Selection of established tumour cells through narrow diameter micropores enriches for elevated Ras/Raf/MEK/ERK MAPK signalling and enhanced tumour growth.
Selection of established tumour cells through narrow diameter micropores enriches for elevated Ras/Raf/MEK/ERK MAPK signalling and enhanced tumour growth.
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DOI：
10.1080/21541248.2020.1780108
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发表时间：
2021-07
期刊：
影响因子：
--
通讯作者：
Olson MF
中科院分区：
文献类型：
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作者：
Rudzka DA;Mason S;Neilson M;McGarry L;Kalna G;Hedley A;Blyth K;Olson MF
随着正常细胞变成癌细胞，并向恶性发展，它们变得越来越软。这种变化的优点是肿瘤细胞变得更易变形，并且能够更好地通过狭窄的限制。我们设计了一种阳性选择策略，该策略富集可以通过窄直径微孔移动的细胞，以鉴定能够限制迁移的细胞表型。使用人MDA MB 231乳腺癌和MDA MB 435黑色素瘤癌细胞，我们发现，选择性选择有利于细胞具有相对较高的Ras/Raf/MEK/ERK丝裂原活化蛋白激酶（MAPK）信号，影响肌动蛋白细胞骨架组织，粘着斑密度和细胞弹性。在这项后续研究中，我们提供了进一步的证据表明，通过微孔选择富集的细胞具有改变的细胞形态和粘附。RNA测序数据的额外分析揭示了一组与小细胞大小相关的转录本，其独立于受约束的迁移。基因集富集分析确定了“基质体”作为最显着改变的基因集与小尺寸。当作为原位异种移植肿瘤在免疫功能低下的小鼠中生长时，经筛选的细胞比亲本或流式分选的细胞生长得显著更快。使用数学建模，我们确定了1）电池与间隙尺寸比; 2）电池的弯曲刚度之间存在相互作用，这使得能够通过窄间隙移动。这些结果扩展了我们先前的结论，Ras/Raf/MEK/ERK MAPK信号传导在调节细胞生物力学中具有重要作用，表明通过狭窄间隙的选择性运动压力也富集了体内肿瘤生长的增加。
As normal cells become cancer cells, and progress towards malignancy, they become progressively softer. Advantages of this change are that tumour cells become more deformable, and better able to move through narrow constraints. We designed a positive selection strategy that enriched for cells which could move through narrow diameter micropores to identify cell phenotypes that enabled constrained migration. Using human MDA MB 231 breast cancer and MDA MB 435 melanoma cancer cells, we found that micropore selection favoured cells with relatively higher Ras/Raf/MEK/ERK mitogen-activated protein kinase (MAPK) signalling, which affected actin cytoskeleton organization, focal adhesion density and cell elasticity. In this follow-up study, we provide further evidence that selection through micropores enriched for cells with altered cell morphology and adhesion. Additional analysis of RNA sequencing data revealed a set of transcripts associated with small cell size that was independent of constrained migration. Gene set enrichment analysis identified the ‘matrisome’ as the most significantly altered gene set linked with small size. When grown as orthotopic xenograft tumours in immunocompromised mice, micropore selected cells grew significantly faster than Parent or Flow-Sorted cells. Using mathematical modelling, we determined that there is an interaction between 1) the cell to gap size ratio; 2) the bending rigidity of the cell, which enable movement through narrow gaps. These results extend our previous conclusion that Ras/Raf/MEK/ERK MAPK signalling has a significant role in regulating cell biomechanics by showing that the selective pressure of movement through narrow gaps also enriches for increased tumour growth in vivo.