Methylation Patterns of Cancer-Associated Genes in Breast Cancer

Sung-Bae Jee; Woo-Chan Park; Kee-Whan Kim; Ji-Il Kim; Chang-Hyeok Ahn; Keun-Woo Lim; Se-Jung Oh; Byung-Joo Song; Sang-Seol Jung; Jeong-Soo Kim

doi:10.4048/jbc.2007.10.1.51

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Jee, Park, Kim, Kim, Ahn, Lim, Oh, Song, Jung, and Kim: Methylation Patterns of Cancer-Associated Genes in Breast Cancer

Original Article

Journal of Breast Cancer

Journal of Breast Cancer 2007; 10(1): 51-58.

Published online: 31 March 2007

DOI: https://doi.org/10.4048/jbc.2007.10.1.51

Methylation Patterns of Cancer-Associated Genes in Breast Cancer

Sung-Bae Jee, Woo-Chan Park, Kee-Whan Kim, Ji-Il Kim, Chang-Hyeok Ahn, Keun-Woo Lim, Se-Jung Oh, Byung-Joo Song, Sang-Seol Jung, Jeong-Soo Kim

Department of Surgery, The Catholic University of Korea, College of Medicine, Seoul, Korea.

drbreast@catholic.ac.kr

Received 11 December 2006 Accepted 23 February 2007

Abstract

Purpose

To investigate the methylation status of cancer-associated genes in breast cancer to assess its use in the diagnosis of breast cancer and the relationship with distinctive clinical and pathological features.

Methods

A total of 29 benign tumors and their adjacent normal tissues as well as 67 malignant tumors and adjacent normal samples, from women undergoing surgery for primary invasive breast carcinoma at Uijongbu St. Mary's Hospital, between March 2003 and March 2005, were used. Eleven candidate genes were chosen; P14, P16, DAPK, MGMT, h-MLH, E-cadherin, RASSF1α, Twist, RARβ, HIN-1, and Cyclin D. DNA was extracted from fresh tissues, and methylation specific PCR performed.

Results

The number of methylated genes was increased in the malignant tissues compared to the benign tumors and adjacent normal tissues. 7 genes; P14, P16, MGMT, RASSF1α, Twist, RARβ, and Cyclin D, were more frequently methylated in malignant than benign tumors, with the differences in the p14, p16, and RARβ, genes were statistically significant (p<0.05). In benign tumors, the p16 and HIN-1 genes were the most infrequently (6.9%) and frequently methylated (82.8%), respectively. In malignant tumors, the h-MLH and RASSF1α genes were most infrequently and frequently methylated genes, respectively. The subgroup showing methylation of the DAPK gene had a higher nuclear grade and greater progesterone receptor negativity. The group in which the RASSF1α gene was methylated, had greater estrogen receptor (ER) and progesterone receptor (PgR) positivities. The Twist gene was frequently methylated in the subgroup showing higher nuclear and histologic grades. The group with HIN-1 and cyclin D methylation had a tendency to show greater ER positivity.

Conclusion

The subgroups showing methylated DAPK and Twist should be more intensely treated and followed up more carefully than those with RASSF1α, HIN-1 and Cyclin D methylation. Gene methylation may be linked to various pathological features of breast cancer; however, this will require confirmation from larger studies.

Keywords: Breast cancer, Tumor suppressor gene, Methylation

Figures and Tables

Fig 1

Number of methylated genes, benign tumor vs. normal tissue.

Fig 2

Number of methylated genes, malignant tumor vs. normal tissue.

Fig 3

The correlation of numbers of methylated genes between normal tissue vs. malignant tumor.

Fig 4

The correlation of numbers of methylated genes between normal tissue vs. benign tumor.

Table 1

Genes investigated in this study

Abb.=abbreviation.

Table 2

Characteristics of breast cancer patients

BCS=breast conserving surgery; MRM=modified radical mastectomy.

Table 3

MS-PCR primers of specific genes analyzed in this study

Table 4

Comparision of numbers of hypermethylation in 11 genes

^*: Paired T-test; ^†: Independent T-test.

Table 5

The comparison of methylation status of each gene, benign tumor vs. malignant tumor

^*: Chi-square Test; ^†: Fischer exact Test.

Table 6

The comparison of methylation status of each gene, normal tissue vs. malignant tumor

^*: Chi-square Test.

Table 7

Logistic regression of hypermethylation status, benign tumor vs. malignant tumor

Exp(B)=odds ratio.

Table 8

Logistic regression of hypermethylation status, normal tissue vs. malignant tumor

Exp(B)=odds ratio.

Table 9

Association between gene promoter methylation and clinicopathological features

^*: Chi-square test; ^†: Linear by linear test; ^‡: Fischer exact test.

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