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p38-2G4蛋白高表达对小鼠红白血病细胞增殖和分化的影响
Effects of p38-2G4 high-expression on the proliferation and erythriod differentiateon of murine erythroleukemia cells
目的 探讨小鼠增殖相关蛋白p38-2G4基因表达上调对小鼠红白血病（MEL）细胞增殖和诱导分化能力的影响。方法 构建p38-2G4-pLJM1高表达重组载体与pCMV-VSV-G和pCMV-dR8.2包装质粒共转染HEK293T细胞慢病毒，感染MEL细胞，建立稳定高表达p38-2G4蛋白的MEL细胞系，Western blotting检测稳定株p38-2G4蛋白的表达。MTT实验和联苯胺染色法检测p38-2G4蛋白高表达对MEL细胞增殖和诱导分化能力的影响。 结果 高表达稳定株中p38-2G4蛋白表达量明显高于对照组细胞（P<0.05）。p38-2G4蛋白的高表达能显著影响MEL诱导分化过程，导致血红蛋白合成减少（P<0.05），但不能明显改变细胞活力（P>0.05）。结论 p38-2G4蛋白的高表达对MEL细胞增殖无明显影响，但能显著影响其被丁酸钠诱导分化的过程。
Objective To explore the effect of mouse proliferation-associated protein 2G4 (p38-2G4) high-expression on the proliferation and erythriod differentiation of murine erythroleukemia (MEL) cells. Methods To establish the recombinant lentivirus vector p38-2G4-pLJM1, the p38-2G4-pLJM1 was cotransfected into HEK293T cells to obtain lentivirus with pCMV-VSV-G and pCMV-dR8.2. Lentivirus were infected into MEL cells to establish the stably p38-2G4 high-expressed MEL cells. Western blotting was used to analyse the high-expression efficiency. MTT assay and benzidine staining were applied to detect the cell viability and hemoglobin synthesis of the stable cell line in presence/absence of inducers. Results Western blotting showed that the p38-2G4 high-expression stable cell strain had a higher expression of p38-2G4 as compared to the control group (MEL)（P<0.05）. MTT result showed that there was no difference between the p38-2G4 high-expression cell strain and the control group（P>0.05）, but the hemoglobin synthesis had been reduced as compared to the control group（P<0.05）. Conclusion p38-2G4 high-expression does not affect the cell viability of MEL cells, but inhibits the erythriod differentiation of MEL cells in three independent experiments.
p38-2G4 / 小鼠红白血病 / 慢病毒转染 / 高表达 / 免疫印迹法 / 小鼠
p38-2G4 / Murine erythroleukemia / Lentiviral transfection / High-expression / Western blotling / Mouse
[1］Radomski N, Jost E. Molecular cloning of a murine cdna encoding a novel protein, p38-2G4, which varies with the cell cycle［J］. Exp Cell Res, 1995, 220(2): 434-445.
［2］Zhang Y, Woodford N, Xia X, et al. Repression of E2F1-mediated transcription by the ErbB3 binding protein Ebp1 involves histone deacetylases［J］. Nucleic Acids Res, 2003, 31(8): 2168-2177.
［3］Zhang Y, Wang XW, Jelovac D, et al. The ErbB3-binding protein Ebp1 suppresses androgen receptor-mediated gene transcription and tumorigenesis of prostate cancer cells［J］. Proc Natl Acad Sci USA, 2005, 102(28): 9890-9895.
［4］Kim CK, Nguyen TL, Joo K M, et al. Negative regulation of p53 by the long isoform of erbb3 binding protein Ebp1 in brain tumors［J］. Cancer Res, 2010, 70(23): 9730-9741.
［5］Luo YX, Sun J, Yu YC. ［Expression and clinical significance of ErbB3 binding protein ebp1, E-cadherin, ICAM-1 and matrix metalloproteinase-9 in salivary adenoid cystic carcinoma］［J］. Shanghai Kou Qiang Yi Xue, 2013, 22(1): 63-67.
［6］He HC, Ling XH, Zhu JG, et al. Down-regulation of the ErbB3 binding protein 1 in human bladder cancer promotes tumor progression and cell proliferation［J］. Mol Biol Rep, 2013, 40(5): 3799-3805.
［7］Lessor TJ, Yoo JY, Xia X, et al. Ectopic expression of the ErbB-3 binding protein Ebp1 inhibits growth and induces differentiation of human breast cancer cell lines［J］. J Cell Physiol, 2000, 183(3): 321-329.
［8］Liu Z, Ahn J Y, Liu X, et al. Ebp1 isoforms distinctively regulate cell survival and differentiation［J］. Proce Natl Acad Sci USA, 2006, 103(29): 10917-10922.
［9］Friend C. Leukemia of adult mice caused by a transmissible agent［J］. Ann NY Acad Sci, 1957, 68(2): 522-532.
［10］Tsiftsoglou AS, Pappas IS, Vizirianakis IS. Mechanisms involved in the induced differentiation of leukemia cells［J］. Pharmacol Ther, 2003, 100(3): 257-290.
［11］Li L, Zhang G, Zhang Y, et al. Sodium butyrate-induced upregulation of p18(INK4C) gene affects k562 cell G(0)/G (1) arrest and differentiation［J］. Mol Cell Biochem, 2008, 319(1-2): 9-15.
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