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The Regulation of Plasma Gelsolin by DNA Methylation in Ovarian Cancer ChemoresistanceManzoor, Hafiza Bushra 20 September 2023 (has links)
Ovarian cancer (OVCA) is the most lethal gynecologic cancer. Chemoresistance remains a major hurdle to successful therapy and patient survival. The secreted isoform of the actin-associated protein, gelsolin (plasma gelsolin; pGSN), is highly expressed in chemoresistant than chemosensitive OVCA cells, although the mechanism underlying the differential expression is not known. Also, its overexpression significantly correlates with shortened survival of OVCA patients. DNA methylation plays a key role in the regulation of genes expression and contributing to cancer development and chemoresistance with the help of DNA methyltransferases (DNMTs) or Ten eleven translocation (TETs) enzymes. TET1 is the most studied isoform of TETs family and primarily responsible for 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) oxidation to initiate demethylation and increase in the expression of methylated genes. Whether pGSN expression in OVCA cells is regulated by DNA methylation and TET1 regulates the differential pGSN expression between chemosensitive and resistant OVCA cells is not known. In this study, we hypothesized pGSN overexpression in chemoresistant OVCA cells is due to the hypomethylation at its promoter region by TET1. Our objective was to investigate whether DNA methylation and specifically TET1 plays a role in the regulation of differential pGSN expression and chemosensitivity in OVCA. Chemosensitive and resistant OVCA cell lines of different histological subtypes were used in this study to measure pGSN and TET1 mRNA abundance and protein contents by qPCR and Western blotting respectively. Cisplatin-induced chemoresponsiveness was morphologically assessed by Hoechst staining (apoptosis). Infinium HumanMethylation450 BeadChip assay was used for global methylation analysis of twelve (12) different OVCA cells and to investigate the role of DNA methylation specifically in pGSN regulation and pGSN-induced chemoresistance. DNMTs and TETs were pharmacologically inhibited in sensitive and resistant OVCA cell using specific inhibitors. Gain-and-loss-of-function assays were carried to identify the relationship between TET1 and pGSN in OVCA chemoresponsiveness. Differential protein and mRNA expressions of pGSN and TET1 were observed between sensitive and resistant OVCA cells and cisplatin reduced their expression in sensitive but not in resistant cells. Global methylation analysis revealed hypomethylation in resistant cells compared to sensitive cells. Pharmacological inhibition of DNMTs increased pGSN protein levels in sensitive OVCA cells and decreases their responsiveness to cisplatin, however we did not observe any difference in methylation level at pGSN promoter region. TETs inhibition resulted in hypermethylation at multiple CpG sites and decreased pGSN protein level in resistant OVCA cells which was also associated with enhanced response to cisplatin, findings that suggested the methylation role of TETs in the regulation of pGSN expression in OVCA cells. Further, we found that TET1 is inversely related to pGSN and positively related to chemoresponsiveness of OVCA cells. This project does not only broaden our knowledge about the mechanistic insights into the epigenetic regulation of pGSN in OVCA chemoresistance, but it also reveals a new potential target to re-sensitize chemotherapy resistant OVCA cells. This may provide a future strategy to improve overall OVCA patient survival.
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The role of Ten Eleven Translocation enzymes in the hair follicle mesenchymeAhmed, Aqib January 2022 (has links)
Epigenetic mechanisms play an important role during the morphogenesis
of the hair follicle and the hair cycle. Work on hair regeneration is of
importance as no products are available which can provide complete
reversal of hair loss. Tet2 promotes DNA demethylation by the
hydroxylation of 5mC to 5hmC which in turn causes gene transcription
activation. Dermal papilla (DP) cells located within the hair follicle are
responsible for the regulation of development and the growth of hair
follicles. Fgf20 signalling controls commitment of the mesenchymal
precursor cells to the DP progenitor lineage. An immature DP cells is then
formed during maturation by Shh signalling which then stimulates these to
differentiate into a DP cell by BMP and Wnt signalling.
Methylated DNA can be bound by the proteins recruiting transcription corepressors.
DNA methyltransferases (DNMT’s) can be degraded by
decitabine which reverses gene silencing. Conditional knockout of Tet2 in
mouse DP cells results in a delay in anagen initiation, suggesting Tet2 is
involved in the telogen-anagen transition. Additionally, by using dermal
fibroblasts and RA-DPAC (Dermal Papilla activating medium
supplemented with retinoic acid), it was found that decitabine can increase
plasticity in dermal fibroblasts and RA-DPAC can be used to accelerate a
lineage change to DP cells which is supported by the significant increase in the DP specific gene expression. Examples include AlPl, LEF1,
BMP4/6/7, FGF10, BMPR1A and PDGFA. Additionally, by way of siRNA
and conditional Tet2 knockout data in dermal fibroblasts, it was found Tet2
regulates signature DP genes such as Bmpr1a, ALPL, Tcf4 and SOX2.
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Mechanisms of Fgf8 transcription in the developing mouse olfactory placode.LINSCOTT, MEGAN L. 20 April 2020 (has links)
No description available.
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