The biological impact of C11orf30 on transcription regulation during lung epithelial wound healing and epithelial-mesenchymal transition (EMT)

Abstract

Rational: Epithelial-Mesenchymal Transition (EMT), is a highly regulated process that is involved in wound healing and cancer progression. Previous research has shown that when the healing process is dysregulated, EMT can play a role in cancer metastasis. Previous work has shown that EMSY acts as an oncogene in breast and ovarian cancer, while the role that EMSY plays during EMT in lung cancer and the epithelial integrity of lung cells is unknown. This will be addressed in this research project. Objective: To Determine the expression of C11orf30 and the cellular localization of EMSY in response to EMT signals (TGFb1, scratch). Develop a model for the role of EMSY in lung epithelial homeostasis and EMT. Methods & Results: A non-cancerous epithelial cell line, BEAS-2B, as well as a cancerous lung epithelial cell line, A549, were treated for 24 hours with TGF-B1 to induce EMT. Quantitative PCR results showed that during EMT progression, the expression of CDH1 was down-regulated, whereas CDH2 gene expression was increased in bot BEAS-2B and A549 cells. These are both early EMT makers and indicated that early EMT has occurred. CDH1 and CDH2 play a vital role in maintaining epithelial integrity, and dysregulation of these genes could lead to the cells taking on a mesenchymal phenotype. Preliminary results indicated that in untreated BEAS-2B cells, EMSY is located in the nucleus of the cells, where it is able to regulate the gene expression of CDH1. Results also showed that EMSY relocated into the cytoplasm during TGF- B1 induced EMT, in BEAS-2B cells. It was found that the cellular relocation of EMSY, during EMT in BEAS-2B, cells was controlled by TGF-B1 and scratches. QPCR analyses showed that CDH1 gene expression increased in 1% FCS relative to 10% FCS. ChIP analyses revealed that during 1% FCS EMSY binds to the promoter of CDH1. Therefore, it showed that in BEAS2B cells, increased binding of EMSY to CDH1 promoter is correlated with increased CDH1 expression, suggesting EMSY activates CDH1. Conclusion: It was shown that during migration in BEAS-2B cells, EMSY relocates into the nucleus of the cells. Since EMSY is nuclear in untreated BEAS-2B cells it highlights that EMSY must regulate the transcription of genes involved in migration. TGF-B1 induced EMT, in BEAS-2B cells, caused EMSY to be exported out of the nucleus of cells. Therefore, if EMSY is to have a role in EMT, it must involve cellular redistribution. Knowing how EMSY regulates the EMT process in lung cancer cells, expands the data on the functions of EMSY in various cancers. This could lead to the development of novel treatments.

Date of Award	2018
Original language	English