Mechanical profile of human keratinocytes expressing HPV-18 oncogenes

• Published in: Biochemical and biophysical research communications 2023-05, Vol.657, p.86-91
• Main Authors: Millán, Magdalena, Villarreal, Lihuén, D'Aiuto, Natali, Bologna-Molina, Ronell, Sotelo-Silveira, José, Benech, Juan C., Hochmann, Jimena, Arocena, Miguel
• Format: Article
• Language: English
• Subjects: AFM; Cell mechanics; Cell Transformation, Neoplastic - genetics; Female; HaCaT E5/E6/E7-18; Human papillomavirus 18 - genetics; Humans; Keratinocytes - metabolism; Oncogene Proteins, Viral - genetics; Oncogene Proteins, Viral - metabolism; Oncogenes; Papillomavirus E7 Proteins - metabolism; Pre-tumoral model; Repressor Proteins - metabolism
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2023.03.054

During tumorigenesis, the mechanical properties of cancer cells change markedly, with decreased stiffness often accompanying a more invasive phenotype. Less is known about the changes in mechanical parameters at intermediate stages in the process of malignant transformation. We have recently developed a pre-tumoral cell model by stably transducing the immortalized but non-tumorigenic human keratinocyte cell line HaCaT with the E5, E6 and E7 oncogenes from HPV-18, one of the leading causes of cervical cancer and other types of cancer worldwide. We have used atomic force microscopy (AFM) to measure cell stiffness and to obtain mechanical maps of parental HaCaT and HaCaT E5/E6/E7-18 cell lines. We observed a significant decrease in Young's modulus in HaCaT E5/E6/E7-18 cells measured by nanoindentation in the central region, as well as decreased cell rigidity in regions of cell-cell contact measured by Peakforce Quantitative Nanomechanical Mapping (PF-QNM). As a morphological correlate, HaCaT E5/E6/E7-18 cells displayed a significantly rounder cell shape than parental HaCaT cells. Our results therefore show that decreased stiffness with concomitant perturbations in cell shape are early mechanical and morphological changes during the process of malignant transformation. •A characterization of the mechanical properties of keratinocytes expressing HPV-18 oncogenes was performed.•Cells expressing HPV-18 oncogenes had decreased stiffness and rounder shape compared to parental cells.•Our results show that mechanical and morphological alterations occur early in the process of malignant transformation.