Correlation of WIF-1 expression and clinicopathologic significance in breast cancer with aberrant DNA methylation
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摘要:
目的 研究WIF-1（Wnt inhibitory factor-1）mRNA在乳腺癌组织中的表达及其启动子区域甲基化情况，进一步探讨WIF-1基因甲基化与乳腺癌临床病理特征的关系。 方法 收集2009年9月1日至2009年12月30日青岛大学附属医院乳腺外科手术切除新鲜组织标本69例，其中良性病变组织9例，乳腺癌及癌旁组织各30例，应用RT-PCR及甲基化特异性PCR（methylation specific PCR，MSP）检测乳腺癌组织、相应癌旁组织和乳腺良性病变组织中WIF-1mRNA表达及其启动子甲基化情况。 结果 癌组织中WIF-1基因表达率明显低于相应癌旁组织及乳腺良性病变组织，具有显著性差异（χ2=41.786，P < 0.05）；与其他两组相比甲基化率在癌组织中明显升高（矫正χ2=16.484，P < 0.05）；WIF-1基因表达下降与其异常甲基化存在明显关联（P=0.023）；WIF-1异常甲基化与乳腺癌发病年龄、肿瘤分级、组织分型和淋巴结转移无相关性（P>0.05）。 结论 异常甲基化可能是乳腺癌WIF-1基因表达下降的重要原因，是乳腺癌发生、发展的重要机制。 Abstract:Objective To investigate the mRNA expression of the WIF-1 gene and the methylation of its promoter in breast cancer, and to determine the correlation between the epigenetic aberrant WIF-1 DNA methylation and the clinicopathological significance of WIF-1 in breast cancer. Methods RT-PCR and sensitive methylation-specific-PCR (MSP) were used to detect WIF-1 mRNA expression and the methylation of the WIF-1 promoter in 30 breast cancer samples as well as in tumor-adjacent tissue samples and 9 benign breast tissues. Results The WIF-1 mRNA expression in 30 breast cancer samples significantly decreased compared with those of the other two groups. In addition, WIF-1 methylation was more frequent in breast-tumor tissues compared with those in tumor-free tissues. Meanwhile, WIF-1 mRNA expression in breast cancer tissues involved the abnormal methylation of its promoter. Clinicopathological correlation analysis showed that the abnormal methylation of the WIF-1 gene promoter was not associated with age, TNM stage, histotype, or lymph node metastasis. Conclusion WIF-1 mRNA expression loss due to abnormal methylation may be a crucial factor in breast cancer development and can thus be used in the prognosis and progression of the disease. -
Key words:
- WIF-1 /
- gene expression /
- breast cancer /
- methylation /
- TNM stage /
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图 1 乳腺癌组织、乳腺癌癌旁组织及乳腺良性病变组织中的WIF-1mRNA表达
Figure 1. mRNA analysis of WIF-1 in breast cancer tissue, adjacent non-cancerous tissue, and hyperplasic breast tissue
M: DNA marker; 1, 3, 5, and 7:breast cancer tissue; 2, 4, 6, and 8: Tumor-adjacent normal tissue; 9:Benign-breast disease tissue; H2O: Blank control
图 2 乳腺癌组织、乳腺癌癌旁组织及乳腺良性病变组织中的WIF-1甲基化状况
Figure 2. Analysis of WIF-1 methylation in breast cancer tissue, adjacent non-cancerous tissue, and hyperplasic breast tissue
P: Positive control; T: Breast cancer tissue; N: Tumor-adjacent normal tissue; B: Benign-breast disease tissue; H2O: Blank control; U: Un⁃ methylated DNA; M: Methylated DNA
图 3 BGS检测WIF-1基因启动子甲基化状态测序图
Figure 3. BGS analysis of the methylation of the WIF-1 promoter region
表 1 PCR检测引物序列
Table 1. Primer sequences and PCR conditions
3 BGS检测WIF-1基因启动子CpG位点甲基化状态结果
3. BGS analysis of the methylation of the WIF-1 promoter region
表 3 乳腺癌组织WIF-1甲基化与临床特征的关系
Table 3. Relationship between WIF-1 methylation in breast cancer tissue and clinical features
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