Cancer Res Treat.  2005 Aug;37(4):233-240.

Gene Promoter Hypermethylation in Tumors and Plasma of Breast Cancer Patients

• Bae YK
• Shim YR
• Choi JH
• Kim MJ
• Gabrielson E
• Lee SJ
• Hwang TY
• Shin SO

Abstract

PURPOSE
To measure the hypermethylation of four genes in primary tumors and paired plasma samples to determine the feasibility of gene promoter hypermethylation markers for detecting breast cancer in the plasma. MATERIALS AND METHODS: DNA was extracted from the tumor tissues and peripheral blood plasma of 34 patients with invasive breast cancer, and the samples examined for aberrant hypermethylation in cyclin D2, retinoic acid receptor beta (RARbeta), twist and high in normal-1 (HIN-1) genes using methylation-specific PCR (MSP), and the results correlated with the clinicopathological parameters. RESULTS: Promoter hypermethylation was detected at high frequency in the primary tumors for cyclin D2 (53%), RARbeta (56%), twist (41%) and HIN-1 (77%). Thirty-three of the 34 (97%) primary tumors displayed promoter hypermethylation in at least one of the genes examined. The corresponding plasma samples showed hyperme thylation of the same genes, although at lower frequencies (6% for cyclin D2, 16% for RARbeta, 36% for twist, and 54% for HIN-1). Overall, 22 of the 33 (67%) primary tumors with hypermethylation of at least one of the four genes also had abnormally hypermethylated DNA in their matched plasma samples. No significant relationship was recognized between any of the clinical or pathological parameters (tumor size, axillary lymph node metastasis, stage, or Ki-67 labeling index) with the frequency of hypermethylated DNA in the primary tumor or plasma. CONCLUSION: The detection of aberrant promoter hypermethylation of cancer-related genes in the plasma may be a useful tool for the detection of breast cancer.

Keyword

Methylation; Plasma; Breast neoplasms

MeSH Terms

Breast Neoplasms*
Breast*
Cyclin D2
DNA
Humans
Lymph Nodes
Methylation
Neoplasm Metastasis
Plasma*
Polymerase Chain Reaction
Receptors, Retinoic Acid
Cyclin D2
DNA
Receptors, Retinoic Acid

Figure

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  Fig. 1 Representative MSP results in breast cancer tissues, their paired plasma samples and normal controls. Lanes labeled "M" represent reactions using primers specific for bisulfite-treated DNA products with methylated CpG sites, and lanes labeled "U" represent reactions using primers specific for bisulfite-treated DNA products with unmethylated CpG sites. Some normal controls showed no amplified signals for methylated or unmethylated reactions. In those cases, the MSP was repeated and the absence of a methylated signal, in contrast to the presence of an unmethylated signal, confirmed.

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  Fig. 2 Clinicopathological data and MSP results of breast cancer patients. Black and white boxes indicate methylated and unmethylated samples, respectively. Criteria for high Ki-67 is nuclear staining in more than 30% of tumor cells.

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  Fig. 3 Distribution of patients according to the number of methylated genes in the primary breast tumor and the plasma. The numerals in the box indicate the number of methylated genes.

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