IADR Abstract Archives
E-cadherin Inhibition Effects on Oral Squamous Cell Carcinoma Metastatic Genes
Objectives: Oral Squamous Cell Carcinoma (OSCC) is an invasive multistage process malignancy that affects the oral stratified squamous epithelial cells. It is associated with high death rates worldwide. E-cadherin protein is an important marker of epithelial tissue and it is one of the key regulators of tissue integrity and polarity. Generally, the effect of E-cadherin protein down regulation in tumour progression is well known to affect overall prognosis of cancer pateints. This down regulation might have a role in metastatic process of the cancer which require assessing the Epithelial-Mesenchymal Transition genes (EMT) that intiate this process.
Methods: Mouse Embryonic Stem cells (mESCs) were used as a model of epithelial tissue to study the effect of E-cadherin inhibition in regulation of EMT genes and compared to E-cadherin knock out mESCs (E-cad-/- mESCs). Microarray analysis and Chromatin immunoprecipitation ChIP-seq were done and results were assessed using Cytoscape Network modelling programme (ModuLand) and the UCSC genome browser respectively. Methylation sequencing was done for these cells to assess the the methylated promotor areas for genes of interest.
Results: The Microarray has shown several thousand transcripts alterations which govern a number of biological processes and Histone acetyltransferases EP300 was identified as putative key central regulatory factor in network modelling. Therefore, EP300 and Histone H3 (H3K27ac) antibodies ChIP-seq was done to determine how transcription factors influence phenotype-affecting mechanisms. Methylation sequencing revealed a significant difference between mESCs and Ecad-/-mESCs.
Conclusions: The inhibition of E-cadherin protein causes wide alteration of some EMT genes and changing of the binding sites of EP300 and H3K27ac to these genes between mESC and E-cad-/- mESC. In addition, this inhibition affects the methylation of promotor areas of some EMT genes. The key targets from microarray, ChIP-seq and methylation sequencing need to be further validated and investigated on mESC model and OSCC tissue sections.
Methods: Mouse Embryonic Stem cells (mESCs) were used as a model of epithelial tissue to study the effect of E-cadherin inhibition in regulation of EMT genes and compared to E-cadherin knock out mESCs (E-cad-/- mESCs). Microarray analysis and Chromatin immunoprecipitation ChIP-seq were done and results were assessed using Cytoscape Network modelling programme (ModuLand) and the UCSC genome browser respectively. Methylation sequencing was done for these cells to assess the the methylated promotor areas for genes of interest.
Results: The Microarray has shown several thousand transcripts alterations which govern a number of biological processes and Histone acetyltransferases EP300 was identified as putative key central regulatory factor in network modelling. Therefore, EP300 and Histone H3 (H3K27ac) antibodies ChIP-seq was done to determine how transcription factors influence phenotype-affecting mechanisms. Methylation sequencing revealed a significant difference between mESCs and Ecad-/-mESCs.
Conclusions: The inhibition of E-cadherin protein causes wide alteration of some EMT genes and changing of the binding sites of EP300 and H3K27ac to these genes between mESC and E-cad-/- mESC. In addition, this inhibition affects the methylation of promotor areas of some EMT genes. The key targets from microarray, ChIP-seq and methylation sequencing need to be further validated and investigated on mESC model and OSCC tissue sections.