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New Rock Inhibitors Action Analysis in the Cytoskeleton and Cell Migration of Tumor Cell Line MDA‐MB 231
Author(s) -
Guerra Fabiana Sélos,
Oliveira Ramon Guerra,
Fraga Carlos Alberto Manssour,
Santos Mermelstein Claudia,
Fernandes Patricia Dias
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.836.7
Subject(s) - rock1 , focal adhesion , rho associated protein kinase , actin cytoskeleton , phalloidin , cell migration , viability assay , cofilin , chemistry , motility , cytoskeleton , kinase , cell , microbiology and biotechnology , biology , biochemistry , protein kinase a
Many patients with metastatic tumors have substantially worse survival in comparison with those patients with nonmetastatic tumors. The ability of cancer cells to undergo invasion and migration is a prerequisite for tumor metastasis and these events became targets for the development of new antimetastatic drugs. Rho kinases, ROCK1 and ROCK2, are key regulators of focal adhesion, actomyosin contraction, and thus cell motility. In this regard our group synthesized two sulfonylhydrazones, analogous to Fasudil (a classic Rho kinase inhibitor), named LASSBio 2020 and LASSBio 2065, differing each other by the presence of one methyl in LASSBio 2065. Therefore, the aim of the present work was to investigate the effects of these analogues against human breast carcinoma (MDA‐MB‐231) cell line migration and signalling pathways involved in in this phenomenon as well as cytoskeleton changes. For the study, cells were incubated with LASSBio 2020 or LASSBio 2065 (0.1 to 30 μM) for 24 or 48 hours. The cell viability was evaluated by MTT (3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐difenltetrazolium) and plasma membrane integrity assay (lactate desidrogenase, LDH). Migration capacity of cells was evaluated using wound healing assay. Morphological analysis was carried out by phase‐contrast micrography, and analysed using Image J software. Actin structures were visualized by fluorescent phalloidin. Statistical analyses were performed by ANOVA with Newman‐Keuls post‐test (*p<0.05). The results showed that none of analogues altered viability of MDA‐MB 231 cells. The effect of the treatment with 10 and 30 μM of LASSBio 2020 decreased cell migration in 30,38% and 32,7%, respectively, when compared to control. LASSBio 2065 decreased cell migration in 24,51% (10 μM) and 26,14% (30 μM), when compared to control. Control cells presented a sprawling appearance with extended cytoplasm and lamelipodios formation. Cells incubated with 10 μm of LASSBio 2020 and LASSBio 2065 presented area, height and width reduced by 40%, membranes with filopodia, protrusions and a fusiform appearance. The staining for F‐actin showed that the cell body became elongated, less spread and with a narrowing of the cell body shape (figure 1). We can conclude that none of analogues (LASSBio 2020 and LASSBio 2065) reduced cell viability. However, the analogues reduced cell migration, altered cell morphology and interfered with formation of actin filaments. Taken together, our data can suggest that the fasudil analogues are a potential inhibitors of metastasis formation. Support or Funding Information Capes, FAPERJ and CNPq This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .