TRIM22靶向调控eIF4E在NB4细胞粒系定向分化过程中的作用与机制

doi:Leukemia; Cell differentiation; Eukaryotic initiation factor4E; Tripartite motif protein 22

国际肿瘤学杂志 ›› 2017, Vol. 44 ›› Issue (4): 251-256.doi: Leukemia; Cell differentiation; Eukaryotic initiation factor4E; Tripartite motif protein 22

TRIM22靶向调控eIF4E在NB4细胞粒系定向分化过程中的作用与机制

韩杨,宋冠华,田静,廖琼,李莲莲,张晓瑜,刘红艳,张之勇,姜国胜

250062 济南大学山东省医学科学院医学与生命科学学院（韩杨、田静、廖琼）；山东省医学科学院基础医学研究所（韩杨、宋冠华、田静、廖琼、李莲莲、张晓瑜、刘红艳、张之勇、姜国胜）

出版日期:2017-04-08 发布日期:2017-05-09
通讯作者: 姜国胜 E-mail:jiangguosh@163.com
基金资助:
国家自然科学基金(81573467)；山东省自然科学基金(ZR2015HM014)；山东省自主创新及成果转化专项(2014CGZH1313,2015ZDJS04003)；山东省泰山学者工程(ts201511075)；山东省科技攻关计划(2013YD18031)

Function and mechanism of TRIM22 targeting eIF4E in the process of NB4 cells differentiation

Han Yang, Song Guanhua, Tian Jing, Liao Qiong, Li Lianlian, Zhang Xiaoyu, Liu Hongyan, Zhang Zhiyong, Jiang Guosheng

School of Medicine and Life Sciences, University of Ji′nanShandong Academy of Medical Sciences; Institute of Basic Medicine, Shandong Academy of Medical Sciences, Ji′nan 250062, China

Online:2017-04-08 Published:2017-05-09
Contact: Jiang Guosheng E-mail:jiangguosh@163.com
Supported by:
National Natural Science Foundation of China (81573467); Natural Science Foundation of Shandong Province of China (ZR2015HM014); Shandong Province Key Project for Transformation of Results with Independent Innovation of China (2014CGZH1313, 2015ZDJS04003); Project for Laureate of Taishan Scholar (ts201511075); Key Science and Technology Program of Shandong Province of China (2013YD18031)

摘要/Abstract

摘要： 目的探讨在人急性早幼粒细胞白血病NB4细胞粒系分化过程中三基序蛋白22（TRIM22）的功能及其与真核细胞起始因子4E（eIF4E）的相互作用，从而进一步阐明TRIM22靶向调控eIF4E的机制。方法体外实验建立NB4细胞诱导分化模型，应用实时荧光定量聚合酶链反应(qRTPCR)及Western blotting技术检测TRIM22、eIF4E基因和蛋白表达的变化，并通过电穿孔转染技术敲低或过表达TRIM22后利用CCK8和流式细胞术检测其对细胞功能的影响，及对eIF4E蛋白表达水平的影响，最后利用免疫共沉淀（COIP）实验验证TRIM22与eIF4E的相互作用。结果全反式维甲酸（ATRA）诱导NB4细胞后，TRIM22的mRNA相对表达量由1.01±0.15逐渐升高到30.98±2.79（F=280.700，P=0.000），蛋白相对表达水平由0.22±0.03逐渐升高至0.51±0.05（F=51.430，P=0.000）。反之，eIF4E的mRNA相对表达量由1.01±0.09逐渐降低至0.47±0.06（F=20.520，P=0.000），蛋白相对表达水平由0.97±0.02逐渐降低至0.64±0.09（F=14.700，P=0.001）。流式细胞术检测TRIM22过表达后ATRA干预组PECD11b表达水平［(78.80±2.00)%］比空载体ATRA干预组［(58.70±2.70)%］和共转染后ATRA干预组［(61.60±3.80)%］均升高（t=9.535，P=0.000；t=8.187，P=0.000）。同时，过表达TRIM22基因水平后，eIF4E蛋白水平会反向变化(t=4.985，P=0.007)。COIP实验验证TRIM22通过结合eIF4E而起作用。结论 TRIM22在NB4细胞粒系定向分化过程中促进细胞分化，并且可通过靶向结合eIF4E抑制其表达而发挥重要作用。

关键词: 白血病, 细胞分化, 真核细胞起始因子4E, 三基序蛋白22

Abstract: Objective To investigate the function of tripartite motif protein 22 (TRIM22) and the interaction with eukaryotic translation initiation factor4E (eIF4E) in the differentiation of NB4 cells, one kind of acute promyelocytic leukemia cells, which elucidates the mechanism of TRIM22 targeting to regulate eIF4E. Methods The model of NB4 cells inducing differentiation was established in vitro. The expression changes of gene and protein of TRIM22 and eIF4E were detected by using quantitative realtime polymerase chain reaction (qRTPCR) and Western blotting. In addition, the effect on cell function and protein expression level of eIF4E after adopting electroporation technology to depress or overexpress TRIM22 was detected by CCK8 and flow cytometry. Finally, the interaction of TRIM22 and eIF4E was verified by using coimmunoprecipitation (COIP). Results The mRNA relative expression level of TRIM22 was gradually increasing from 1.01±0.15 to 30.98±2.79 (F=280.700, P=0.000), and the protein relative expression level was gradually increasing from 0.22±0.03 to 0.51±0.05 (F=51.430, P=0.000) after the alltransretinoic acid (ATRA) induction for NB4 cell. However, the mRNA relative expression level of eIF4E was gradually decreasing from 1.01±0.09 to 0.47±0.06 (F=20.520, P=0.000), with the same trend, the protein relative expression level was gradually decreasing from 0.97±0.02 to 0.64±0.09 (F=14.700, P=0.001). The expression level of PECD11b in the TRIM22 overexpression group with ATRA detected by flow cytometry ［(78.80±2.00)%］ was higher than that in the transfection group of empty vetor with ATRA ［(58.70±2.70)%］ (t=9.535, P=0.000) and the cotransfection group with ATRA ［(61.60±3.80)%］ (t=8.187, P=0.000). Meanwhile, the protein level of eIF4E changed reversely after overexpressing the gene level of TRIM22 (t=4.985, P=0.007). The COIP experiment was used to verify the interaction of TRIM22 and eIF4E. Conclusion TRIM22 is able to promote the cell differentiation during the process of NB4 cells differentiation. Furthermore, eIF4E is a target of TRIM22 for binding with, which plays an important role in depressing the expression of eIF4E

Key words: , Leukemia, Cell differentiation, Eukaryotic initiation factor-4E, Tripartite motif protein 22

韩杨,宋冠华,田静,廖琼,李莲莲,张晓瑜,刘红艳,张之勇,姜国胜. TRIM22靶向调控eIF4E在NB4细胞粒系定向分化过程中的作用与机制[J]. 国际肿瘤学杂志, 2017, 44(4): 251-256.

Han Yang, Song Guanhua, Tian Jing, Liao Qiong, Li Lianlian, Zhang Xiaoyu, Liu Hongyan, Zhang Zhiyong, Jiang Guosheng. Function and mechanism of TRIM22 targeting eIF4E in the process of NB4 cells differentiation[J]. Journal of International Oncology, 2017, 44(4): 251-256.

参考文献

［1］ Hattlmann CJ, Kelly JN, Barr SD. TRIM22: a diverse and dynamic antiviral protein［J］. Mol Biol Int, 2012, 2012: 153415. DOI: 10.1155/2012/153415. ［2］ Yap MW, Stoye JP. TRIM proteins and the innate immune response to viruses［J］. Adv Exp Med Biol, 2012, 770: 93104. ［3］ Sun Y, Ho GH, Koong HN, et al. Downregulation of Tripartitemotif containing 22 expression in breast cancer is associated with a lack of p53mediated induction［J］. Biochem Biophys Res Commun, 2013, 441(3): 600606. DOI: 10.1016/j.bbrc.2013.10.110. ［4］ Singh R, Gaiha G, Werner L, et al. Association of TRIM22 with the type 1 interferon response and viral control during primary HIV1 infection［J］. J Virol, 2011, 85(1): 208216. DOI: 10.1128/JVI.0181010. ［5］ Ovsyannikova IG, Haralambieva IH, Vierkant RA, et al. Associations between polymorphisms in the antiviral TRIM genes and measles vaccine immunity［J］. Hum Immunol, 2013, 74(6): 768774. DOI: 10.1016/j.humimm.2013.01.031. ［6］ Topisirovic I, Svitkin YV, Sonenberg N, et al. Cap and capbinding proteins in the control of gene expression［J］. Wiley Interdiscip Rev RNA, 2011, 2(2): 277298. DOI: 10.1002/wrna.52. ［7］ Zong AZ, Liu YH, Zhang Y, et al. Antitumor activity and the mechanism of SIPS: a sulfated polysaccharide with antimetastatic effect［J］. Carbohydr Polym, 2015, 129: 5054. DOI: 10.1016/j.carbpol.2015.04.017. ［8］ Khosravi S, Tam KJ, Ardekani GS, et al. eIF4E is an adverse prognostic marker of melanoma patient survival by increasing melanoma cell invasion［J］. J Invest Dermatol, 2015, 135(5): 13581367. DOI: 10.1038/jid.2014.552. ［9］ Volpon L, Osborne MJ, Zahreddine H, et al. Conformational changes induced in the eukaryotic translation initiation factor eIF4E by a clinically relevant inhibitor, ribavirin triphosphate［J］. Biochem Biophys Res Commun, 2013, 434(3): 614619. DOI: 10.1016/j.bbrc.2013.03.125. ［10］ Carroll M, Borden KL. The oncogene eIF4E: using biochemical insights to target cancer［J］. J Interferon Cytokine Res, 2013, 33(5): 227238. DOI: 10.1089/jir.2012.0142. ［11］ Willimott S, Beck D, Ahearne MJ, et al. Captranslation inhibitor, 4EGI1, restores sensitivity to ABT737 apoptosis through capdependent and independent mechanisms in chronic lymphocytic leukemia［J］. Clin Cancer Res, 2013, 19(12): 32123223. DOI: 10.1158/10780432.CCR122185. ［12］ Kentsis A, Dwyer EC, Perez JM, et al. The RING domains of the promyelocytic leukemia protein PML and the arenaviral protein Z repress translation by directly inhibiting translation initiation factor eIF4E［J］. J Mol Biol, 2001, 312(4): 609623. DOI: 10.1006/jmbi.2001.5003. ［13］ Rajsbaum R, GarciaSastre A, Versteeg GA. TRIMmunity: the roles of the TRIM E3Ubiquitin ligase family in innate antiviral immunity［J］. J Mol Biol, 2014, 426(6): 12651284. DOI: 10.1016/j.jmb.2013.12.005. ［14］ Ndung′u T. TRIM E3 ligases in HIV infection: can these intrinsic immunity factors be harnessed for novel antiviral strategies?［J］. Virulence, 2011, 2(4): 360366. ［15］ KajasteRudnitski A, Marelli SS, Pultrone C, et al. TRIM22 inhibits HIV1 transcription independently of its E3 ubiquitin ligase activity, Tat, and NFkappaBresponsive long terminal repeat elements［J］. J Virol, 2011, 85(10): 51835196. DOI: 10.1128/JVI.0230210.

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TRIM22靶向调控eIF4E在NB4细胞粒系定向分化过程中的作用与机制

Function and mechanism of TRIM22 targeting eIF4E in the process of NB4 cells differentiation

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