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首页> 外文期刊>European journal of medical research. >Methylated APC and GSTP1 Genes in Serum DNA Correlate with the Presence of Circulating Blood Tumor Cells and are Associated with a More Aggressive and Advanced Breast Cancer Disease.
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Methylated APC and GSTP1 Genes in Serum DNA Correlate with the Presence of Circulating Blood Tumor Cells and are Associated with a More Aggressive and Advanced Breast Cancer Disease.

机译：血清DNA中的甲基化APC和GSTP1基因与循环血液肿瘤细胞的存在相关，并且与更激进和晚期的乳腺癌疾病相关。

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Background: Tumor-related methylated DNA and circulating tumor cells (CTC) in the peripheral blood might be of prognostic importance in breast cancer. Thus, the aim of our study was to examine free methylated DNA and CTC in the blood from breast cancer patients and to correlate it with clinicopathological features known to influence prognosis. - Materials and Methods: We prospectively obtained serum samples from 85 patients with breast cancer and 22 healthy volunteers. Sera were analysed by methylation specific PCR (MethyLight PCR) for five genes: adenomatous polyposis coli (APC), ras association domain family protein 1A (RASSF1A), estrogen receptor 1 (ESR1), CDKN2A (p16) and glutathione s-transferase pi 1 (GSTP1). Beta actin (ACTB) served as control. In parallel matched peripheral blood of 63 patients was used to assay for circulating tumor cells in the peripheral blood by a modified immunomagnetic AdnaTest BreastCancerSelect with PCR detection for EPCAM, MUC1, MGB1 and SPDEF. - Results: A hypermethylation in the APC gene in 29% (25/85), in RASSF1A in 26% (22/85), in GSTP1 in 18% (14/76) and in ESR1 in 38% (32/85) of all breast cancer patients was detected. No hypermethylation of CDKN2A was found (0/25). Blood samples of patients were defined CTC positive by detecting the EPCAM 13% (8/63), MUC1 16% (10/63), MGB 9% (5/55), SPDEF 12% (7/58) and in 27% detecting one or more genes (15/55). A significant difference was seen in methylated APC DNA between cancer patients and healthy volunteers. Moreover, methylated APC, RASSF1 and CTC were significantly different in metastatic versus non-metastatic disease. In addition, the presence of methylated APC, RASSF1A and CTC correlated significantly with AJCC-staging (p = 0.001, p = 0.031 and 0.002, respectively). High incidences of methylations were found for the genes RASSF1 and ESR1 in healthy individuals (both 23% 5/22). Methylated GSTP1 was predominantly found in the serum of patients with large primaries (p = 0.023) and was highly significantly correlated with positive Her2eu status (p = 0.003). Elevated serum CA15.3 was strongly correlated with methylated APC and CTC detection (both p = 0.000). Methylated ESR1 failed to exhibit significant correlations with any of the above mentioned parameters. The presence of CTC in peripheral blood was significantly associated with methylated APC (p = 0.012) and methylated GSTP1 (p = 0.001). - Conclusion: The detection of methylated APC and GSTP1 DNA in serum correlated with the presence of CTC in the blood of breast cancer patients. Both methylated DNA and CTC correlated with a more aggressive tumor biology and advanced disease.
机译：背景：外周血中与肿瘤相关的甲基化DNA和循环肿瘤细胞（CTC）可能对乳腺癌的预后具有重要意义。因此，我们研究的目的是检查乳腺癌患者血液中的游离甲基化DNA和CTC，并将其与已知会影响预后的临床病理特征相关联。 -材料和方法：我们前瞻性地从85位乳腺癌患者和22位健康志愿者那里获得了血清样本。通过甲基化特异性PCR（MethyLight PCR）分析血清中的五个基因：腺瘤性息肉病（APC），ras缔合域家族蛋白1A（RASSF1A），雌激素受体1（ESR1），CDKN2A（p16）和谷胱甘肽s-转移酶pi 1 （GSTP1）。 β肌动蛋白（ACTB）作为对照。平行匹配的63名患者的外周血用于通过改良的免疫磁AdnaTest BreastCancerSelect进行外周血循环肿瘤细胞分析，并通过PCR检测EPCAM，MUC1，MGB1和SPDEF。 -结果：APC基因中的高甲基化率为29％（25/85），RASSF1A中为26％（22/85），GSTP1中为18％（14/76），ESR1中为38％（32/85）所有乳腺癌患者中被检测到。没有发现CDKN2A的超甲基化（0/25）。通过检测EPCAM 13％（8/63），MUC1 16％（10/63），MGB 9％（5/55），SPDEF 12％（7/58）和27％来确定患者的血样CTC阳性检测一个或多个基因（15/55）。癌症患者和健康志愿者之间甲基化的APC DNA差异显着。此外，甲基化的APC，RASSF1和CTC在转移性疾病和非转移性疾病中有显着差异。另外，甲基化的APC，RASSF1A和CTC的存在与AJCC分期显着相关（分别为p = 0.001，p = 0.031和0.002）。在健康个体中发现RASSF1和ESR1基因甲基化的发生率很高（均为23％5/22）。甲基化的GSTP1主要存在于原发性大的患者的血清中（p = 0.023），并且与阳性Her2 / neu状态高度相关（p = 0.003）。血清CA15.3升高与甲基化APC和CTC检测高度相关（均p = 0.000）。甲基化的ESR1无法与上述任何参数表现出显着的相关性。外周血中四氯化碳的存在与甲基化的APC（p = 0.012）和甲基化的GSTP1（p = 0.001）显着相关。结论：乳腺癌患者血清中甲基化的APC和GSTP1 DNA的检测与CTC的存在有关。甲基化的DNA和CTC都与更具侵略性的肿瘤生物学和晚期疾病相关。

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