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Cancer-associated mutations in the protrusion-targeting region of p190RhoGAP impact tumor cell migration

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ABSTRACT

p190RhoGAP (p190A) is a negative regulator of RhoA and localizes to membrane protrusions, where its GAP activity is required for directional migration. Here, Binamé et al. identify the protrusion-localization sequence in p190A and show that cancer-associated mutations in this region affect p190A localization and function as well as tumor cell migration.

No MeSH data available.


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Mutations in PLS affect MDA-MB-231 directed cell migration. MDA-MB-231 cells were cotransfected with p190A- and EGFP-expressing constructs, spread on gelatin coating, and visualized by epifluorescence and contrast time-lapse microscopy for 16 h. Frames were collected every 10 min. (A) Representative tracings of paths traveled over 16 h by MDA-MB-231 cells transfected with p190A mutants. (B) Quantification of movement directionality represented as a ratio of the distance between the start and end point over the total path traveled. (C) Cell velocity is calculated by length of the path traveled divided by time. Values are expressed as the mean ± SEM (n = 20 cells per condition). Statistical significance was calculated relative to nontransfected (NT) condition. *, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, not significant.
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fig8: Mutations in PLS affect MDA-MB-231 directed cell migration. MDA-MB-231 cells were cotransfected with p190A- and EGFP-expressing constructs, spread on gelatin coating, and visualized by epifluorescence and contrast time-lapse microscopy for 16 h. Frames were collected every 10 min. (A) Representative tracings of paths traveled over 16 h by MDA-MB-231 cells transfected with p190A mutants. (B) Quantification of movement directionality represented as a ratio of the distance between the start and end point over the total path traveled. (C) Cell velocity is calculated by length of the path traveled divided by time. Values are expressed as the mean ± SEM (n = 20 cells per condition). Statistical significance was calculated relative to nontransfected (NT) condition. *, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, not significant.

Mentions: Finally, we tested the impact of these p190A mutations on cancer cell migration using MDA-MB-231 breast carcinoma cells plated on gelatin-coated dishes. Time-lapse microscopy experiments were performed after cotransfection of the cells with p190A- and EGFP-expressing constructs (Videos 1–5). Analysis of the transfected cell paths revealed that the directionality of the cells expressing p190A mutants is altered compared with their WT counterpart (Fig. 8, A and B), whereas the speed of migration remains unaffected (Fig. 8 C). Thus, this result demonstrates that the S866F and Δ(865–870) cancer-associated mutations may favor turning behavior, which reflects exploration of cancerous cells.


Cancer-associated mutations in the protrusion-targeting region of p190RhoGAP impact tumor cell migration
Mutations in PLS affect MDA-MB-231 directed cell migration. MDA-MB-231 cells were cotransfected with p190A- and EGFP-expressing constructs, spread on gelatin coating, and visualized by epifluorescence and contrast time-lapse microscopy for 16 h. Frames were collected every 10 min. (A) Representative tracings of paths traveled over 16 h by MDA-MB-231 cells transfected with p190A mutants. (B) Quantification of movement directionality represented as a ratio of the distance between the start and end point over the total path traveled. (C) Cell velocity is calculated by length of the path traveled divided by time. Values are expressed as the mean ± SEM (n = 20 cells per condition). Statistical significance was calculated relative to nontransfected (NT) condition. *, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, not significant.
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Related In: Results  -  Collection

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fig8: Mutations in PLS affect MDA-MB-231 directed cell migration. MDA-MB-231 cells were cotransfected with p190A- and EGFP-expressing constructs, spread on gelatin coating, and visualized by epifluorescence and contrast time-lapse microscopy for 16 h. Frames were collected every 10 min. (A) Representative tracings of paths traveled over 16 h by MDA-MB-231 cells transfected with p190A mutants. (B) Quantification of movement directionality represented as a ratio of the distance between the start and end point over the total path traveled. (C) Cell velocity is calculated by length of the path traveled divided by time. Values are expressed as the mean ± SEM (n = 20 cells per condition). Statistical significance was calculated relative to nontransfected (NT) condition. *, P < 0.05; **, P < 0.01; ***, P < 0.001; n.s, not significant.
Mentions: Finally, we tested the impact of these p190A mutations on cancer cell migration using MDA-MB-231 breast carcinoma cells plated on gelatin-coated dishes. Time-lapse microscopy experiments were performed after cotransfection of the cells with p190A- and EGFP-expressing constructs (Videos 1–5). Analysis of the transfected cell paths revealed that the directionality of the cells expressing p190A mutants is altered compared with their WT counterpart (Fig. 8, A and B), whereas the speed of migration remains unaffected (Fig. 8 C). Thus, this result demonstrates that the S866F and Δ(865–870) cancer-associated mutations may favor turning behavior, which reflects exploration of cancerous cells.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

p190RhoGAP (p190A) is a negative regulator of RhoA and localizes to membrane protrusions, where its GAP activity is required for directional migration. Here, Binam&eacute; et al. identify the protrusion-localization sequence in p190A and show that cancer-associated mutations in this region affect p190A localization and function as well as tumor cell migration.

No MeSH data available.


Related in: MedlinePlus