P21 ELF5 is required to prevent keratinocyte malignancy transformation in vitro
Chandra Kala Khadka, Mohammed AhmedAbstract
Introduction and aims
Skin being the largest organ in the body has three layers: epidermis, dermis and hypodermis. The epidermal layer itself has multiple layers with self-renewal capabilities. Malignant transformation of keratinocyte carcinoma occurs owing to failure in the completion of differentiation programme, which can lead to dysplastic epithelium and precancerous cells in skin. E74-like factor (ELF)5 is a member of the E26 transformation-specific (ETS) family of transcription factors, predominantly expressed in some epithelial tissues including skin and is known to have an antitumorigenic role in other epithelial tissues. We have shown that ELF5 is essential for normal keratinocytes proliferation, differentiation and stem/progenitor maintenance in mouse skin and loss of ELF5 in human immortalized keratinocytes can lead to tumorigenesis. Further analysis is required to clearly define the role of ELF5 in malignant keratinocyte development.
Methods
Our investigations provide novel approach into the role of ELF5 in keratinocytes activities and its malignant transformation. Using cancer cell lines such as SCC-13, A431 and SCC-9, we transduced cells lentiviral particles (ELF5 control and ELF5 overexpression) with subsequent Fluorescence-activated cell sorting live cell sorting, followed by transcriptomic (RNA sequencing analysis) and proteomic analyses (quantitative polymerase chain reaction, Western blot analysis) and functional studies (migration transwell assay, soft agar and three-dimensional colony-forming assay) in vitro.
Results
Our findings suggest that overexpression of ELF5 is instrumental in reducing migrations and inhibiting skin tumorigenesis, thus representing a potential target for future therapeutic interventions in malignant.
Conclusions
Our data demonstrate an important understanding of ELF5 as a crucial regulator of keratinocyte malignant transformation and will provide better understanding of the novel transcriptional regulator in skin in many areas of research, including stem cell and cancer biology.