2022 / 06

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DOI: 10.24075/brsmu.2022.070

ORIGINAL RESEARCH

Knockout of mutant TP53 in the HaCaT cells enhances their migratory activity

Kozhin PM, Romashin DD, Rusanov AL, Luzgina NG
About authors

Orekhovich Research Institute of Biomedical Chemistry, Moscow, Russia

Correspondence should be addressed: Alexander L. Rusanov
Pogodinskaya, 10, str. 8, Moscow, 119121, Russia; moc.liamg@vonasur.l.rednaxela

About paper

Funding: the study was carried out as part of the Program for fundamental research in the Russian Federation for the long-term period (2021–2030) (№ 122022800481-0).

Author contribution: Luzgina NG, Rusanov AL — study concept; Romashin DD, Kozhin PM, Luzgina NG, Rusanov AL — study design and literature review; Romashin DD, Kozhin PM — study planning and execution; Kozhin PM, Romashin DD, Luzgina NG, Rusanov AL — data analysis and interpretation; Kozhin PM, Romashin DD — manuscript writing; Kozhin PM, Romashin DD, Luzgina NG, Rusanov AL — manuscript editing.

Compliance with ethical standards: the study was carried out in accordance with the Declaration of Helsinki of the World Medical Association and all its revisions.

Received: 2022-12-12 Accepted: 2022-12-24 Published online: 2022-12-30
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Fig. 1. Proliferative activity of HaCaT cells. А. Distribution of wild type (WT) HaCaT cells and mutTP53 knockout (dp53) HaCaT cells by cell cycle phases. B. Proliferative activity of wild type (WT) HaCaT cells and mutTP53 knockout (dp53) HaCaT cells
Fig. 2. Scratch test. Rates of monolayer defect closure by various cell lines: normal keratinocytes (NHK), wild type (WT) HaCaT cells, and mutTP53 knockout (dp53) HaCaT cells. A. Light microscopy; magnification ×40; day 0 (immediately after injury) and day 3 after scratching. B. Generalized chart of the relative defect area observed on days 1, 2, 3 after scratching. C. Relative expression of genes LAMB3, LAMC2. * — significant differences compared to wild type HaCaT (p < 0.05); # — significant differences compared to NHK on the corresponding day (p < 0.05)
Fig. 3. Multilayered skin equivalent formation by keratinocytes of different lines: normal keratinocytes (NHK), wild type (WT) HaCaT cells, and mutTP53 knockout (dp53) HaCaT cells. А. Immunofluorescence microscopy. Stain: nuclei (DAPI) — blue, KRT5 — red; magnification ×400. B. Transepithelial electrical resistance (TEER) values
Table 1. Primers used during the study