Comparative proteomic analyses on the differentiation of dimethyl sulfoxide induced murine erythroleukemia cells

YANG Zu-li  ZHU Xiao-fang  SHI Ming-ming  HU Fan  ZHAO Fu-kun  ZHANG Shi-fu*

doi:10.3969/j.issn.0529-1356.2014.04.012

Acta Anatomica Sinica ›› 2014, Vol. 45 ›› Issue (4) : 507-515. DOI: 10.3969/j.issn.0529-1356.2014.04.012

Comparative proteomic analyses on the differentiation of dimethyl sulfoxide induced murine erythroleukemia cells

YANG Zu-li ZHU Xiao-fang SHI Ming-ming HU Fan ZHAO Fu-kun ZHANG Shi-fu*

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Abstract

Objective To explore the differentially expressed proteins during erythroid differentiation. Methods The murine erythroleukemia (MEL) cell were treated by DMSO, and the comparative proteomic was systematically analyzed and identified on different differentiating time points. ratio of cell differentiation and viability were detected by benzidine staining, MTT assay and Ter119 immunofluorescence. Using two-dimensional gel electrophoresis combined with mass spectrometry technology and bioinformatics analysis, we conducted a comparative proteomic analysis on MEL cells during the process of induced differentiation to screen and identify differential proteins. Results The MEL cells induced by 1.2% DMSO for 0 hour, 6hours, 12hours, 24hours, 36hours, 48hours, 72 hours, 96 hours, 120 hours were collected for proteomic analysis, by two-dimensional gel electrophoresis combined with mass spectrometry. A total of 87 kinds of proteins were successfully identified. MEL cells exposed to DMSO at a final concentration of 1.2% for 120 hours reached the highest differentiation rate of 67%. MTT assay showed that 1.0%, 1.2%, 1.4% DMSO had no inhibiting effect on cell vitality. Conclusion DMSO may induce MEL cells to differentiate and have no inhibiting effect on cell vitality. The 87 kinds of differentially expressed proteins from two-dimentional gel electrophoresis may be divided into twelve categories; the most three parts are 41% enzyme protein, 15% structural protein and 13% regulatory protein.

Key words

Murine erythroleukemia cell / Dimethyl sulfoxide / Induce / Differentiation / Two-dimentional gel electrophoresis / Mouse

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YANG Zu-li ZHU Xiao-fang SHI Ming-ming HU Fan ZHAO Fu-kun ZHANG Shi-fu*. Comparative proteomic analyses on the differentiation of dimethyl sulfoxide induced murine erythroleukemia cells[J]. Acta Anatomica Sinica. 2014, 45(4): 507-515 https://doi.org/10.3969/j.issn.0529-1356.2014.04.012

References

［1］Marks P, Rifkind R. Erythroleukemic differentiation［J］. Annu Rev Biochem, 1978, 47(1): 419-448.
［2］Moreau-Gachelin F. Multi-stage friend murine erythroleukemia: Molecular insights into oncogenic cooperation［J］. Retrovirology, 2008, 5(1): 99.
［3］Lee CR, Cervi D, Truong AH, et al. Friend virus-induced erythroleukemias: a unique and well-defined mouse model for the development of leukemia［J］. Anticancer Res, 2003, 23(3A): 2159-2166.
［4］Friend C, Scher W, Holland JG, et al. Hemoglobin synthesis in murine virus-induced leukemic cells in vitro: stimulation of erythroid differentiation by dimethyl sulfoxide［J］. Proc Natl Acad Sci USA, 1971, 68(2): 378-382.
［5］Tanaka M, Levy J, Terada M, et al. Induction of erythroid differentiation in murine virus infected eythroleukemia cells byhighly polar compounds［J］. Proc Natl Acad Sci USA, 1975, 72(3): 1003-1006.
［6］Holbrook I, Leaver A. A procedure to increase the sensitivity of staining by coomassie brilliant blue G250-perchloric acid solution［J］. Anal Biochem, 1976, 75(2): 634-636.
［7］Wray W, Boulikas T, Wray VP, et al. Silver staining of proteins in polyacrylamide gels［J］. Anal Biochem, 1981, 118(1): 197-203.
［8］Zhao Q, Ge Y, Zhou J, et al. Differentiation and malignant suppression induced by mouse erythroid differentiation and denucleation factor on mouse erythroleukemia cells［J］. Zhongguo Yi Xue Ke Xue Yuan Xue Bao, 2001, 23(1): 32-35.
［9］Koury MJ, Sawyer ST, Brandt SJ. New insights into erythropoiesis［J］. Curr Opin Hematol, 2002, 9(2): 93-100.
［10］Zuo Z, Polski J M, Kasyan A, et al. Acute erythroid leukemia［J］. Arch Pathol Lab Med, 2010, 134(9): 1261-1270.
［11］Müller WE, Qiang L, Schrder HC, et al. Carbonic anhydrase: a key regulatory and detoxifying enzyme for karst plants［J］. Planta, 2014, 239(1): 213-229.
［12］Ferreira JC, Icimoto MY, Marcondes MF, et al. Recycling of the high valence states of heme proteins by cysteine residues of THIMET-oligopeptidase［J］. PLoS One, 2013, 8(11): e79102.
［13］Saarikangas J, Barral Y. The emerging functions of septins in metazoans［J］. EMBO Rep, 2011, 12(11): 1118-1126.
［14］Prag S, Parsons M, Keppler MD, et al. Activated ezrin promotes cell migration through recruitment of the gef dbl to lipid rafts and preferential downstream activation of cdc42［J］. Mol Biol Cell, 2007, 18(8): 2935-2948.
［15］Yu Y, Khan J, Khanna C, et al. Expression profiling identifies the cytoskeletal organizer ezrin and the developmental homeoprotein six-1 as key metastatic regulators［J］. Nat Med, 2004, 10(2): 175-181.
［16］Tsai CL, Tsai CN, Lin CY, et al. Secreted stress-induced phosphoprotein 1 activates the alk2-smad signaling pathways and promotes cell proliferation of ovarian cancer cells［J］. Cell Rep, 2012, 2(2): 283-293.
［17］Chao A, Lee LY, Hsueh C, et al. Immunohistological analysis of stress-induced phosphoprotein 1 in ovarian cancer patients with low serum cancer antigen 125 levels［J］. Taiwan J Obstet Gynecol, 2013, 52(2): 185-191.
［18］Yawata M, Murata S, Tanaka K, et al. Nucleotide sequence analysis of the approximately 35-kb segment containing interferon-gamma-inducible mouse proteasome activator genes［J］. Immunogenetics, 2001, 53(2): 119-129.