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Institute for Biological Research "Sinisa Stankovic"
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Uloga metilacije DNK u procesu epitelno-mezenhimske tranzicije humanih epitelnih ćelija konjuktive

The role of DNA methylation in the process of epithelial-mesenchymal transition of human conjunctival epithelial cells

2020
Full text (6.540Mb)
Authors:
Rajić, Jovana
Mentors:
Grdović, Nevena
Vidaković, Melita
Document Type:
Doctoral thesis (Published version)
,
© 2020 by the author
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Abstract:
The process of epithelial-mesenchymal transition (EMT) could be an important factor in development of fibrosis-related conjunctival eye diseases, but its precise role in these conditions has not been defined yet. The aim of this doctoral dissertation was to examine the ability of human epithelial conjunctival cell line (HCjE) to enter EMT process after infection with trachoma causative agent, bacteria Chlamydia trachomatis, i to identify key factors associated with the initiation i progression of transition in TGF-β-induced EMT model. Special focus was directed to elucidating the role of DNA methylation in the regulation of expression of genes involved in the transition of HCjE cells. Activation of signaling pathways associated with the initiation of the EMT process i the expression level of epithelial/mesenchymal markers revealed that HCjE cells possess the ability to enter the transition after infection with bacteria C. trachomatis, which was associated with changes in the methylation profiles of epithelial/mesenchymal gene markers. Cell morphology, migration ability i expression level of marker genes indicated that long-term treatment with TGF-β1 isoform has the greatest potential to initiate EMT process in HCjE cells, while treatments with DNA demethylating agent suggested that DNA methylation is an important mechanism which underlies this process. Changes in DNA methylation profile, detected by melting curve analysis i bisulfite sequencing, highlighted a key role of miR-200 family members in initiating i reversing TGF-β1-induced EMT process in HCjE cells. The results obtained in this doctoral dissertation indicate the possibility of developing new selective therapeutic strategies in the treatment of fibrosis-related conjunctival diseases, based on the reversibility of both EMT process i epigenetic mechanisms.
Keywords:
Epithelial-mesenchymal transition (EMT); Conjunctival fibrosis; Human epithelial conjunctival cells (HCjE); Chlamydia trachomatis; TGF-β; DNA methylation; 5-Azacytidine (5-AzaC); E-cadherin (CDH1); miRNA; miR-200 family
Source:
Faculty of Biology, University of Belgrade, 2020, 1-133
Projects:
  • Signaling molecules in diabetes: search for potential targets in intrinsic pathways for prediction and intervention in diabetes (RS-173020)
  • Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200007 (University of Belgrade, Institute for Biological Research 'Siniša Stanković') (RS-200007)
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URI
https://radar.ibiss.bg.ac.rs/123456789/3884
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  • Doctoral Thesis
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Vancouver:
Rajić J. The role of DNA methylation in the process of epithelial-mesenchymal transition of human conjunctival epithelial cells [dissertation]. Belgrade: Faculty of Biology, University of Belgrade; 2020. 133 p.

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