Methylation Analysis of DNA Mismatch Repair Genes Using DNA Derived from the Peripheral Blood of Patients with Endometrial Cancer: Epimutation in Endometrial Carcinogenesis

Takeda, Takashi; Banno, Kouji; Yanokura, Megumi; Adachi, Manabu; Iijima, Moito; Kunitomi, Haruko; Nakamura, Kenzo; Iida, Miho; Nogami, Yuya; Umene, Kiyoko; Masuda, Kenta; Kobayashi, Yusuke; Yamagami, Wataru; Hirasawa, Akira; Tominaga, Eiichiro; Susumu, Nobuyuki; Aoki, Daisuke

doi:10.3390/genes7100086

Cited by 9 publications

(9 citation statements)

References 40 publications

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“…Hypermethylation could be the 'second hit', exerting an important impact on the regulation of MMR genes expression. 30 MLH1 promoter methylation is the primary cause of reduced MMR capacity in multiple human cancers due to the low expression of MLH1 protein. 31 But in Lynch syndrome-associated tumours (positive for germline MMR genes mutations), MLH1 methylation is a rare event that sensitive and specific detection of it could be important to distinguish whether the tumour samples are from Lynch syndrome or other cancers.…”

Section: Discussionmentioning

confidence: 99%

“…The high detection rate of germline MMR genes mutations in EMPD from our previous study provided a potential link between EMPD and Lynch syndrome, with both of these diseases sharing the similar molecular mechanisms in the pathological process. Hypermethylation could be the ‘second hit’, exerting an important impact on the regulation of MMR genes expression . MLH1 promoter methylation is the primary cause of reduced MMR capacity in multiple human cancers due to the low expression of MLH1 protein .…”

Section: Discussionmentioning

confidence: 99%

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Methylation and expression analysis of mismatch repair genes in extramammary Paget's disease

Kang

Zhu

Zhang

et al. 2019

Acad Dermatol Venereol

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Background Extramammary Paget's disease (EMPD) is a rare skin cancer with relative high frequencies of germline and somatic mismatch repair (MMR) genes mutations. However, the methylation and expression of these genes have not been validated in EMPD. Objective This study aims to confirm the methylation and expression of MMR genes in EMPD.Methods Immunohistochemical (IHC) staining detection and Methylation-specific PCR (MSP) were used to analyse MLH1, MSH2, MSH6 and PMS2 proteins' expression and promoters' methylation in 57 EMMD samples, and pyrosequence was used to find highly methylated CpG sites in MSH2 promoter.Results Immunohistochemical detection displayed reduced expression of MSH2 in 38.6% EMPD cases but normal expression of MLH1, MSH6 and PMS2 in all tumour tissues. Hypermethylation also was found in the promoter of MSH2 but not in other MMR genes. Pyrosequencing of MSH2 promoter showed CpG6 (-87) and CpG3 (-98) were the most common two methylated CpG dinucleotides. There is a significant correlation between reduced MSH2 expression and MSH2 methylation.Conclusion Reduced MSH2 expression and hypermethylation in this gene promoter were common genetic changes in EMPD, which expands our understanding of the role of MMR function in this skin cancer.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Methylation and expression analysis of mismatch repair genes in extramammary Paget's disease

Kang

Zhu

Zhang

et al. 2019

Acad Dermatol Venereol

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“…Decreased DNA methylation occurs early in carcinogenesis, and promoter hypermethylation leads to gene silencing and loss of gene expression. Therefore, carcinogenesis can be induced when the tumour suppressor gene or a critical gene involved in the cell cycle or in DNA repair is affected ( 53 , 54 ).…”

Section: Discussionmentioning

confidence: 99%

Methylation status of KLF4 and HS3ST2 genes as predictors of endometrial cancer and hyperplastic endometrial lesions

et al. 2018

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Endometrial carcinoma is one of the most common tumours in developed countries. In addition to the active role of genetic factors, epigenetic changes also have an important effect. The present study analysed the methylation status of kruppel like factor 4 (KLF4) and heparan sulfate-glucosamine 3-sulfotransferase 2 (HS3ST2) genes in three endometrial tissue types for carcinoma prediction. The sample comprised 91 women with histologically-confirmed endometrial carcinoma (64.16±9.64 years old), 36 women with hyperplasia (53.39±9.64 years old) and 45 with no signs or symptoms of malignancy (48.53±11.11 years old). The CpG dinucleotide methylation levels were examined by quantitative pyrosequencing, and the discrimination accuracy of the model was calculated using the Random Forest classification algorithm of the area under the ROC curve (AUC). The mean values of KLF4 and HS3ST2 methylation indices were 23.83±11.39 and 8.52±2.57 in the control samples; 30.40±8.52 and 33.76±20.66 in hyperplasia and 34.72±10.79 and 34.49±18.39 in the cancerous tissues. Multinomial logistic regression indicated that the HS3ST2 CpG1 methylation status is a predictor of hyperplasia (P<0.05) and that the KLF4 CpG2 dinucleotide can predict carcinoma formation (P<0.001). The AUC value of 0.95 indicates high discrimination accuracy of the CpG nucleotides methylation status model between the controls and the two other diagnoses. The results of the present study establish the likelihood that aberrations in KLF4 and HS3ST2 gene methylation levels are important in the development of endometrial hyperplasia and carcinoma, with hyperplasia an intermediate step between healthy and tumour tissues.

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“…Immunohistochemical analysis was performed as previously described [ 16 ]. Tissue was fixed overnight with 4% paraformaldehyde, embedded in paraffin, and prepared as sections of 4 μm.…”

Section: Methodsmentioning

confidence: 99%

“…The methylation status of CpG islands in the MLH1 , MSH2 and MSH6 promoters was analyzed by MSP. This method detects methylation of bases by a bisulfite reaction and subsequent detection by polymerase chain reaction (PCR) with primers specific to bisulfite-converted methylated and unmethylated sequences [ 16 17 ]. MSP primer sets for MLH1, MSH2, and MSH6 are summarized in Supplementary Table 1 .…”

Section: Methodsmentioning

confidence: 99%

Screening for Lynch syndrome using risk assessment criteria in patients with ovarian cancer

et al. 2018

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ObjectiveLynch syndrome is a cancer predisposition syndrome caused by germline mutation of DNA mismatch repair (MMR) genes. Lynch syndrome only causes about 0.4% of cases of ovarian cancer, which suggests that universal screening may not be cost-efficient. However, the frequency of Lynch syndrome in ovarian cancer is unclear in the Asian population. The goal of the study was to investigate a screening strategy using family history.MethodsThe subjects were 129 patients with ovarian cancer. Clinical and family history were collected using a self-administered questionnaire, and Society of Gynecologic Oncology (SGO) criteria 2007 and PREMM5 were used for risk assessment. Microsatellite instability, immunohistochemistry, and methylation of MMR genes were analyzed.ResultsOf the 129 cases, 25 (19.4%) met the SGO criteria, and 4 of these 25 had MSI-high and MMR deficiency. Two cases had loss of MSH2 and MSH6, indicating MSH2 mutation, and the other two had loss of MLH1 and PMS2, including one without MLH1 methylation indicating MLH1 mutation. These results show that screening using family history can detect Lynch syndrome in 12.0% (3/25) of ovarian cancer cases. The 3 cases were positive for PREMM5, but negative for Amsterdam II criteria and revised Bethesda guidelines. Genetic testing in one case with MSH2 and MSH6 deficiency confirmed the diagnosis of Lynch syndrome with MSH2 mutation.ConclusionThis is the first study of screening for Lynch syndrome in ovarian cancer using clinical and family history in an Asian population. This approach may be effective for diagnosis in these patients.

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Methylation Analysis of DNA Mismatch Repair Genes Using DNA Derived from the Peripheral Blood of Patients with Endometrial Cancer: Epimutation in Endometrial Carcinogenesis

Cited by 9 publications

References 40 publications

Methylation and expression analysis of mismatch repair genes in extramammary Paget's disease

Methylation and expression analysis of mismatch repair genes in extramammary Paget's disease

Methylation status of KLF4 and HS3ST2 genes as predictors of endometrial cancer and hyperplastic endometrial lesions

Screening for Lynch syndrome using risk assessment criteria in patients with ovarian cancer

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