IADR Abstract Archives
Intermediate Filament Impairment Affects Cell Functions Independent From Keratinocyte Transformation
Objectives: Cytoplasmic intermediate filaments (cIFs) play a vital role in maintaining cell physiology, yet the molecular and functional impacts of cIF disturbances remain poorly understood. Key challenges include identifying defects in tissue homeostasis regulated by cell functions and unraveling the complex interrelationships between specific cIFs and distinct cellular processes.
Methods: To investigate the molecular and functional consequences of these early EMT-related changes, RNA interference (RNAi) was used for impairment of key-markers associated with EMT in gingival keratinocytes with different stages of alcohol-induced transformation. We focused on siRNA-treatment targeting Keratin K5 and K14, Vimentin (Vim) and Involucrin (IVL). The effects of RNAi, compared to untreated controls, were analyzed in terms of differentiation, proliferation, nuclear pyknosis, cell morphology and transcriptional activity using techniques such as Western Blot, qPCR, proliferation-assays, light microscopy and indirect immunofluorescence.
Results: Impairment of K5, K14, IVL and VIM has been found to influence proliferation and differentiation. Additionally, disruption of cIFs reduces the expression of nuclear integrity protein lamin B1 and the terminal keratinocyte differentiation marker IVL. Conversely, IVL impairment decreases cIF expression, suggesting a functional regulatory interaction between cIFs and IVL.
Conclusions: In conclusion, these findings highlight that cIF impairment disrupts the balance between proliferation and differentiation, both of which are critical for maintaining tissue homeostasis. Targeted impairment of cIFs provides a valuable approach to exploring their role in cell-dependent tissue physiology at the molecular level and uncovers potential interactions between cIFs and epithelial differentiation. This knowledge extension in the field of epithelial cIF and IVL functions in health and disease is of great benefit, since it can serve to identify and characterize new biomarkers in the context of early cancer detection and prevention.
Methods: To investigate the molecular and functional consequences of these early EMT-related changes, RNA interference (RNAi) was used for impairment of key-markers associated with EMT in gingival keratinocytes with different stages of alcohol-induced transformation. We focused on siRNA-treatment targeting Keratin K5 and K14, Vimentin (Vim) and Involucrin (IVL). The effects of RNAi, compared to untreated controls, were analyzed in terms of differentiation, proliferation, nuclear pyknosis, cell morphology and transcriptional activity using techniques such as Western Blot, qPCR, proliferation-assays, light microscopy and indirect immunofluorescence.
Results: Impairment of K5, K14, IVL and VIM has been found to influence proliferation and differentiation. Additionally, disruption of cIFs reduces the expression of nuclear integrity protein lamin B1 and the terminal keratinocyte differentiation marker IVL. Conversely, IVL impairment decreases cIF expression, suggesting a functional regulatory interaction between cIFs and IVL.
Conclusions: In conclusion, these findings highlight that cIF impairment disrupts the balance between proliferation and differentiation, both of which are critical for maintaining tissue homeostasis. Targeted impairment of cIFs provides a valuable approach to exploring their role in cell-dependent tissue physiology at the molecular level and uncovers potential interactions between cIFs and epithelial differentiation. This knowledge extension in the field of epithelial cIF and IVL functions in health and disease is of great benefit, since it can serve to identify and characterize new biomarkers in the context of early cancer detection and prevention.