alexa Molecular Analysis of RASSF1 Gene Methylation and mRNA Expression in Sporadic Breast Cancer | OMICS International | Abstract
ISSN: 2471-2663

Clinical & Medical Biochemistry
Open Access

Like us on:

OMICS International organises 3000+ Global Conferenceseries Events every year across USA, Europe & Asia with support from 1000 more scientific Societies and Publishes 700+ Open Access Journals which contains over 50000 eminent personalities, reputed scientists as editorial board members.

Open Access Journals gaining more Readers and Citations
700 Journals and 15,000,000 Readers Each Journal is getting 25,000+ Readers

This Readership is 10 times more when compared to other Subscription Journals (Source: Google Analytics)

Research Article

Molecular Analysis of RASSF1 Gene Methylation and mRNA Expression in Sporadic Breast Cancer

Maria Lagiou1, Nikoleta Poumpouridou1, Nikolaos Goutas2, Dimitrios Vlachodimitropoulos2, Aggeliki Pappa1, Evi Lianidou3, Triantafyllos Liloglou4 and Christos Kroupis1*

1Department of Clinical Biochemistry, Attikon University General Hospital, University of Athens Medical School, Haidari 12462, Greece

2Laboratory of Pathologic Anatomy, Evgenidio Hospital, University of Athens Medical School, Athens 15128, Greece

3Laboratory of Analytical Chemistry, Department of Chemistry, University of Athens, Athens 15771, Greece

4Department of Molecular and Clinical Cancer Medicine, University of Liverpool, L3 9TA Liverpool, United Kingdom

*Corresponding Author:
Christos Kroupis
Assistant Professor of Clinical Biochemistry and Molecular Diagnostics
University of Athens Medical School
Attikon University General Hospital
1 Rimini St., Haidari 12462, Greece
Tel: 302105831919
E-mail: [email protected]

Received date: May 02, 2016; Accepted date: May 23, 2016; Published date: May 28, 2016

Citation: Lagiou M, Poumpouridou N, Goutas N, Vlachodimitropoulos D, Pappa A, et al. (2016) Molecular Analysis of RASSF1 Gene Methylation and mRNA Expression in Sporadic Breast Cancer. Clin Med Biochemistry Open Access 2: 118. doi:10.4172/2471-2663.1000118

Copyright: © 2016 Lagiou M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Introduction: The aim of this study was to validate innovative and reliable methods for studying DNA methylation of the first promoter of RASSF1 gene (where its primary transcript RASSF1A in breast tissue is transcribed from), its mRNA expression and their evaluation in sporadic breast cancer. Materials and methods: DNA and RNA from 81 frozen breast cancer tissues with known histopathological data as well as 4 normal breast tissues were analyzed. DNA methylation levels were assessed by Pyrosequencing, analyzing 9 CpG dinucleotides in the CpG island of the first RASSF1 promoter. For mRNA expression, a real-time RT-qPCR method was used and validated with synthetic standards, with the SYBR Green PCR kit and a suitable set of standardized primers (Qiagen) for all RASSF1 transcript variants (except G). For relative quantification of RASSF1 expression, the 2-ΔCt method was used with beta2-microglobulin as a reference gene. Results: 59 samples were characterized as RASSF1 normally-methylated (72.8%), while 22 samples as hypermethylated (27.2%). Also, 40 samples were classified as RASSF1 mRNA over-expressing (49.4%), while 41 (50.6%) as sub-expressing. No inverse correlation was found between methylation and RASSF1 expression (p=0.207). Logistic regression showed that the probability of lymph node infiltration was increased by the presence of negative ER receptors (p=0.008, OR 0.09, CI 0.14-0.52), the percentage of methylation (p=0.006, OR 7.96, CI 1.82-34.86) and the level of RASSF1 expression (p=0.047, OR 3.94, CI 1.02-15.29). Survival analysis showed that there is a marginal statistically significant correlation between metastasis and mRNA RASSF1 over-expression (log rank test, p=0.040). Conclusions: The evaluated assays appear to provide potentially useful information for the clinical management of breast carcinomas. A similar reliable assay for the methylation of the second RASSF1 promoter should be evaluated in the future. Regarding the expression study, a new design strategy with probes specific for all different RASSF1 transcripts, would provide additional power in the study and potentiate the use of new RASSF1 biomarkers in the clinical evaluation of sporadic breast cancer.

Keywords

Peer Reviewed Journals
 
Make the best use of Scientific Research and information from our 700 + peer reviewed, Open Access Journals
International Conferences 2018-19
 
Meet Inspiring Speakers and Experts at our 3000+ Global Annual Meetings

Contact Us

Agri & Aquaculture Journals

Dr. Krish

[email protected]

+1-702-714-7001Extn: 9040

Biochemistry Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Business & Management Journals

Ronald

[email protected]

1-702-714-7001Extn: 9042

Chemistry Journals

Gabriel Shaw

[email protected]

1-702-714-7001Extn: 9040

Clinical Journals

Datta A

[email protected]

1-702-714-7001Extn: 9037

Engineering Journals

James Franklin

[email protected]

1-702-714-7001Extn: 9042

Food & Nutrition Journals

Katie Wilson

[email protected]

1-702-714-7001Extn: 9042

General Science

Andrea Jason

[email protected]

1-702-714-7001Extn: 9043

Genetics & Molecular Biology Journals

Anna Melissa

[email protected]

1-702-714-7001Extn: 9006

Immunology & Microbiology Journals

David Gorantl

[email protected]

1-702-714-7001Extn: 9014

Materials Science Journals

Rachle Green

[email protected]

1-702-714-7001Extn: 9039

Nursing & Health Care Journals

Stephanie Skinner

[email protected]

1-702-714-7001Extn: 9039

Medical Journals

Nimmi Anna

[email protected]

1-702-714-7001Extn: 9038

Neuroscience & Psychology Journals

Nathan T

[email protected]

1-702-714-7001Extn: 9041

Pharmaceutical Sciences Journals

Ann Jose

[email protected]

1-702-714-7001Extn: 9007

Social & Political Science Journals

Steve Harry

[email protected]

1-702-714-7001Extn: 9042

 
© 2008- 2018 OMICS International - Open Access Publisher. Best viewed in Mozilla Firefox | Google Chrome | Above IE 7.0 version
Leave Your Message 24x7