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DNA修复基因MGMT在乳腺浸润性导管癌中的蛋白表达、甲基化及其与临床病理参数的相关性

Protein expression and methylation of MGMT, a DNA repair gene and their correlation with clinicopathological parameters in invasive ductal carcinoma of the breast.

作者信息

Asiaf Asia, Ahmad Shiekh Tanveer, Malik Ajaz Ahmad, Aziz Shiekh Aejaz, Rasool Zubaida, Masood Akbar, Zargar Mohammad Afzal

机构信息

Department of Biochemistry, Faculty of Biological Science, University of Kashmir, Srinagar, J&K, India, 190006,

出版信息

Tumour Biol. 2015 Aug;36(8):6485-96. doi: 10.1007/s13277-015-3339-9. Epub 2015 Mar 29.

Abstract

Epigenetic mechanisms such as DNA methylation are being increasingly recognized to play an important role in cancer and may serve as a cancer biomarker. The aim of this study was to evaluate the promoter methylation status of MGMT (O6-methylguanine-DNA methyltransferase) and a possible correlation with the expression of MGMT and standard clinicopathological parameters in invasive ductal breast carcinoma patients (IDC) of Kashmir. Methylation-specific PCR was carried out to investigate the promoter methylation status of MGMT in breast tumors paired with the corresponding normal tissue samples from 128 breast cancer patients. The effect of promoter methylation on protein expression in the primary breast cancer and adjacent normal tissues was evaluated by immunohistochemistry (n = 128) and western blotting (n = 30). The frequency of tumor hypermethylation was 39.8 % and a significant difference in methylation frequency among breast tumors were found (p < 0.001) when compared with the corresponding normal tissue. Immunohistochemical analysis showed no detectable expression of MGMT in 68/128 (53.1 %) tumors. MGMT promoter methylation mediated gene silencing was associated with loss of its protein expression (rs = -0.285, p = 0.001, OR = 3.38, 95 % CI = 1.59-7.17). A significant correlation was seen between loss of MGMT and lymph node involvement (p = 0.030), tumor grade (p < 0.0001), loss of estrogen receptors (ER; p = 0.021) and progesterone receptors (PR) (p = 0.016). Also, MGMT methylation was found to be associated with tumor grade (p = 0.011), tumor stage (p = 0.009), and loss of ER (p = 0.003) and PR receptors (p = 0.009). To our knowledge, our findings, for the first time, in Kashmiri population, indicate that MGMT is aberrantly methylated in breast cancer and promoter hypermethylation could be attributed to silencing of MGMT gene expression in breast cancer. Our data suggests that MGMT promoter hypermethylation could have a potential function as molecular biomarker of breast oncogenesis. Also, based on their predictive value of response to therapy, the immunohistochemical evaluation and interpretation of MGMT may also help in future to establish therapeutic strategies for patients with breast cancer.

摘要

诸如DNA甲基化等表观遗传机制在癌症中发挥的重要作用正日益得到认可,并且可能作为一种癌症生物标志物。本研究的目的是评估克什米尔浸润性导管癌患者(IDC)中MGMT(O6-甲基鸟嘌呤-DNA甲基转移酶)的启动子甲基化状态以及与MGMT表达和标准临床病理参数之间的可能相关性。采用甲基化特异性PCR研究了128例乳腺癌患者的乳腺肿瘤及其相应正常组织样本中MGMT的启动子甲基化状态。通过免疫组织化学(n = 128)和蛋白质印迹法(n = 30)评估启动子甲基化对原发性乳腺癌和相邻正常组织中蛋白质表达的影响。肿瘤高甲基化频率为39.8%,与相应正常组织相比,乳腺肿瘤之间的甲基化频率存在显著差异(p < 0.001)。免疫组织化学分析显示,68/128(53.1%)的肿瘤中未检测到MGMT表达。MGMT启动子甲基化介导的基因沉默与其蛋白质表达缺失相关(rs = -0.285,p = 0.001,OR = 3.38,95%CI = 1.59 - 7.17)。MGMT缺失与淋巴结受累(p = 0.030)、肿瘤分级(p < 0.0001)、雌激素受体(ER;p = 0.021)和孕激素受体(PR)缺失(p = 0.016)之间存在显著相关性。此外,发现MGMT甲基化与肿瘤分级(p = 0.011)、肿瘤分期(p = 0.009)以及ER(p = 0.003)和PR受体缺失(p = 0.009)相关。据我们所知,我们的研究结果首次表明,在克什米尔人群中,MGMT在乳腺癌中存在异常甲基化,启动子高甲基化可能导致乳腺癌中MGMT基因表达沉默。我们的数据表明,MGMT启动子高甲基化可能具有作为乳腺肿瘤发生分子生物标志物的潜在功能。此外,基于其对治疗反应的预测价值,MGMT的免疫组织化学评估和解读未来可能也有助于为乳腺癌患者制定治疗策略。

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