Κατεύθυνση Κλινική Χημεία
Library of the School of Science

2014-10-27

2014

Μαστοράκη Σοφία

Ε. Λιανίδου Καθηγήτρια Ε.Κ.Π.Α (επιβλέπουσα), Γ. Σωτηροπούλου Αναπλ. Καθηγήτρια Πανεπιστημίου Πατρών, Α. Ψυρρή Επίκ. Καθηγήτρια Ε.Κ.Π.Α

Μελέτη και κλινική αξιολόγηση της μεθυλίωσης DNA των υποκινητών των γονιδίων SOX17 και BRMS1 στον καρκίνο του μαστού με υψηλής διακριτικής ικανότητας ανάλυση καμπυλών τήξης (MS-HRMA)

Greek

Study and clinical evaluation of SOX17 and BRMS1 promoter methylation in breast cancer patients using Methylation-Sensitive-High Resolution Melting Analysis (MS-HRMA)

Aberrant DNA methylation is common in many human cancers, and is an early event
in tumor development and progression. It is one of the major factors for
epigenetic silencing of certain genes, through promoter hypermethylation of
tumor suppressor genes. As a result, the evaluation of promoter methylation of
specific genes could offer important diagnostic or prognostic information on
the clinical outcome of cancer patients. SOX17 and BRMS1 genes, featuring a CpG
island within their promoters, are frequently methylated in that region in
cancer cells. Methylation-sensitive high resolution melting (MS-HRMA) enables
highly sensitive, labor- and cost-efficient single locus methylation studies on
the basis of DNA high-resolution melting analysis technology (HRMA). The aim of
this Diploma thesis, was the development and clinical evaluation of a novel
MS-HRMA method based on the combination of Real-Time PCR and high-resolution
melting analysis for the investigation of the methylation status of SOX17 and
BRMS1 genes in breast tissues (FFPEs), and the comparison of the results
obtained for both genes, with the respective results of Real-Time MSP. The
developed methodology was extensively analytically optimized and validated and
then applied in 165 FFPE breast tissue samples (134 breast tumors obtained from
patients with early breast cancer and metastasis verified, 15 normal breast
tissues and 16 DNA samples isolated from white blood cells) for SOX17 as well
as in 148 FFPE breast tissue samples (117 breast tumors, 15 normal FFPEs and 16
DNA samples isolated from white blood cells) for BRMS1. As far as the SOX17
gene is concerned, methylation in various levels was observed in 96 out of the
134 tumor samples (71.6%), while none of the non cancerous tissues and samples
were found to be methylated. 15 out of 117 (12.8%) samples investigated for the
BRMS1 gene were found to be methylated. The results obtained by MS-HRMA for the
same samples show a strong correlation with MSP when investigating SOX17
promoter methylation status. The developed methodology can not only, detect the
presence of methylation within the studied region, but also gives a
semi-quantitative estimation of the methylation level in terms of percentage as
well.

DNA methylation, MS-HRMA, Breast cancer, SOX17, BRMS1

Yes

VIII-XIV

Yes

171

XVII, 159

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https://pergamos.lib.uoa.gr/uoa/dl/object/1318425