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

• Published in: Biopolimery i kletka 2020, Vol.36 (2), p.110-121
• Main Authors: Kosach, V. R., Hotsuliak, N. Ya, Zaiets, I. V., Skorokhod, O. M., Savinska, L. O., Khoruzhenko, A. I., Filonenko, V. V.
• Format: Article
• Language: English
• Subjects: Actin; AKT protein; Breast cancer; Cell adhesion & migration; Cell lines; Cell migration; Cell morphology; CRISPR; Cytology; Cytoskeleton; Genome editing; Immunofluorescence; Isoforms; Locomotor activity; Morphology; Phosphorylation; Ribosomal protein S6
• ISSN: 0233-7657; 1993-6842
• DOI: 10.7124/bc.000A25

Aim. To generate and characterize MCF-7 cell lines with altered expression of p85, p70 and p60 S6K1 isoforms: p85-/p70-/p60-MCF-7 and p85-/p70-/p60+MCF-7. Methods. CRISPR/Cas9 gene editing, western blot analysis, immunofluorescence analysis, scratch assay. Results. Modified MCF-7 cells with knocked down expression of p85, p70, p60 or only p85 and p70 S6K1 isoforms were generated. Selective inhibition of only p85 and p70 isoforms in p85-/p70-/p60+MCF-7 cellswas accompanied by actin cytoskeleton rearrangements, appearance of fibroblast-like cell morphology and significantly increasedcell locomotor activity. Downregulation of all three S6K1 isoforms in p85-/p70-/p60 – MCF-7 cells inhibited cell migration with no changes in the cell morphology. Alterations inhad a different impact on the ribosomal protein S6 phosphorylation and Akt signaling. Conclusion. Analysis of the modified MCF-7 cell lines revealed different impact of expression of S6K1 isoforms on MCF7 cell locomotor activity and the S6K1– and Akt-dependent signaling. Our data suggest that p60-S6K1 could be involved in regulation of the cell migration. The generated cells can be used for furtheranalysis of functionalactivity of the S6K1 isoforms.