Up-Regulation of Rho/ROCK Signaling in Sarcoma Cells Drives Invasion and Increased Generation of Protrusive Forces

Rösel, Daniel; Brábek, Jan; Tolde, Ondřej; Mierke, Claudia Tanja; Zitterbart, Daniel P.; Raupach, C.; Bicanová, Krisýtyna; Kollmannsberger, Philip; Paňková, Daniela; Veselý, Pavel; Folk, Petr; Fabry, Ben

doi:10.1158/1541-7786.mcr-07-2174

Cited by 98 publications

(76 citation statements)

References 37 publications

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“…In addition, the mesenchymal type of tumor cells (such as pII) have characteristics of fibroblast-like motility with elongated spindle-like shape and obvious leading edge towards the source of the chemotactic agent (72). On the other hand, an amoeboid-like movement was observed in various leukocytes (73,74) and tumor cells (75)(76)(77)(78) and is characterized by rounded shape of the cell during its motility. Cells with amoeboid type of movement have enhanced contractility and invasive capacity due to their rounded shape, which would conceivably enable them to squeeze through the small spaces in the extracellular matrix (ECM) fibers and exert higher force to deform the surrounding ECM (73)(74)(75)(76)(77).…”

Section: Discussionmentioning

confidence: 99%

Bleb formation is induced by alkaline but not acidic pH in estrogen receptor silenced breast cancer cells

Khajah

Mathew

Alam-Eldin

et al. 2015

International Journal of Oncology

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Abstract.De novo and acquired resistance to endocrinebased therapies in breast cancer occurs in parallel with epithelial to mesenchymal transition (EMT), which is associated with enhanced proliferative and metastatic potential, and poor clinical outcome. We have established several endocrine insensitive breast cancer lines by shRNA-induced depletion of estrogen receptor (ER) by transfection of MCF7 cells. All of these exhibit EMT. We have previously reported that brief exposure of specifically ER -breast cancer cells, to extracellular alkaline pH, results in cell rounding and segregation, and leads to enhanced invasive potential. In this study we describe more detailed morphological changes and compare these with cell exposure to acidic pH. Morphological changes and localization of various molecules critical for cell adhesion and motility, associated with pH effects, were assessed by live cell microscopy, electron microscopy, and immunofluorescence. Exposure of either ER -or ER + breast cancer cells to extracellular acidic pH did not induce significant changes in morphological appearance. Conversely, brief exposure of specifically ER silenced cells, to alkaline pH, resulted in cell contractolation and formation of bleb-like actin-rich structures which were evenly distributed on the outer membrane. Integrin α2, FAK, and JAM-1 were found in the cytoplasm streaming into the newly formed blebs. These blebs appear to be related to cell polarity and movement. Pre-treatment with cytochalasin-D or inhibitors of Rho or MLCK prevented both contractolation and bleb formation. Our data suggest that the effect of pH on the microenvironment of endocrine resistant breast cancer cells needs to be more extensively investigated. Alkaline, rather than acidic pH, appears to induce dramatic morphological changes, and enhances their invasive capabilities, through re-organization of cortical actin.

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Section: Discussionmentioning

confidence: 99%

Bleb formation is induced by alkaline but not acidic pH in estrogen receptor silenced breast cancer cells

Khajah

Mathew

Alam-Eldin

et al. 2015

International Journal of Oncology

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“…Overexpression of the ROCK proteins has been implicated in the invasive and metastatic potential of various tumor types [9][10][11]. Baseline protein expression of ROCK1 and ROCK2 was higher in H1975 cells compared to H1650 and H358 NSCLC cells (Fig.…”

Section: Role Of Rho-associated Coiled Coil Forming Protein (Rock) Anmentioning

confidence: 99%

Combined inhibition of rho-associated protein kinase and EGFR suppresses the invasive phenotype in EGFR-dependent lung cancer cells

et al. 2015

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a b s t r a c tIntroduction: Lung cancer remains the leading cause of cancer-related mortality worldwide, with metastatic disease frequently a prominent feature at the time of diagnosis. The role of NSCLC-derived EGFR mutations in cancer cell proliferation and survival has been widely reported, but little is known about the function of these mutations in invasive growth and metastasis. In this study, we sought to evaluate the intrinsic invasive properties of NSCLC cells with differing EGFR status and examine possible therapeutic targets that can abrogate invasive growth. Materials and methods: Collagen-based assays and 3D cell cultures were used to assess morphological features, actin cytoskeleton dynamics and the invasive capacity of NSCLC cell lines with differing EGFR status. The role of the RhoA/ROCK/MYPT1 and EGFR/HER pathways in NSCLC-related invasion was investigated by pharmacological inhibition and RNA interference techniques. Results: We demonstrate a positive correlation between EGFR mutational/amplification status and invasive capacity. Knockdown of wild-type and mutant EGFR leads to depletion of active and total MYPT1 levels. Combined pharmacological inhibition or genetic ablation of ROCK/EGFR suppresses the hallmarks of cancer cells and abrogates the invasive phenotype in EGFR-dependent NSCLC cells. Conclusions: These observations suggest that combined targeting of the ROCK and EGFR/HER pathways may be a potential therapeutic approach in limiting invasive growth in NSCLC.

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“…18 Additionally, ROCK/p-MLC is implicated in protrusive force generation at the leading edge of sarcoma cells. 24 ROCKs may drive retraction events to allow cells to reorient their direction of migration.…”

Section: A Possible Role For Rhoa In Integrin Clustersmentioning

confidence: 99%

Multiple roles for RhoA during T cell transendothelial migration

Heasman

Ridley

2010

Small GTPases

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t cells need to cross endothelial barriers during immune surveillance and inflammation. this involves t-cell adhesion to the endothelium followed by polarization and crawling with a lamellipodium at the front and contractile uropod at the back. t cells subsequently extend lamellipodia and filopodia under the endothelium in order to transmigrate. rho Gtpases play key roles in cell migration by regulating cytoskeletal dynamics and cell adhesion. we have found that the rho Gtpase rhoa is required for efficient t-cell polarization and migration on endothelial cells as well as transendothelial migration. rhoa-depleted cells lack both lamellipodia and uropods, and instead have narrow protrusions extending from a rounded cell body. using a rhoa activity biosensor, we have shown that rhoa is active at the leading edge in lamellipodia and filopodia of crawling and transmigrating t cells, as well as in the uropod. in lamellipodia, its activity correlates with both protrusion and retraction. we predict that rhoa signals via the formin mdia1 during lamellipodial protrusion whereas it induces lamellipodial retraction via the kinase rocK and actomyosin contractility. we propose that different guanine-nucleotide exchange factors (GEfs) are responsible for coordinating rhoa activation and signaling in different regions of transmigrating t cells. IntroductionDuring inflammation leukocytes are recruited from the vasculature into tissues using a specialized migratory pathway. Rho GTPase family proteins are key regulators of cytoskeletal dynamics and cell migration, 3 and thus would be expected to contribute to the process of TEM. Most Rho GTPases cycle between an inactive, GDP-bound form and an active, GTP-bound form, which interacts with and activates downstream targets. Rho GTPases are stimulated by guanine nucleotide exchange factors (GEFs), which stimulate exchange of GDP for GTP and inactivated by GTPaseactivating proteins (GAPs), which stimulate GTP hydrolysis.In our recent paper we used an RNAi screen to identify which Rho GTPases are important for T cell TEM. 4 We found that knockdown of RhoA induced the strongest inhibition of this process and so subsequently examined the role of RhoA in detail during both T cell crawling on EC and subsequent TEM. Significantly, active RhoA was observed at both the front and back of polarized T cells as they crawled on EC and transmigrated, as well as in dynamic puncta in the basal membrane and filopodia extending beneath the endothelium of transmigrating cells ( fig. 1). Here we discuss the implications of these findings in the context of current understanding of RhoA signaling and the potential function of these cellular processes in T-cell migration. Extra View to: Heasman SJ, Carlin LM, Cox S, Ng T, Ridley AJ. Coordinated RhoA signaling at the leading edge and uropod is required for T cell transendothelial migration.

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Up-Regulation of Rho/ROCK Signaling in Sarcoma Cells Drives Invasion and Increased Generation of Protrusive Forces

Cited by 98 publications

References 37 publications

Bleb formation is induced by alkaline but not acidic pH in estrogen receptor silenced breast cancer cells

Bleb formation is induced by alkaline but not acidic pH in estrogen receptor silenced breast cancer cells

Combined inhibition of rho-associated protein kinase and EGFR suppresses the invasive phenotype in EGFR-dependent lung cancer cells

Multiple roles for RhoA during T cell transendothelial migration

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