Acid
Authors:Qi-Fu Li  Ying Liang  Song-Lin Shi  Qing-Rong Liu  Dong-Hui Xu  Guang-Jun Jing  San-Ying Wang  Hai-Yan Kong
Institution:(1) The Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Science, Xiamen University, Xiamen, 361005, China;(2) Medical College of Xiamen University, Xiamen, 361005, China;(3) Department of Health and Human Services (DHSS), Molecular Neurobiology Branch, National Institute on Drug Abuse-Intramural Research Program (NIDA-IRP), NIH, 333 Cassell Drive, Baltimore, MD 21224, USA;(4) The Affiliated First Hospital of Medical College, Xiamen University, Xiamen, 361005, China
Abstract:The nuclear matrix-intermediate filament system of human neuroblastoma SK-N-SH cells before and after retinoic acid (RA) treatment was selectively extracted and the distribution of prohibitin (PHB) in the nuclear matrix, as well as its colocalization with related genes, was observed. Results of two-dimensional gel electrophoresis (2-DE), mass spectrometry (MS) identification, and protein immunoblotting all confirm that PHB was present in the components of SK-N-SH nuclear matrix proteins and was down-regulated after RA treatment. Immunofluorescence microscopy observations show that PHB was localized in the nuclear matrix and its distribution was altered due to RA treatment. Laser confocal microscopy results reveal that PHB colocalized with the expression products of c-myc, c-fos, p53, and Rb, but the colocalization region was altered after RA treatment. Our results prove that PHB is a nuclear matrix protein and is localized in nuclear matrix fibers. The distribution of PHB in SK-N-SH cells and its colocalization with related proto-oncogenes and tumor suppressor genes suggest that PHB plays pivotal roles in the differentiation of SK-N-SH cells and deserves further study.
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