Evaluation ofLATS1andLATS2Promoter Methylation with the Risk of Pterygium Formation

Purpose. Pterygium is a serious eye problem in countries with high exposure to UV. However, despite numerous studies, the molecular etiology of pterygium is unclear. Recent studies have indicated thatLATS1andLATS2genes are involved in DDR signaling pathways against continuous UV exposure. Our aim was to evaluate theLATS1andLATS2promoter methylation with the risk of pterygium formation.Methods. We evaluated the promoter methylation status ofLATS1andLATS2using methylation-specific PCR technique. Also, mRNA expression ofLATS1andLATS2was assessed in 14 cases of pterygium and 14 normal specimens by real-time PCR.Results. Promoter methylation ofLATS1andLATS2was detected significantly between pterygium tissues and normal tissues [LATS1; OR = 4.9; 95% CI: 1.54 to 15.48,P=0.003;LATS2; OR = 7.1; 95% CI: 1.53 to 33.19,P=0.004]. The gene expression analysis showed a statistically significant difference between pterygium tissues and healthy controls for bothLATS1andLATS2(P<0.05).Conclusions. The data of this study is the first report regarding the effect of promoter methylation of theLATS1andLATS2in the pterygium. To confirm these data, doing further studies in various genetic populations with large sample sizes using advanced molecular techniques is proposed.

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Molecular Genetics of Esophageal Cancer : Indian Perspective

Asian Pacific Journal of Cancer Biology ◽

10.31557/apjcb.2021.6.2.155-160 ◽

2021 ◽

Vol 6 (2) ◽

pp. 155-160

Author(s):

Suddhasattwa Ray ◽

Mona Malekzadehmoghani ◽

Sonia S Ray ◽

Partha Sen ◽

Sayan Chakraborty

Keyword(s):

Promoter Methylation ◽

Methylation Status ◽

Cpg Islands ◽

Stage Iv ◽

Tissue Samples ◽

Cpg Dinucleotides ◽

Specific Pcr ◽

Significant Difference ◽

Indian Perspective

Background: RIZ1 is one of the tumor-suppressor genes that is silenced in many human cancers. Change in RIZ1 expression has not been reported in ESCC patients. Therefore, the aim of this study was to investigate the role of RIZ1 in ESCC in the Indian population. Methods: Twelve esophageal squamous-cell carcinoma (ESCC) patients in stage IV and 12 healthy individuals were used in this study. Tissue sampling was taken from individuals and total RNA was isolated and then cDNA was synthesized using PCR. RIZ1 primers were then designed, and RIZ1 expression was quantified by qRT-PCR. Mapping of CpG islands in RIZ1 promoter was performed using bioinformatics tools. The promoter methylation status of this gene was studied using u methylation-specific PCR (MSP). T-student test was used to analyze the data.Results: Decreased RIZ1 expression was observed in ESCC compared with healthy controls. The results showed a relatively higher density of CpG dinucleotides in the RIZ1 promoter. No significant difference in promoter methylation was observed in blood and tissue samples.Conclusion: The study showed a significant down-regulation of RIZ-1 gene in the blood and tissue samples of ESCC patients that did not related to the altered promoter methylation.

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The Clinical Significance of Promoter Methylation of Fluoropyrimidine Metabolizing and Cyclooxygenase Genes in Colorectal Cancer

Epigenetics Insights ◽

10.1177/2516865720986231 ◽

2021 ◽

Vol 14 ◽

pp. 251686572098623

Author(s):

Mariam Ahmed Fouad ◽

Salem Eid Salem ◽

Marwa M. Hussien ◽

Doaa Mohamed Badr ◽

Abdelrahman N. Zekri ◽

...

Keyword(s):

Colorectal Cancer ◽

Promoter Methylation ◽

Dihydropyrimidine Dehydrogenase ◽

Methylation Status ◽

Pairwise Comparisons ◽

Specific Pcr ◽

Significant Difference ◽

High Level ◽

The Impact ◽

Cox2 Expression

Aims: This study investigated the impact of promoter methylation of flouropyrimidine (FP) metabolizing and cyclooxygenase 2 (COX2) genes on their mRNA expression and on the clinical outcome of colorectal cancer (CRC) patients. Methods: Methylation specific-PCR and real time-PCR of thymidylate synthase (TS), thymidine phosphorylase (TP), dihydropyrimidine dehydrogenase (DPD) and COX2 were performed at baseline and after 3 and 6 months of FP therapy. Pairwise comparisons were conducted between the subgroups of CRC patients. The event free survival (EFS) and the hazard of progression were estimated by univariate and multivariate analyses. Results: At baseline CRC patients, both TS and TP were overexpressed, in spite of the unmethylation of TS and the full methylation of TP genes. Significant downexpression of DPD and COX2 were associated their promoter’s methylation. At the end of FP therapy, TS, DPD and COX2 were overexpressed by 7.52, 2.88 and 3.45 folds, respectively, while TP was downexpressed by 0.54 fold. However, no change was observed in the methylation status of genes with FP therapy. Pairwise comparisons revealed significant difference in the expression and the methylation status of genes according to the clinicopathological characters of CRC patients either at baseline or after FP therapy. The overexpression of DPD and COX2 genes were indicators for a poor EFS of CRC patients. Also, the high level of COX2 expression was found to be significantly correlated with the hazard of progression (HR = 1.73, 95% CI = 1.02-3.03). Conclusion: The promoter methylation of FP metabolizing and COX2 genes has significant impact on the expression and the treatment outcome of CRC patients.

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Analysis of SEPT9 Gene Promoter Methylation Status in Esophageal Squamous Cell Carcinoma

Journal of Shahid Sadoughi University of Medical Sciences ◽

10.18502/ssu.v27i10.2405 ◽

2020 ◽

Author(s):

Aida Mirza Aghasi ◽

Saied Ghorbian

Keyword(s):

Squamous Cell Carcinoma ◽

Esophageal Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Squamous Cell ◽

Promoter Methylation ◽

Methylation Status ◽

Gene Promoter ◽

Sodium Metabisulfite ◽

Significant Difference ◽

Cancer Tissues

Introduction: The changes in the level of SEPT9 gene promoter methylation can contribute to the formation of esophageal squamous cell carcinoma. The aim of this study was to evaluate the level of changes in the level of SEPT9 gene promoter methylation in the esophageal squamous cell carcinoma. Methods: In the present case-control study, we collected 75 paraffin blocks of esophageal cancer tissues and 75 paraffin blocks healthy tissues, which were referred to the Noor-E-Nejat and Tabriz International Hospitals during 2013-2017. After DNA extraction and treatment with sodium metabisulfite, the changes of SEPT9 gene promoter methylation assessed using high resolution melting (HRM) technique. The data were analyzed by SPSS 22 and Chi-square test. Results: Our findings did not show a statistically significant difference between the changes of SEPT9 gene promoter methylation in cancer tissues compared to the healthy tissues (P=0.106). Conclusion: This study shows that SEPT9 gene promoter methylation cannot contribute to the esophageal squamous cell carcinoma cancerogenesis.

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Predictive value of the serum RASSF10 promoter methylation status in gastric cancer

Journal of International Medical Research ◽

10.1177/0300060519848924 ◽

2019 ◽

Vol 47 (7) ◽

pp. 2890-2900 ◽

Cited By ~ 1

Author(s):

Yilin Hu ◽

Peng Ma ◽

Ying Feng ◽

Peng Li ◽

Hua Wang ◽

...

Keyword(s):

Gastric Cancer ◽

Promoter Methylation ◽

Cox Regression ◽

Methylation Status ◽

Disease Free Survival ◽

Prognostic Indicator ◽

Chronic Atrophic Gastritis ◽

Survival Times ◽

Cox Regression Analysis ◽

Specific Pcr

Background This study aimed to investigate whether the detection of methylation in the promoter of the Ras association domain family 10 gene ( RASSF10) in the serum of patients with gastric cancer (GC) by methylation-specific PCR (MSP) can be used as a diagnostic and prognostic indicator of GC. Methods We used MSP to examine RASSF10 methylation levels in the serum and/or tumor samples from 100 GC patients, 50 patients with chronic atrophic gastritis (CAG), and 45 healthy controls (HC). We also analyzed clinicopathological and follow-up data. Results Our results showed that the rate of serum RASFF10 promoter methylation among patients with GC (49/100) was higher than in those with CAG (1/50) or HC (0/45). Moreover, the RASSF10 methylation status was consistent between serum and tumor tissues. GC patients with serum RASSF10 promoter methylation had significantly shorter overall survival and disease-free survival times than GC patients without serum RASSF10 promoter methylation. Multivariable Cox regression analysis showed that serum RASSF10 promoter methylation and lymph node metastasis both correlated with reduced survival in GC patients. Conclusions Detection of the serum RASSF10 methylation status by MSP is feasible as a diagnostic and prognostic indicator of GC.

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The Diagnosis Value of Promoter Methylation of UCHL1 in the Serum for Progression of Gastric Cancer

BioMed Research International ◽

10.1155/2015/741030 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 5

Author(s):

Gongping Wang ◽

Wei Zhang ◽

Bo Zhou ◽

Canhui Jin ◽

Zengfang Wang ◽

...

Keyword(s):

Gastric Cancer ◽

Promoter Methylation ◽

Poor Prognosis ◽

Methylation Status ◽

Tumor Stage ◽

Chronic Atrophic Gastritis ◽

Dependent Manner ◽

Specific Pcr ◽

And Control

Background.Aberrant promoter methylation has been considered as a potential molecular marker for gastric cancer (GC). However, the role of methylation of FLNC, THBS1, and UCHL1 in the development and progression of GC has not been explored.Methods.The promoter methylation status of UCHL1, FLNC, THBS1, and DLEC1 was assessed by quantitative methylation-specific PCR (QMSP) in the serum of 82 GC patients, 46 chronic atrophic gastritis (CAG) subjects, and 40 healthy controls.Results.All four genes had significantly higher methylation levels in GC patients than in CAG and control subjects. However, only UCHL1 methylation was significantly correlated with the tumor stage and lymph node metastasis. While THBS1 methylation was altered in an age-dependent manner, FLNC methylation was correlated with differentiation andHelicobacter pyloriinfection. DLEC1 methylation was only associated with tumor size. Moreover, methylated UCHL1 with or without THBS1 in the serum was found to be significantly associated with a poor prognosis.Conclusion.The promoter methylation degree of FLNC, THBS1, UCHL1, and DLEC1 in serum could tell the existence of GC and only UCHL1 in the serum was also associated with poor prognosis of GC.

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Methylation Status of miR-200b Promoter in Colorectal Polyp and Adenocarcinoma Tissues

10.32592/ircmj.2021.23.4.80 ◽

2021 ◽

Keyword(s):

Methylation Status ◽

Colorectal Polyp ◽

Cross Sectional Study ◽

Adenomatous Polyps ◽

Aberrant Methylation ◽

Hyperplastic Polyps ◽

Molecular Feature ◽

Cross Sectional ◽

Normal Tissues ◽

Specific Pcr

Background: Aberrant DNA methylation is a common molecular feature in colorectal cancer (CRC). Hypermethylation of miR-200b promoter, as an epigenetic factor, is involved in CRC tumorigenesis. The methylation status of miR-200b has been examined in CRC and adjacent normal tissues. Objectives: This study aimed to investigate miR-200b methylation in a series of colorectal adenomatous polyps, hyperplastic polyps, and adenocarcinoma tissues as precursors of CRC in the Iranian population for the first time. Materials and Methods: In this cross-sectional study (2017-2018), the methylation status of the miR-200b promoter was investigated using methylation-specific PCR in 131 fresh samples, including 30 adenocarcinoma specimens, 17 tumor-adjacent normal tissues, 78 primary lesions (55 adenomatous polyps and 23 hyperplastic polyps) and 6 healthy individuals. Results: Methylation of miR-200b was detected in adenocarcinoma samples (86%) and adenomatous polyps (85%); however, most of the hyperplastic polyps were unmethylated (69.6%). Neither control individuals nor tumor-adjacent normal tissues exhibited methylation in the miR-200b promoter. Aberrant methylation of miR-200b was significantly more common in tumor tissues and adenomatous polyps than in hyperplastic polyps (P<0.0001) and tumor-adjacent normal samples (P<0.0001). Conclusion: Methylation status of the miR-200b promoter was significantly altered during CRC development and may be identified as an attractive biomarker for the early detection of the disease.

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Promoter Methylation and mRNA Expression of Response Gene to Complement 32 in Breast Carcinoma

Journal of Cancer Epidemiology ◽

10.1155/2016/7680523 ◽

2016 ◽

Vol 2016 ◽

pp. 1-6 ◽

Cited By ~ 5

Author(s):

Ebrahim Eskandari-Nasab ◽

Mohammad Hashemi ◽

Firoozeh Rafighdoost

Keyword(s):

Mrna Expression ◽

Promoter Methylation ◽

Methylation Status ◽

Methylation Pattern ◽

Response Gene ◽

Expression Levels ◽

Promoter Methylation Status ◽

Normal Tissues ◽

A Cell ◽

Methylation Patterns

Background. Response gene to complement 32 (RGC32), induced by activation of complements, has been characterized as a cell cycle regulator; however, its role in carcinogenesis is still controversial. In the present study we comparedRGC32promoter methylation patterns and mRNA expression in breast cancerous tissues and adjacent normal tissues.Materials and Methods. Sixty-three breast cancer tissues and 63 adjacent nonneoplastic tissues were included in our study.Design. Nested methylation-specific polymerase chain reaction (Nested-MSP) and quantitative PCR (qPCR) were used to determineRGC32promoter methylation status and its mRNA expression levels, respectively.Results. RGC32 methylation pattern was not different between breast cancerous tissue and adjacent nonneoplastic tissue (OR = 2.30, 95% CI = 0.95–5.54). However, qPCR analysis displayed higher levels ofRGC32mRNA in breast cancerous tissues than in noncancerous tissues (1.073 versus 0.959;P=0.001), irrespective of the promoter methylation status. The expression levels and promoter methylation ofRGC32were not correlated with any of patients’ clinical characteristics (P>0.05).Conclusion. Our findings confirmed upregulation of RGC32 in breast cancerous tumors, but it was not associated with promoter methylation patterns.

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MS-FLAG, a Novel Real-Time Signal Generation Method for Methylation-Specific PCR

Clinical Chemistry ◽

10.1373/clinchem.2007.094011 ◽

2007 ◽

Vol 53 (12) ◽

pp. 2119-2127 ◽

Cited By ~ 27

Author(s):

Cinzia Bonanno ◽

Erlet Shehi ◽

Daniel Adlerstein ◽

G Mike Makrigiorgos

Keyword(s):

Real Time ◽

Promoter Methylation ◽

Real Time Pcr ◽

Bisulfite Sequencing ◽

Methylation Status ◽

Gene Promoter ◽

Signal Generation ◽

Time Signal ◽

Specific Pcr ◽

Generation Technology

Abstract Background: Aberrant promoter methylation is a major mechanism for silencing tumor suppressor genes in cancer. Detection of hypermethylation is used as a molecular marker for early cancer diagnosis, as a prognostic index, or to define therapeutic targets for reversion of aberrant methylation. We report on a novel signal generation technology for real-time PCR to detect gene promoter methylation. Methods: FLAG (fluorescent amplicon generation) is a homogeneous signal generation technology based on the exceptionally thermostable endonuclease PspGI. FLAG provides real-time signal generation during PCR by PspGI-mediated cleavage of quenched fluorophores at the 5′ end of double-stranded PCR products. Methylation-specific PCR (MSP) applied on bisulfite-treated DNA was adapted to a real-time format (methylation-specific FLAG; MS-FLAG) for quantifying methylation in the promoter of CDKN2A (p16), GATA5, and RASSF1. We validated MS-FLAG on plasmids and genomic DNA with known methylation status and applied it to detection of methylation in a limited number of clinical samples. We also conducted bisulfite sequencing on these samples. Results: Real-time PCR results obtained via MS-FLAG agreed with results obtained via conventional, gel-based MSP. The new technology showed high specificity, sensitivity (2–3 plasmid copies), and selectivity (0.01% of methylated DNA) on control samples. It enabled correct prediction of the methylation status of all 3 gene promoters in 21 lung adenocarcinoma samples, as confirmed by bisulfite sequencing. We also developed a multiplex MS-FLAG assay for GATA5 and RASSF1 promoters. Conclusion: MS-FLAG provides a new, quantitative, high-throughput method for detecting gene promoter methylation and is a convenient alternative to agarose gel-based MSP for screening methylation. In addition to methylation, FLAG-based real-time signal generation may have broad applications in DNA diagnostics.

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Epigenetic Inactivation of Protocadherin 10 by Methylation in Colorectal Cancer

ACTA MEDICA IRANICA ◽

10.18502/acta.v57i8.2410 ◽

2020 ◽

Author(s):

Mohammad Amin Kerachian ◽

Sahar Tavakolian ◽

Matineh Barati Bagherabad ◽

Dor Mohammad Kordi-Tamandani ◽

Mohammad Reza Abbaszadegan

Keyword(s):

Colorectal Cancer ◽

Tumor Suppressor ◽

Promoter Methylation ◽

Methylation Status ◽

Cpg Islands ◽

High Specificity ◽

Suppressor Gene ◽

Potential Candidate ◽

Critical Function ◽

Normal Tissues

Aberrant promoter methylation of CpG islands of tumor-suppressor genes has been recognized as one of the important tumor markers for cancer detection. The aim of this study was to investigate the promoter methylation status of protocadherin 10 (PCDH10), a tumor suppressor gene, in Iranian colorectal cancer (CRC) patients. Cancerous and the adjacent normal tissues obtained from 38 CRC patients were used to assess the methylation status of PCDH10 with Methylation Specific PCR, in addition, to study the expression level of this gene by quantitative PCR. The relationship between hypermethylation and the demographic characteristics of these patients was analyzed. The promoter methylation level of PCDH10 was statistically different between tumoral and normal tissues in CRC patients. Twenty-seven out of 38 patients showed hypermethylation with a sensitivity of 73% and a specificity of 97%. PCDH10 expression decreased in 15 cases (46%) as 16 cases (50 %) showed overexpression and 1 case (4%) had no changes. Not a significant association was reported between PCDH10 hypermethylation and the clinicopathological characteristics (P>0.05). Our results indicated that PCDH10 methylation has a critical function in CRC, with a nearly elevated sensitivity and a high specificity in the Iranian population, qualify it as a potential candidate biomarker. © 2019 Tehran University of Medical Sciences. All rights reserved. Acta Med Iran 2019;57(8):472-477.

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