Optimization of High Resolution Melting (HRM) Method for Genotyping and Estimating Frequency of SNP rs4284505 in the Vietnamese Breast Cancer

Background and Objective. Breast cancer is the leading cause of death from cancer among women worldwide. The aberrant promoter methylation of tumor suppressor genes is a typical epigenetic alteration for breast cancer and can be detected in early carcinogenesis. High-throughput and cost-effective methods are needed for the early and sensitive detection of epigenetic changes in clinical material. The main purpose of our study was to optimize a high-resolution melting (HRM) assay for the reliable and quantitative assessment of RASSF1 gene methylation, which is considered one of the earliest epigenetic alterations in breast cancer. Material and Methods. A total of 76 breast carcinomas and 10 noncancerous breast tissues were studied by means of HRM and compared with the results obtained by means of quantitative methylation-specific polymerase chain reaction (QMSP) and methylation-specific polymerase chain reaction (MSP). Results. Both quantitative methods, HRM and QMSP, showed a similar specificity and sensitivity for the detection of RASSF1 methylation in breast cancer (about 80% and 70%, respectively). In breast cancer, the mean methylation intensity of RASSF1 was 42.5% and 48.6% according to HRM and QMSP, respectively. Both methods detected low levels of methylation (less than 5%) in noncancerous breast tissues. In comparison with quantitative methods, MSP showed a lower sensitivity (70%), but a higher specificity (80%) for the detection of RASSF1 methylation in breast cancer. Conclusions. HRM is as a simple, cost-effective method for the reliable high-throughput quantification of DNA methylation in clinical material.

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High Resolution Melting (HRM) Optimization for Genotyping the SNP Rs3746444/Mir-499 in Vietnamese Breast Cancer Patients

6th International Conference on the Development of Biomedical Engineering in Vietnam (BME6) - IFMBE Proceedings ◽

10.1007/978-981-10-4361-1_29 ◽

2017 ◽

pp. 171-176

Author(s):

Uyen Hong Thi Nguyen ◽

Thanh Ngoc Thi Nguyen ◽

Thiep Van Tran ◽

Hue Thi Nguyen

Keyword(s):

Breast Cancer ◽

High Resolution ◽

Cancer Patients ◽

High Resolution Melting ◽

Breast Cancer Patients

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P3.06 Comparison of High Resolution Melting and Restriction Fragment Length Polymorphism to Genotype Snp in Formalin Fixed Paraffin-Embedded Breast Cancer Tissue

Annals of Oncology ◽

10.1016/s0923-7534(20)31359-4 ◽

2012 ◽

Vol 23 ◽

pp. v34

Author(s):

R. Oktriani ◽

A. Wahyono ◽

S. Haryono ◽

A. Utomo ◽

T. Aryandono

Keyword(s):

Breast Cancer ◽

High Resolution ◽

High Resolution Melting ◽

Fragment Length Polymorphism ◽

Breast Cancer Tissue ◽

Cancer Tissue ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Formalin Fixed ◽

Restriction Fragment Length

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Optimizing high-resolution melting analysis for the detection of mutations of GPR30/GPER-1 in breast cancer

Gene ◽

10.1016/j.gene.2012.04.029 ◽

2012 ◽

Vol 501 (2) ◽

pp. 118-126 ◽

Cited By ~ 6

Author(s):

Masamune Aihara ◽

Shigeru Yamamoto ◽

Hiroko Nishioka ◽

Yutaro Inoue ◽

Kimikazu Hamano ◽

...

Keyword(s):

Breast Cancer ◽

High Resolution ◽

High Resolution Melting ◽

High Resolution Melting Analysis ◽

Melting Analysis ◽

Detection Of Mutations

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Developing a high resolution melting method for genotyping and predicting association of SNP rs353291 with breast cancer in the Vietnamese population

Biomedical Research and Therapy ◽

10.15419/bmrat.v4i12.387 ◽

2017 ◽

Vol 4 (12) ◽

pp. 1812 ◽

Cited By ~ 2

Author(s):

Luan Huu Huynh ◽

Phuong Thi-Kim Bui ◽

Thanh Thi-Ngoc Nguyen ◽

Hue Thi Nguyen

Keyword(s):

Breast Cancer ◽

High Resolution ◽

High Resolution Melting ◽

Risk Allele ◽

Nucleotide Polymorphisms ◽

Single Nucleotide ◽

P Values ◽

Potential Biomarker ◽

Vietnamese Population ◽

The One

Introduction: Breast cancer is the one of the most common types of cancer as well as the second leading cause of cancer death in women in the world. In recent studies, microRNAs (miRNAs) have been demonstrated to play a crucial role as a new potential biomarker in the association with breast cancer. Single Nucleotide Polymorphisms (SNPs) located on specific miRNA may result in breast cancer. Among the SNPs, SNP rs353291 has shown to be associated with breast cancer in individuals of Caucasian background. Furthermore, this SNP is observed in a high percentage of mutant alleles in the Vietnamese population. Thus, SNP rs353291 was selected as a candidate SNP for investigation in this study. The frequency of SNP rs353291 was evaluated by High Resolution Melting (HRM) method, which is a highly powerful method to detect variants in DNA sequence, especially for SNP genotyping. Methods: In this study, the association between this SNP and risk of breast cancer in the Vietnamese population was evaluated in 90 cases and 96 healthy controls via genotyping using an optimized HRM protocol. Result: The genotyping results revealed that SNP rs353291 is a polymorphism in the Vietnamese population. We have successfully identified frequencies of AA, AG and GG to be 40%, 42.2% and 17.8%, respectively. In particular, the calculated frequencies of allele G was 61.1% while risk allele A was 38.9%. The association between this SNP and breast cancer in Vietnam revealed that there is an obvious decreased risk of breast cancer among Vietnamese population when comparing G allele to A allele (G vs A: OR=0.92, 95% CI: 0.62-1.36, p= 0.677); the results also showed that heterozygote model had a reduced risk of breast cancer compared to dominant model (GA+GG vs AA: OR=0.94, 95% CI: 0.52-1.70, p=0.839). Conclusion: However, since the p-values were >0.05, our results only show a correlation rather than a significant association between SNP rs353291 and breast cancer risk in the Vietnamese population.

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