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NOTCH1对T-ALL抑癌基因ID4的负调控作用

刘娟, 燕莞瑶, 楼一层

刘娟, 燕莞瑶, 楼一层. NOTCH1对T-ALL抑癌基因ID4的负调控作用[J]. 肿瘤防治研究, 2016, 43(2): 123-128. DOI: 10.3971/j.issn.1000-8578.2016.02.006
引用本文: 刘娟, 燕莞瑶, 楼一层. NOTCH1对T-ALL抑癌基因ID4的负调控作用[J]. 肿瘤防治研究, 2016, 43(2): 123-128. DOI: 10.3971/j.issn.1000-8578.2016.02.006
LIU Juan, YAN Wanyao, LOU Yiceng. NOTCH1 Down-regulates Tumor Suppressor Gene ID4 in T-cell Acute Lymphoblastic Leukemia[J]. Cancer Research on Prevention and Treatment, 2016, 43(2): 123-128. DOI: 10.3971/j.issn.1000-8578.2016.02.006
Citation: LIU Juan, YAN Wanyao, LOU Yiceng. NOTCH1 Down-regulates Tumor Suppressor Gene ID4 in T-cell Acute Lymphoblastic Leukemia[J]. Cancer Research on Prevention and Treatment, 2016, 43(2): 123-128. DOI: 10.3971/j.issn.1000-8578.2016.02.006

NOTCH1对T-ALL抑癌基因ID4的负调控作用

基金项目: 国家自然科学基金(30870249,81173509)
详细信息
    作者简介:

    刘娟(1989-),女,硕士在读,主要从事急性T淋巴细胞白血病的致病机制研究

    通讯作者:

    楼一层,E-mail:louyiceng@163.com

  • 中图分类号: R392.12; R733.71

NOTCH1 Down-regulates Tumor Suppressor Gene ID4 in T-cell Acute Lymphoblastic Leukemia

  • 摘要: 目的 研究DNA结合抑制因子4(inhibitor of DNA binding 4, ID4)在急性T淋巴细胞白血病(T-cell acute lymphoblastic leukemia, T-ALL)中的表达水平,探讨NOTCH1与ID4的相关性及其调控机制。方法 通过Oncomine和Pieters R数据库分析T-ALL患者与正常捐献者中ID4的表达差异及T-ALL患者中NOTCH1与ID4的相关性,用qRT-PCR和Western blot检测阻断NOTCH1或激活NOTCH1后ID4的表达变化进行验证。用生物信息学的方法特异预测出microRNA-342(miR-342)靶向ID4,通过双荧光素酶报告体系进行验证。过表达或沉默miR-342后,qRT-PCR和Western blot检测ID4的表达变化。阻断NOTCH1或激活NOTCH1,qRT-PCR检测miR-342的表达变化。结果 ID4在T-ALL患者中的表达较正常组显著下调(P<0.01);ID4受NOTCH1负调控(P<0.05);双荧光素酶报告系统验证预测结果正确;过表达miR-342后,ID4的表达受到显著抑制(P<0.05),沉默miR-342后,ID4的表达明显上调(P<0.05);NOTCH1信号的阻断能够抑制miR-342的表达,NOTCH1信号的激活能够上调miR-342的表达。结论 NOTCH1通过激活miR-342对ID4的负调控作用从而下调抑癌基因ID4的表达,促进T-ALL的发生。

     

    Abstract: Objective To explore the expression level of inhibitor of DNA binding 4(ID4) in T-cell acute lymphoblastic leukemia(T-ALL), and to determine the correlation and mechanism between NOTCH1 and ID4. Methods The expression levels of ID4 in T-ALL patients and normal donors, as well as the correlation of NOTCH1 and ID4, were analyzed from the databases of Oncomine and Pieters R. ID4 mRNA and protein levels were detected by qRT-PCR and Western blot after NOTCH1 inhibition or activation, which were adopted to verify the correlation. MicroRNA-342 (miR-342) was predicted to target ID4 with the approach of bioinformatics, and the dual luciferase reporter system was carried out to verify the prediction. ID4 mRNA and protein levels were detected by qRT-PCR after overexpressing or silencing miR-342. MiR-342 expression level was detected by qRT-PCR and Western blot after NOTCH1 inhibition or activation. Results ID4 expression level in T-ALL patients was significantly lower than that in normal donors(P<0.01); ID4 was negatively regulated by NOTCH1(P<0.05); The dual luciferase reporter system validated ID4 as a specific target gene of miR-342; Down-regulation of miR-342 resulted in sharp increase of ID4 mRNA and protein (P<0.05), and upregulation of miR-342 resulted in sharp decrease of ID4 mRNA and protein (P<0.05); NOTCH1 inhibition could down-regulate the expression of miR-342, and NOTCH1 activation could up-regulate the expression of miR-342. Conclusion NOTCH1 activates the negatively regulatory role of miR-342 for ID4, then downregulates tumor suppressor gene ID4 expression, and may contribute to the pathogenesis of T-ALL.

     

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出版历程
  • 收稿日期:  2015-09-07
  • 修回日期:  2015-09-07
  • 刊出日期:  2016-02-24

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