Numb在三阴乳腺癌中抑制HDM2泛素化降解p53 <sup>*</sup>

doi:10.13523/j.cb.20190701

中国生物工程杂志

2019, Vol. 39

Issue (7): 1-7 DOI: 10.13523/j.cb.20190701

研究报告

Numb在三阴乳腺癌中抑制HDM2泛素化降解p53 ^*

先洁,覃雪,曹友德(

)

重庆医科大学基础医学院重庆 400016

Numb Inhibits the Ubiquitination Degradation of p53 by HDM2 in Triple-negative Breast Cancer

Jie XIAN,Xue QIN,You-de CAO(

)

Teaching and Research Section of Pathology, School of Basic Medical Sciences, Medical University of Chongqing, Chongqing 400016, China

全文: PDF(941 KB) HTML

摘要：

目的探究Numb蛋白在三阴乳腺癌患者中的表达降低情况,及Numb蛋白在三阴乳腺癌中对抑癌因子p53蛋白水平的影响及调控机制,进一步研究Numb蛋白的降低与三阴乳腺癌发生发展的相关性,从而为缺乏有效治疗方法的三阴乳腺癌提供一个潜在的治疗新靶点。方法 40例三阴乳腺癌患者病理组织切片取自重庆医科大学临床病理诊断中心,采用免疫组化法检测Numb蛋白在三阴乳腺癌患者中的表达情况。MCF-10A细胞株和MDA-MB-231细胞株均为ATCC来源,采用qPCR和Western blot法检测对比Numb、HDM2、p53三者的转录水平和蛋白质水平在以上两个细胞株中差异。采用增强型绿色荧光蛋白(enhance green fluorescent protein, EGFP)质粒转染的方法在MDA-MB-231细胞中重表达Numb,采用qPCR和Western blot法验证Numb、HDM2、p53三者表达的变化。结果转染NUMB-EGFP后MDA-MB-231细胞中Numb的mRNA和蛋白质水平均明显上调,HDM2无显著改变,p53在转录水平无明显变化,但在蛋白质水平显著升高。在231细胞中上调Numb蛋白可以在转录后水平调节p53水平,使p53蛋白随之显著升高。结论 Numb蛋白在三阴乳腺癌患者中表达降低的比列很高,为55%,且Numb蛋白在三阴乳腺癌细胞MDA-MB-231中可以调控抑癌因子p53蛋白水平,Numb蛋白水平与p53蛋白水平呈正相关。

关键词： 三阴乳腺癌; Numb蛋白; HDM2蛋白; p53蛋白

Abstract:

Objective : To investigate the decrease of Numb protein expression in triple-negative breast cancer patients and the effect of Numb protein on the p53 protein levels in triple-negative breast cancer and its regulation mechanism, and to further study the correlation between the decrease of Numb protein and the occurrence and development of triple-negative breast cancer, so as to provide a potential new therapeutic target for triple-negative breast cancer without effective treatment.Methods: 40 cases of triple negative breast cancer were taken from Clinical pathology diagnosis centre of Medical University of Chongqing. The expression of Numb protein in triple negative breast cancer patients was detected by immunohistochemistry. MCF-10A cell line and MDA-MB-231 cell line were all ATCC-derived. The mRNA and protein levels of Numb, HDM2 and p53 were compared by qPCR and Western blot. Numb was re-expressed in MDA-MB-231 cells by plasmid transfection. The expression of Numb, HDM2 and p53 was verified by qPCR and Western blot.Results: After transfection with NUMB-EGFP, the mRNA and protein levels of Numb in MDA-MB-231 cells were significantly increased, HDM2 did not change evidently, and p53 did not change significantly at the mRNA levels, but increased remarkably at the protein levels. The increased of Numb protein in NUMB-EGFP-transfected cells can regulate the p53 levels at post-transcriptional level, resulting in a significant increase of p53 protein levels.Conclusion: The expression of Numb protein in triple-negative breast cancer patients decreased by 55%, and Numb protein can regulate the level of p53 protein in triple-negative breast cancer cell line MDA-MB-231. The level of Numb protein is positively correlated with the level of p53 protein.

Key words: Triple-negative breast cancer Numb protein HDM2 protein p53 protein

收稿日期: 2019-01-16 出版日期: 2019-08-05

ZTFLH:

Q819

基金资助: * 重庆市科委重庆市社会事业与民生保障科技创新专项资助项目(cstc2016shms-ztzx130001)

通讯作者: 曹友德 E-mail: 420306100@qq.com

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	先洁
	覃雪
	曹友德

引用本文:

先洁,覃雪,曹友德. Numb在三阴乳腺癌中抑制HDM2泛素化降解p53 ^*[J]. 中国生物工程杂志, 2019, 39(7): 1-7.

Jie XIAN,Xue QIN,You-de CAO. Numb Inhibits the Ubiquitination Degradation of p53 by HDM2 in Triple-negative Breast Cancer. China Biotechnology, 2019, 39(7): 1-7.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20190701 或 https://manu60.magtech.com.cn/biotech/CN/Y2019/V39/I7/1

表1 引物序列

图1 Numb蛋白在三阴乳腺癌患者组中表达不均,明显低于人正常乳腺组织组

表2 Numb蛋白的表达情况与三阴乳腺癌患者的临床特征之间的关联

图2 Numb、HDM2、p53三者转录和蛋白质水平在MCF-10A和MDA-MB-231细胞株中的差异

图3 在MDA-MB-231细胞株中过表达Numb蛋白

[1]	Pece S, Serresi M, Santolini E , et al. Loss of negative regulation by Numb over notch is relevant to human breast carcinogenesis. J Cell Biol, 2004,167(2):215-221. doi: 10.1083/jcb.200406140
[2]	Di Marcotullio L, Ferretti E, Greco A , et al. Numb is a suppressor of hedgehog signallingand targets gli1 for itch-dependent ubiquitination. Nature , 2006,8(12):1415-1423.
[3]	Cahilly-Snyder L, Yang-Feng T, Francke U , et al. Molecular analysis and chromosomal mapping of amplified genes isolated from a transformed mouse 3T3 cell line. Somat Cell Mol Genet, 1987,13(3):235-244. doi: 10.1007/BF01535205
[4]	Vousden K H, Prives C . P53 and prognosis:new insights and further complexity. Cell, 2005,120(1):7-10.
[5]	Momand J, Wu H H, Dasgupta G , et al. MDM2-master regulator of the p53 tumor suppressor protein. Gene, 2000,242(1/2):15-29. doi: 10.1016/S0378-1119(99)00487-4
[6]	Hutterer A, Knoblich J A . Numb and a-adaptin regulate sanpodo endocytosis to specify cell fate in drosophilaexternal sensoryorgans. EMBO Rep, 2005,6(9):836-842. doi: 10.1038/sj.embor.7400500
[7]	Foulkes W D, Smith I E, Reis-Filho J S , et al. Triple-negative breast cancer. N Engl J Med, 2010,363(20):1938-1948. doi: 10.1056/NEJMra1001389
[8]	Pece S, Confalonieri S, Di Fiore P P , et al. NUMB-ing down cancer by more than just a NOTCH. Biochim Biophys Acta, 2011,1815(1):26-43.
[9]	Shen Q, Zhong W, Jan Y , et al. Asymmetric numb distribution is critical for asymmetric cell division of mouse cerebral cortical stem cells and neuroblasts. Development , 2002,129(20):4843-4853.
[10]	Colaluca I N, Tosoni D, Nuciforo P , et al. NUMB controls p53 tumour suppressor activity. Nature, 2008,451(7174):76-81.
[11]	Zhang Y, Li F Y, Song Y L , et al. Numb and Numbl act to determine mammary myoepithelial cell fate,maintain epithelial identity,and support lactogenesis. FASEB, 2016,30(10):3474-3488. doi: 10.1096/fj.201600387R
[12]	Rhyu M S, Jan L Y, Jan Y N . Asymmetric distribution of numb protein during division of the sensory organ precursor cell confers distinct fates to daughter cells. Cell, 1994,76(3):477-491. doi: 10.1016/0092-8674(94)90112-0
[13]	Juven-Gershon T, Shifman O, Unger T , et al. The mdm2 oncoprotein interacts with the cell fate regulator Numb. Mol Cell Biol, 1998,18(7):3974-3982. doi: 10.1128/MCB.18.7.3974
[14]	Berdnik D, Török T, Gonzalez-Gaitan M , et al. The endocytic proteina-adaptin is required for numb-mediated asymmetric cell division in drosophila. Dev Cell , 2002,3(2):221-231. doi: 10.1016/S1534-5807(02)00215-0
[15]	Vassilev L T, Vu B T, Graves B , et al. In vivo activation of the p53 pathway by small-molecule antagonists of MDM2. Science, 2004,303(5659):844-848. doi: 10.1126/science.1092472
[16]	Tosoni1 D, Pambianco1 S, Ekalle Soppo B , et al. Re-clinical validation of a selective anti-cancer stem cell therapy for Numb-deficient human breast cancer. EMBO Mol Med, 2017,7(5):656-671.
[17]	Ray-Coquard I, Blay J Y, Italiano A , et al. Effect of the MDM2 antagonist RG7112 on the P53 pathway in patients with MDM2-amplified, well-differentiated or dedifferentiated liposarcoma: an exploratory proof-of-mechanism study. Lancet Oncol , 2012,13(11):1133-1140. doi: 10.1016/S1470-2045(12)70474-6
[18]	Pece S, Confalonieri S, R Romano P , et al. NUMB-ing downcancer by more than just a NOTCH. Biochim Biophys Acta, 2011,1815(1):26-43.
[19]	Enari M, Ohmori K, Kitabayashi I , et al. Requirement of clathrin heavy chain for p53-mediated transcription. Genes Dev, 2006,20(9):1087-1099. doi: 10.1101/gad.1381906
[20]	Cayouette M, Raff M , et al. Asymmetric segregation of Numb: a mechanism for neural specification from Drosophila to mammals. Nature Neurosci, 2002,5(12):1265-1269.

[1]	林敏. 玉米生物育种基础研究与关键技术[J]. 中国生物工程杂志, 2021, 41(12): 1-3.
[2]	吴函蓉,王莹,黄英明,李冬雪,李治非,方子寒,范玲. 以基地平台为抓手,促进生物技术创新与转化[J]. 中国生物工程杂志, 2021, 41(12): 141-147.
[3]	尹泽超,王晓芳,龙艳,董振营,万向元. 玉米穗腐病抗性鉴定、遗传分析与分子机制^*[J]. 中国生物工程杂志, 2021, 41(12): 103-115.
[4]	冷燕,孙康泰,刘倩倩,蒲阿庆,李翔,万向元,魏珣. 全球基因编辑作物监管趋势研究[J]. 中国生物工程杂志, 2021, 41(12): 24-29.
[5]	何伟,祝蕾,刘欣泽,安学丽,万向元. 玉米遗传转化与商业化转基因玉米开发^*[J]. 中国生物工程杂志, 2021, 41(12): 13-23.
[6]	杨梦冰,江易林,祝蕾,安学丽,万向元. CRISPR/Cas植物基因组编辑技术及其在玉米中的应用^*[J]. 中国生物工程杂志, 2021, 41(12): 4-12.
[7]	殷芳冰,王成,龙艳,董振营,万向元. 玉米雌穗性状遗传分析与形成机制^*[J]. 中国生物工程杂志, 2021, 41(12): 30-46.
[8]	秦文萱,刘鑫,龙艳,董振营,万向元. 玉米叶夹角形成的遗传基础与分子机制解析^*[J]. 中国生物工程杂志, 2021, 41(12): 74-87.
[9]	王锐璞,董振营,高悦欣,龙艳,万向元. 玉米籽粒淀粉含量遗传基础与调控机制^*[J]. 中国生物工程杂志, 2021, 41(12): 47-60.
[10]	马雅杰,高悦欣,李依萍,龙艳,董振营,万向元. 玉米株高和穗位高的遗传基础与分子机制^*[J]. 中国生物工程杂志, 2021, 41(12): 61-73.
[11]	王彦博,魏佳,龙艳,董振营,万向元. 玉米雄穗性状遗传结构与形成分子机制^*[J]. 中国生物工程杂志, 2021, 41(12): 88-102.
[12]	毛开云,李荣,李丹丹,赵若春,范月蕾,江洪波. 全球双特异性抗体药物研发格局分析^*[J]. 中国生物工程杂志, 2021, 41(11): 110-118.
[13]	吴函蓉,王莹,杨力,葛瑶,范玲. 我国生物技术基地平台现状与发展建议[J]. 中国生物工程杂志, 2021, 41(11): 119-123.
[14]	刘天义,冯卉,SALSABEELYousuf,解领丽,苗向阳. lncRNA在动物脂肪沉积中的研究进展^*[J]. 中国生物工程杂志, 2021, 41(11): 82-88.
[15]	薛志勇,代红生,张显元,孙艳颖,黄志伟. 表达透明颤菌血红蛋白基因对酿酒酵母生长及细胞内氧化状态的影响^*[J]. 中国生物工程杂志, 2021, 41(11): 32-39.

Viewed

Full text

Abstract

Cited

Shared

Discussed