<abstract xml:lang="en" xmlns="http://www.w3.org/1999/xhtml"><p>Genetic alterations and changes in genomic DNA cytosine methylation patterns are associated with all types of cancer and are caused by germline mutations in DNA mismatch repair (MMR) genes, predominantly MLH1 (<italic>MutL homolog 1, 19 exons</italic>) and MSH2 (<italic>MutS homolog 2, 16 exons</italic>). Genomic DNA was extracted from tissue samples embedded in paraffin from 49 patients with adenocarcinoma and from 21 patients with carcinoma for the study group; genomic DNA was extracted from lymphocytes from 10 healthy donors for the control group. We used methylation specific multiplex ligation-dependent probe amplification (MS-ML-PA), which allows the detection of copy number changes and unusual methylation levels of 10 to 50 different sequences in one reaction by use of the methylationsensitive restriction enzyme HhaI and sequence-specific capillary electrophoresis for the study of 24 genes.</p><p>We found the mean methylation rates for MLH1 (97.14%), MSH2 (24.28%), MSH6 (<italic>MutS homolog 6</italic>) (67.14%), MSH3 (<italic>MutS homolog 3</italic>) (78.57%), MLH3 (<italic>MutL homolog 3</italic>) (75.71%), PMS2 (<italic>postmeiotic segregation increased 2</italic>) (65.71%), MGMT(<italic>O-6-methylguanine-DNA methyltransferase</italic>) (82.85%). We conclude that the mismatch repair (MMR) system is critical for the maintenance of genomic stability.</p></abstract>

Genetic alterations and changes in genomic DNA cytosine methylation patterns are associated with all types of cancer and are caused by germline mutations in DNA mismatch repair (MMR) genes, predominantly MLH1 (MutL homolog 1, 19 exons) and MSH2 (MutS homolog 2, 16 exons). Genomic DNA was extracted from tissue samples embedded in paraffin from 49 patients with adenocarcinoma and from 21 patients with carcinoma for the study group; genomic DNA was extracted from lymphocytes from 10 healthy donors for the control group. We used methylation specific multiplex ligation-dependent probe amplification (MS-ML-PA), which allows the detection of copy number changes and unusual methylation levels of 10 to 50 different sequences in one reaction by use of the methylationsensitive restriction enzyme HhaI and sequence-specific capillary electrophoresis for the study of 24 genes.We found the mean methylation rates for MLH1 (97.14%), MSH2 (24.28%), MSH6 (MutS homolog 6) (67.14%), MSH3 (MutS homolog 3) (78.57%), MLH3 (MutL homolog 3) (75.71%), PMS2 (postmeiotic segregation increased 2) (65.71%), MGMT(O-6-methylguanine-DNA methyltransferase) (82.85%). We conclude that the mismatch repair (MMR) system is critical for the maintenance of genomic stability.

Alterations of Copy Number of Methylation Pattern in Mismatch Repair Genes by Methylation Specific-Multiplex Ligation-Dependent Probe Amplification in Cases of Colon Cancer

Department of Medical Genetics, Afyon Kocatepe University of Medical Faculty, Afyonkarahisar, Turkey

Department of Molecular Biology, Afyon Kocatepe University of Science Faculty, Afyonkarahisar, Turkey

Department of Internal Medicine and Gastroenterology, Celal Bayar University of Medicine Medical Faculty, Manisa, Turkey

Güneş Pathology Laboratory, Alsancak, Izmir, Turkey

Balkan Journal of Medical Genetics

Genetic alterations and changes in genomic DNA cytosine methylation patterns are associated with all types of cancer and are caused by germline mutations in...

Alterations of Copy Number of Methylation Pattern in Mismatch Repair Genes by Methylation Specific-Multiplex Ligation-Dependent Probe Amplification in Cases of Colon Cancer

Published Online: Dec 08, 2011

Page range: 25 - 34

DOI: https://doi.org/10.2478/v10034-011-0044-x

Keywords
Colon cancer, Methylation specificmultiplex ligation-dependent probe amplification (MS-MLPA), Mismatched repair (MMR) Genes, Methylation, Copy number

This content is open access.

Alterations of Copy Number of Methylation Pattern in Mismatch Repair Genes by Methylation Specific-Multiplex Ligation-Dependent Probe Amplification in Cases of Colon Cancer

Published Online: Dec 08, 2011

Page range: 25 - 34

DOI: https://doi.org/10.2478/v10034-011-0044-x

KeywordsColon cancer, Methylation specificmultiplex ligation-dependent probe amplification (MS-MLPA), Mismatched repair (MMR) Genes, Methylation, Copy number

This content is open access.

Keywords
Colon cancer, Methylation specificmultiplex ligation-dependent probe amplification (MS-MLPA), Mismatched repair (MMR) Genes, Methylation, Copy number