Transition from mesenchymal to bleb‐based motility is predominantly exhibited by CD133‐positive subpopulation of fibrosarcoma cells

Abstract

Background Information Metastatic disease is caused by the ability of cancer cells to reach distant organs and form secondary lesions at new locations. Dissemination of cancer cells depends on their migration plasticity – an ability to switch between motility modes driven by distinct molecular machineries. One of such switches is mesenchymal‐to‐amoeboid transition. Although mesenchymal migration of individual cells requires Arp2/3‐dependent actin polymerisation, amoeboid migration is characterised by a high level of actomyosin contractility and often involves the formation of membrane blebs. The acquisition of amoeboid motility by mesenchymal cells is often associated with enhanced metastasis. Results We studied the ability of mesenchymal HT1080 fibrosarcoma cells to switch to amoeboid motility. We induced the transition from lamellipodium‐rich to blebbing phenotype either by down‐regulating the Arp2/3 complex, pharmacologically or by RNAi, or by decreasing substrate adhesiveness. Each of these treatments induced blebbing in a subset of fibrosarcoma cells, but not in normal subcutaneous fibroblasts. A significant fraction of HT1080 cells that switched to blebbing behaviour exhibited stem cell‐like features, such as expression of the stem cell marker CD133, an increased efflux of Hoechst‐33342 and positive staining for Oct4, Sox2 and Nanog. Furthermore, the isolated CD133+ cells demonstrated an increased ability to switch to bleb‐rich amoeboid phenotype both under inhibitor's treatment and in 3D collagen gels. Conclusions Together, our data show a significant correlation between the increased ability of cells to switch between migration modes and their stem‐like features, suggesting that migration plasticity is an additional property of stem‐like population of fibrosarcoma cells. This combination of features could facilitate both dissemination of these cells to distant locations, and their establishment self‐renewal in a new microenvironment, as required for metastasis formation. Significance These data suggest that migration plasticity is a new feature of cancer stem‐like cells that can significantly facilitate their dissemination to a secondary location by allowing them to adapt quickly to challenging microenvironments. Moreover, it complements their resistance to apoptosis and self‐renewal potential, thus enabling them not only to disseminate efficiently, but also to survive and colonise new niches.

Switching between blebbing and lamellipodia depends on the degree of non-muscle myosin II activity

“…We assessed myosin II activity in blebbing and lamellipodia cells by measuring pRLC/RLC ratio in the presence or absence of kinase inhibitors or genetic loss of these kinases. Genetic loss of ROCK and MLCK was carried out using RNA interference (RNAi) approach (Keller et al, 2012, Chikina et al, 2019a. As MDA-MB-231 cells express both ROCK1 and 2 isoforms, we used RNAi against both isoforms.…”

“…EsiRNA against MYLK (EHU158801), ROCK1 (EHU034701), and ROCK2 (EHU030131) and GFP (EHUEGFP) were procured from Sigma Aldrich. EsiRNA against GFP was used as a control for RNAi experiment (Keller et al, 2012, Chikina et al, 2019a). The siRNAs against integrinβ1, and Universal Negative Control 1 tagged with Cyanine 3 (NS siRNA, #SIC003) were purchased from Sigma Aldrich.…”

O-GlcNAcylation of ZEB1 facilitated mesenchymal pancreatic cancer cell ferroptosis

“…Accordingly, O-GlcNAcylation level significantly increased in the high glucose cultured PANC1 and PaTU8988 cells (Figure 1F, Figure S1E-F). Moreover, according to the previously publications, the mesenchymal human fibrosarcoma HT1080 cells [15], epithelial human breast cancer MCF7 cells [16] and human normal pancreatic epithelial cell line HPDE6 were chosen for the repeat experiments. The similar results can be acquired from mesenchymal HT1080 cells (Figure S1I-J), rather than epithelial MCF7 cells (Figure S1I-J) and normal HPDE6 cells (Figure S1G-H).…”

How does plasticity of migration help tumor cells to avoid treatment: Cytoskeletal regulators and potential markers

“…For more details on the main Arp2/3 inhibitors and their potential use as anti-tumor agents see review ( Chánez-Paredes et al, 2019 ). However, one must keep in mind that numerous studies show that suppression of actin polymerization in particular by inhibiting the action of the Arp2/3 complex stimulates the transition to amoeboid movement in cells of different types ( Beckham et al, 2014 ; Bergert et al, 2012 ; Chikina et al, 2019a , b ; Derivery et al, 2008 ; Logue et al, 2018 ; Obeidy et al, 2020 ). The authors of these studies suggest the weakening of the connection between the membrane and the submembrane actin cortex, or disruption of the regularity of this cortex, as the main triggers that stimulate MAP in cases of Arp2/3 suppression.…”