The effects of modifying RhoA and Rac1 activities on heterotypic contact inhibition of locomotion

Anear, Erika; Parish, Roger W.

doi:10.1016/j.febslet.2012.03.044

Cited by 14 publications

(14 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…When fibroblasts with normal motility collide, they undergo temporary paralysis of movement and then divert their paths to avoid contact in a phenomenon known as contact inhibition of locomotion, which was first described by Michael Abercrombie and Joan Heaysman in the early 1950s [90,91]. The cell overlap shown by IRM here indicates that ROCK inhibition causes a release from contact inhibition of locomotion, as reported before [92][93][94], and reviewed by Mayor and Carmona-Fontaine [95]. A consequence of contact inhibition of locomotion is that sparse cells will translocate faster than more numerous cells due to a lesser number of collisions [90].…”

Section: Functional Consequences Of Irs-1 Inhibitionsupporting

confidence: 66%

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

Czarnowski

Papp

Száraz

et al. 2014

Cellular and Molecular Biology Letters

View full text Add to dashboard Cite

Cellular adhesion to the underlying substratum is regulated through numerous signaling pathways. It has been suggested that insulin receptor substrate 1 (IRS-1) is involved in some of these pathways, via association with and activation of transmembrane integrins. Calreticulin, as an important endoplasmic reticulum-resident, calcium-binding protein with a chaperone function, plays an obvious role in proteomic expression. Our previous work showed that calreticulin mediates cell adhesion not only by affecting protein expression but also by affecting the state of regulatory protein phosphorylation, such as that of c-src. Here, we demonstrate that calreticulin affects the abundance of IRS-1 such that the absence of calreticulin is paralleled by a decrease in IRS-1 levels and the unregulated overexpression of calreticulin is accompanied by an increase in IRS-1 levels. These changes in the abundance of calreticulin and IRS-1 are accompanied by changes in cell-substratum adhesiveness and phosphorylation, such that increases in the expression of calreticulin and IRS-1 are paralleled by an increase in focal contact-based cellsubstratum adhesiveness, and a decrease in the expression of these proteins brings about a decrease in cell-substratum adhesiveness. Wild type and calreticulin-null mouse embryonic fibroblasts (MEFs) were cultured and the IRS-1 isoform profile was assessed. Differences in morphology and motility were also quantified. While no substantial differences in the speed of locomotion were found, the directionality of cell movement was greatly promoted by the presence of calreticulin. Calreticulin expression was also found to have a dramatic effect on the phosphorylation state of serine 636 of IRS-1, such that phosphorylation of IRS-1 on serine 636 increased radically in the absence of calreticulin. Most importantly, treatment of cells with the RhoA/ROCK inhibitor, Y-27632, which among its many effects also inhibited serine 636 phosphorylation of IRS-1, had profound effects on cell-substratum adhesion, in that it suppressed focal contacts, induced extensive close contacts, and increased the strength of adhesion. The latter effect, while counterintuitive, can be explained by the close contacts comprising labile bonds but in large numbers. In addition, the lability of bonds in close contacts would permit fast locomotion. An interesting and novel finding is that Y-27632 treatment of MEFs releases them from contact inhibition of locomotion, as evidenced by the invasion of a cell's underside by the thin lamellae and filopodia of a cell in close apposition.

show abstract

Section: Functional Consequences Of Irs-1 Inhibitionsupporting

confidence: 66%

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

Czarnowski

Papp

Száraz

et al. 2014

Cellular and Molecular Biology Letters

View full text Add to dashboard Cite

show abstract

“…EphA receptor activation has been shown to lead to cell retraction via RhoA in several cell types (Astin et al, 2010; Lawrenson et al, 2002; Ogita et al, 2003; Shamah et al, 2001; Wahl et al, 2000) and RhoA has been implicated in CIL (Anear and Parish, 2012; Carmona-Fontaine et al, 2008; Kadir et al, 2011; Theveneau et al, 2010). However, the functional requirement for RhoA in prostate cancer contact repulsion has not been shown.…”

Section: Resultsmentioning

confidence: 99%

“…Several studies have shown that Rho GTPases play key roles during CIL (Anear and Parish, 2012; Astin et al, 2010; Carmona-Fontaine et al, 2008; Kadir et al, 2011; Theveneau et al, 2010). Rho GTPases are key regulators of the actin and microtubule cytoskeleton and cell polarity, and may regulate all of the key repulsion steps during CIL.…”

Section: Discussionmentioning

confidence: 99%

EphA receptors regulate prostate cancer cell dissemination through Vav2–RhoA mediated cell–cell repulsion

et al. 2014

View full text Add to dashboard Cite

Metastatic prostate cancer cells display EphB receptor-mediated attraction when they contact stromal fibroblasts but EphA-driven repulsion when they contact one another. The impact of these ‘social’ interactions between cells during cancer cell invasion and the signalling mechanisms downstream of Eph receptors are unclear. Here we show that EphA receptors regulate prostate cancer cell dissemination in a 2D dispersal assay and in a 3D cancer cell spheroid assay. We show that EphA receptors signal via the exchange factor Vav2 to activate RhoA and that both Vav2 and RhoA are required for prostate cancer cell–cell repulsion. Furthermore, we find that in EphA2/EphA4, Vav2 or RhoA siRNA-treated cells, contact repulsion can be restored by partial microtubule destabilisation. We propose that EphA–Vav2–RhoA-mediated repulsion between contacting cancer cells at the tumour edge could enhance their local invasion away from the primary tumour.

show abstract

“…Despite recognition of CIL over half a century ago, methods for its in vitro study have remained largely unchanged, and involve either placing two tissue explants in close proximity in culture, or directly observing rare, serendipitous collisions between dissociated cells [1,3,4,19,20]. As a result, these methods have limited quantitative insight into CIL.…”

Section: Introductionmentioning

confidence: 99%

Contact inhibition of locomotion probabilities drive solitary versus collective cell migration

Desai

Gopal

Chen

et al. 2013

J. R. Soc. Interface.

123

View full text Add to dashboard Cite

Contact inhibition of locomotion (CIL) is the process whereby cells collide, cease migrating in the direction of the collision, and repolarize their migration machinery away from the collision. Quantitative analysis of CIL has remained elusive because cell-to-cell collisions are infrequent in traditional cell culture. Moreover, whereas CIL predicts mutual cell repulsion and 'scattering' of cells, the same cells in vivo are observed to undergo CIL at some developmental times and collective cell migration at others. It remains unclear whether CIL is simply absent during collective cell migration, or if the two processes coexist and are perhaps even related. Here, we used micropatterned stripes of extracellular matrix to restrict cell migration to linear paths such that cells polarized in one of two directions and collisions between cells occurred frequently and consistently, permitting quantitative and unbiased analysis of CIL. Observing repolarization events in different contexts, including headto-head collision, head-to-tail collision, collision with an inert barrier, or no collision, and describing polarization as a two-state transition indicated that CIL occurs probabilistically, and most strongly upon head-to-head collisions. In addition to strong CIL, we also observed 'trains' of cells moving collectively with high persistence that appeared to emerge from single cells. To reconcile these seemingly conflicting observations of CIL and collective cell migration, we constructed an agent-based model to simulate our experiments. Our model quantitatively predicted the emergence of collective migration, and demonstrated the sensitivity of such emergence to the probability of CIL. Thus CIL and collective migration can coexist, and in fact a shift in CIL probabilities may underlie transitions between solitary cell migration and collective cell migration. Taken together, our data demonstrate the emergence of persistently polarized, collective cell movement arising from CIL between colliding cells.

show abstract

The effects of modifying RhoA and Rac1 activities on heterotypic contact inhibition of locomotion

Cited by 14 publications

References 47 publications

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

Calreticulin affects cell adhesiveness through differential phosphorylation of insulin receptor substrate-1

EphA receptors regulate prostate cancer cell dissemination through Vav2–RhoA mediated cell–cell repulsion

Contact inhibition of locomotion probabilities drive solitary versus collective cell migration

Contact Info

Product

Resources

About