Metalloprotease ADAMTS-1 decreases cell migration and invasion modulating the spatiotemporal dynamics of Cdc42 activity.
Metalloprotease ADAMTS-1 decreases cell migration and invasion modulating the spatiotemporal dynamics of Cdc42 activity.
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DOI：
10.1016/j.cellsig.2020.109827
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发表时间：
2021-01
影响因子：
4.8
通讯作者：
Freitas VM
中科院分区：
文献类型：
--
作者：
de Assis Lima M;da Silva SV;Serrano-Garrido O;Hülsemann M;Santos-Neres L;Rodríguez-Manzaneque JC;Hodgson L;Freitas VM
ADAMTS(具有血栓反应蛋白基序的去整合素和金属蛋白酶)是一种依赖于锌离子/钙离子的分泌型蛋白酶，参与生理和病理过程，是细胞外基质(ECM)的一部分。在这里，我们研究了ADAMTS-1是否是细胞侵袭和迁移所必需的，以及可能涉及的机制。为了检测ADAMTS-1‘S在卵巢癌细胞(CHO、NIH-OVCAR-3和ES2)和NIH-3T3成纤维细胞中的作用，我们对ADAMTS-1的表达水平进行了修饰，并与亲本进行了比较。与对照组相比，暴露于ADAMTS-1强化培养基中的细胞迁移和侵袭能力下降。当通过CRISPR/Cas9方法删除ADAMTS-1时，在细胞中观察到相反的情况。ADAMTS-1水平的下降增强了Src和FAK的磷酸化形式。我们还使用GLISA®试剂盒评估了细胞裂解物中细胞Rho GTP酶的活性。CDC42-GTP信号在CRISPR ADAMTS-1 ES-2细胞中显著增强。通过Förster共振能量转移(FRET)生物传感器检测ES-2细胞中CDC42的活性，我们发现与野生型细胞相比，ADAMTS-1缺失的迁移细胞前沿的CDC42活性被强烈极化。综上所述，ADAMTS-1抑制细胞增殖、极化和迁移。
ADAMTSs (A Disintegrin And Metalloproteinase with ThromboSpondin motifs) are secreted proteases dependent on Zn2+ / Ca2+, involved in physiological and pathological processes and are part of the extracellular matrix (ECM). Here, we investigated if ADAMTS-1 is required for invasion and migration of cells and the possible mechanism involved. In order to test ADAMTS-1’s role in ovarian cancer cells (CHO, NIH-OVCAR-3 and ES2) and NIH-3T3 fibroblasts, we modified the levels of ADAMTS-1 and compared those to parental. Cells exposed to ADAMTS-1-enriched medium exhibited a decline in cell migration and invasion when compared to controls with or without a functional metalloproteinase domain. The opposite was observed in cells when ADAMTS-1 was deleted via the CRISPR/Cas9 approach. The decline in ADAMTS-1 levels enhanced the phosphorylated form of Src and FAK. We also evaluated the activities of cellular Rho GTPases from cell lysates using the GLISA® kit. The Cdc42-GTP signal was significantly increased in the CRISPR ADAMTS-1 ES-2 cells. By a Förster resonance energy transfer (FRET) biosensor for Cdc42 activity in ES-2 cells we demonstrated that Cdc42 activity was strongly polarized at the leading edge of migrating cells with ADAMTS-1 deletion, compared to the wild type cells. As conclusion, ADAMTS-1 inhibits proliferation, polarization and migration.
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DOI：
10.1016/j.matbio.2018.06.002
发表时间：
2018-10
期刊：
Matrix biology : journal of the International Society for Matrix Biology
影响因子：
--
作者：
Mead TJ;Apte SS
通讯作者：
Apte SS
影响因子：
64.8
作者：
通讯作者：
--
DOI：
10.4049/jimmunol.1501655
发表时间：
2016-04-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Miskolci V;Wu B;Moshfegh Y;Cox D;Hodgson L
通讯作者：
Hodgson L
影响因子：
8
作者：
Liu, YJ;Xu, Y;Yu, Q
通讯作者：
Yu, Q
影响因子：
5
作者：
Lima, Maira Assis;dos Santos, Liliane;Freitas, Vanessa Morals
通讯作者：
Freitas, Vanessa Morals