收稿日期: 2016-06-21
网络出版日期: 2018-06-28
基金资助
国家自然科学基金资助项目(31071283)
Effect of different Numb isoforms on proliferation and migration abilities of breast cancer cell line MCF-7
Received date: 2016-06-21
Online published: 2018-06-28
目的探讨Numb基因的4种不同亚型 对乳腺癌细胞增殖和迁移能力的影响.方法应用转录激活样效应因子核酸酶(transcription activator-like effector nuclease, TALEN)技术敲除乳腺癌细胞MCF-7中Numb基因, 分别将p65, p66, p71和p72这4种Numb亚型的表达载体转染进Numb-KO MCF-7细胞,应用CCK-8细胞增殖实验、细胞划痕实验、流式细胞术检测Numb基因及其4种 亚型对MCF-7胞增殖和迁移能力的影响.结果利用TALEN技术可以有效敲除MCF-7细胞中Numb基因,Numb-KO MCF-7组细胞的增殖和侵袭能力明显高于正常MCF-7细胞,Numb亚型p72对Numb-KO MCF-7细胞的增殖有明显的抑制作用,Numb亚型p65, p66, p71和p72均对Numb-KO MCF-7细胞的迁移有明显抑制作用.
苏杭, 王保垒, 李华顺, 朱晨光 . Numb基因不同亚型对乳腺癌细胞MCF-7增殖和迁移能力的影响[J]. 上海大学学报(自然科学版), 2018 , 24(3) : 476 -485 . DOI: 10.12066/j.issn.1007-2861.1822
Objective This study explores the role of Numb isoforms in proliferation and migration of breast cancer cells. Methods TALEN technology was used to knock out Numb gene in MCF-7 cells, and then transfected p65, p66, p71 and p72 expression vectors into these Numb-KO MCF-7 cells. On this occasion, CCK-8 proliferation assay and wound scratch assay were used to detect MCF-7 cell proliferation and migration. Results The results show that transcription activator-like effector nuclease (TALEN) is effective in knocking out Numb gene. Compared with normal MCF-7 cells, proliferation and migration abilities of Numb-KO MCF-7 cells are significantly enhanced. Numb isoform p72 inhibits proliferation of Numb-KO MCF-7 cells, and Numb isoforms p65, p66, p71 and p72 inhibit migration of Numb-KO MCF-7 cells.
Key words： breast cancer; Numb gene; isoform; cell proliferation; cell migration
| [1] | P ece S, C onfalonieri S, R r P, et al. Numb-ing down cancer by more than just a Notch[J]. Biochimica et Biophysica Acta, 2011,1815(1):26-43. |
| [2] | W ang Z, Li S S C. Numb: a new player in EMT[J]. Cell Adhesion and Migration, 2010,4(2):176-179. |
| [3] | D ing X, M a M, T eng J, et al. Numb induces E-cadherin adhesion dissolution, cytoskeleton reorganization, and migration in tubular epithelial cells contributing to renal fibrosis[J]. Current Molecular Medicine, 2015,15(4):368-379. |
| [4] | T osoni D, Z ecchini S, C oazzoli M, et al. The Numb/p53 circuitry couples replicative self-renewal and tumor suppression in mammary epithelial cells[J]. Journal of Cell Biology, 2015,211(4):845-862. |
| [5] | P ece S, S erresi M, S antolini E, et al. Loss of negative regulation by Numb over Notch is relevant to human breast carcinogenesis[J]. Journal of Cell Biology, 2004,167(2):215-221. |
| [6] | D i M L, F erretti E, G reco A, et al. Numb is a suppressor of Hedgehog signalling and targets Gli1 for itch-dependent ubiquitination[J]. Nature Cell Biology, 2006,8(12):1415-1423. |
| [7] | S hao X, L iu Y, Y u Q, et al. Numb regulates vesicular docking for homotypic fusion of early endosomes via membrane recruitment of Mon1b[J]. Cell Research, 2016,26(5):593-612. |
| [8] | S un M, A sghar S Z, Z hang H. The polarity protein Par3 regulates APP trafficking and processing through the endocytic adaptor protein Numb[J]. Neurobiology of disease, 2016,93(9):1-11. |
| [9] | D ho S E, F rench M B, W oods S A, et al. Characterization of four mammalian Numb protein isoforms. Identification of cytoplasmic and membrane-associated variants of the phosphotyrosine binding domain[J]. Journal of Biological Chemistry, 1999,274(46):33097-33104. |
| [10] | 钱文雨, 李华顺. Numb与神经发育[J]. 中国细胞生物学学报, 2010(1):1-16. |
| [11] | B ani-Y aghoub M, K ubu C J, C owling R, et al. A switch in Numb isoforms is a critical step in cortical development[J]. Developmental Dynamics, 2007,236(3):696-705. |
| [12] | V erdi J M, B ashirullah A, G oldhawk D E, et al. Distinct human Numb isoforms regulate differentiation vs. proliferation in the neuronal lineage[J]. Proceedings of the National Academy of Sciences of USA, 1999,96(18):10472-10476. |
| [13] | S leeman J P, T hiery J P. SNAPSHOT: the epithelial-mesenchymal transition[J]. Cell, 2011,145(1):162-162. |
| [14] | G ulino A, D i M L, S crepanti I. The multiple functions of Numb[J]. Experimental Cell Research, 2009,316(6):900-906. |
| [15] | K araczyn A, B aniyaghoub M, T remblay R, et al. Two novel human Numb isoforms provide a potential link between development and cancer[J]. Neural Development, 2010,5(12):1040-1048. |
| [16] | V erdi J M, B ashirullah A, G oldhawk D E. Distinct human Numb isoforms regulate differentiation vs. proliferation in the neuronal lineage[J]. Proceedings of the National Academy of Sciences, 1999,96(18):10472-10476. |
| [17] | T oriya M, T okunaga A, S awamoto K, et al. Distinct functions of human Numb isoforms revealed by misexpression in the neural stem cell lineage in the Drosophila larval brain[J]. Developmental Neuroscience, 2005,28(1/2):142-155. |
| [18] | G uo M, J an L Y, J an Y N. Control of daughter cell fates during asymmetric division: interaction of Numb and Notch[J]. Neuron, 1996,17(1):27-41. |
| [19] | D ho S E, J acob S, W olting C D, et al. The mammalian Numb phosphotyrosine-binding domain. Characterization of binding specificity and identification of a novel PDZ domain-containing Numb binding protein, LNX[J]. Journal of Biological Chemistry, 1998,273(15):9179-9187. |
| [20] | W illiamson M P. The structure and function of proline-rich regions in proteins[J]. Biochemical Journal, 1994,297(2):249-260. |
| [21] | B erdnik D, T orok T G M, K noblich J A. The endocytic protein alpha-adaptin is required for Numb-mediated asymmetric cell division in Drosophila[J]. Developmental Cell, 2002,3(2):221-231. |
| [22] | R asin M R, G azula V R, B reunig J J, et al. Numb and Numbl are required for maintenance of cadherin-based adhesion and polarity of neural progenitors[J]. Nature Neuroscience, 2007,10(7):819-827. |
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