Alterations in expression of S6K1 isoforms in MCF7 cells have a strong impact on the locomotor activity as well as on S6K1 and Akt signaling

Kosach, V. R.; Hotsuliak, N. Ya.; Zaiets, I. V.; Skorokhod, O. M.; Savinska, L. O.; Хоруженко, А. И.; Filonenko, Valeriy

doi:10.7124/bc.000a25

Cited by 4 publications

(10 citation statements)

References 23 publications

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“…Morphological analysis of tumor cells with altered expression of S6K1 isoforms under 2D and 3D culture conditions. Recently we claimed generation of MCF-7 cells with edited by CRISPR/Cas9 technology expression of p60, p70 and p85 S6K1 isoforms: F1 (p85 -/ p70 + /p60 + ), F2 (p85 -/p70 -/p60 + ), and F3 (p85 -/ p70 -/p60 -) [13,14]. It was demonstrated that alterations in the S6K1 isoforms expression differently affects cell migration [14].…”

Section: Resultsmentioning

confidence: 99%

“…Recently we claimed generation of MCF-7 cells with edited by CRISPR/Cas9 technology expression of p60, p70 and p85 S6K1 isoforms: F1 (p85 -/ p70 + /p60 + ), F2 (p85 -/p70 -/p60 + ), and F3 (p85 -/ p70 -/p60 -) [13,14]. It was demonstrated that alterations in the S6K1 isoforms expression differently affects cell migration [14]. Down regulation of the main S6K1 isoforms p70 and p85 with sustained expression of p60 S6K1 dramatically enhances the locomotor activity of MCF-7 cells.…”

Section: Resultsmentioning

confidence: 99%

“…Cell culture. MCF-7 breast cancer cell line and its mutant derivatives MCF-7 p85 -/p70 + /p60 + (clone F1), MCF-7 p85 -/p70 -/p60 + (clone F2), MCF-7 p85 -/p70 -/p60 -(clone F3) [12][13][14] were cultured at 37 °C (5 % CO2) in Dulbecco's Modified Eagle's Medium (DMEM) supple-mented with 10 % fetal bovine serum (FBS, GIBCO), 50 units/ml penicillin and 50 μg/ml streptomycin, 4 mM glutamine.…”

Section: Methodsmentioning

confidence: 99%

“…Although p31S6K1 protein has been shown to lose the kinase domain, it has oncogenic properties [11]. Recently, a novel p60 isoform of S6K1 was found that has the potential of oncogenic properties [12][13][14].…”

Section: Introductionmentioning

confidence: 99%

“…The stable cell lines derived from MCF-7 cells with edited expression of S6K1 isoforms by CRISPR/Cas9 technology were used to evaluate their role in the manifestation of the malignant cell phenotype. Initially we found that down-regulation of the p70 and p85 isoforms expression with sustained expression of p60 S6K1 dramatically enhances the locomotor activity of MCF-7 cells [14]. The migration potential of these cells with direct and indirect influence of fibroblasts was also analyzed.…”

Section: Introductionmentioning

confidence: 99%

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Sensitivity of MCF-7 cells with differential expression of S6K1 isoforms to the regulatory impact of fibroblasts

et al. 2020

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To investigate the involvement of mTOR/S6K1 cell signaling network, with focus on S6K, in response of tumor cells to regulatory impact of fibroblasts. Methods. Cell culture, including co-cultivation of fibroblasts and tumor cells, immunofluorescence analysis, Western blot analysis, assessment of cell migration by scratch test, and transformation of multicellular spheroid in monolayer cell colony. Results. The present work showed the positive effect of stromal cells on the phosphorylation level of the components of mTOR/S6K1 signaling cascade: p85S6K1, p70S6K1 and mTOR in human breast adenocarcinoma MCF-7 cells. To determine, which of the S6K1 isoforms, p85S6K1, p70S6K1 or p60S6K1, is the most sensitive to the extracellular environment, the stable MCF-7 cell lines with edited expression of S6K1 isoforms were used. It was found that selective expression of p60S6K1 led to the changes in morphological features of tumor cells under both two-and three-dimensional culture conditions. These cells also exhibited high levels of focal adhesion kinase (FAK) phosphorylation and large protein content of Zo-1, CD29, CD44 compatible with their high migration potential in the scratch test. Besides, the cells, selectively expressing p60S6K1, were resistant to fibroblastproducing factors and rapamycin. It was also demonstrated that fibroblasts increased tumor cell motility in scratch test and spheroid outspreading assay under co-cultivation conditions in paracrine manner, whereas the direct contact of tumor spheroids with the fibroblast monolayer significantly reduced the velocity of spheroid outspreading. Conclusions. The data obtained indicate that not only the differential expression of S6K1 isoforms in MCF-7 cells but also their ratio are important signaling parameters determining cell survival and response to microenvironment factors.

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Section: Resultsmentioning

confidence: 99%