Detection of multiple gene hypermethylation in the development of esophageal squamous cell carcinoma.
Detection of multiple gene hypermethylation in the development of esophageal squamous cell carcinoma.
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DOI：
10.1093/carcin/23.10.1713
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发表时间：
2002-10
期刊：
影响因子：
4.7
通讯作者：
Y. Nie;J. Liao;Xin Zhao;Yunlong Song;Guang-Yu Yang;Li‐dong Wang;Chung S. Yang
中科院分区：
文献类型：
--
作者：
Y. Nie;J. Liao;Xin Zhao;Yunlong Song;Guang-Yu Yang;Li‐dong Wang;Chung S. Yang
与肿瘤抑制基因相关的CpG岛异常高甲基化可导致基因表达抑制，并显着促进肿瘤发生。食管鳞状细胞癌（ESCC）是一种多阶段的恶性肿瘤，包括基底细胞增生（BCH）、异型增生（DYS）、原位癌（CIS）和癌变。在本研究中，我们研究了10个选定的基因在正常人和食管鳞癌患者切除组织的活检组织中的超甲基化。采用激光捕获显微切割技术，从切除的组织中分离肿瘤和邻近的正常组织和癌前组织，包括BCH、DYS和CIS。在这些样本中检测了10个基因的CpG岛的超甲基化：p16（INK 4 a）、p15（INK 4 B）、p14（ARF）、人类白细胞抗原（HLA）-A、-B、-C、hMLH 1、E-钙粘蛋白（E-cad）、脆性组氨酸三联体和von Hippel-Lindau（VHL）。分别与E-cad和VHL相邻的两个Alu序列的甲基化被用作对照，以验证DNA提取和化学修饰的过程。在48例BCH或DYS活检标本中，最常见的高甲基化基因是p16（INK 4a）（18.8%）和p14（ARF）（14.6%）。这48个样本中有17个（35.4%）含有至少一个基因的超甲基化。在切除的组织中，52%的BCH和81%的肿瘤显示至少一个基因的高甲基化。在同一患者的早期病变中高甲基化的基因总是在晚期病变中被发现高甲基化。所有基因在某些阶段都发生了甲基化，并根据其频率将其聚为四类。第一组基因由p16（INK 4a）和p14（ARF）组成，在所有阶段中最频繁地高甲基化，并且频率从正常上皮（0%）到BCH，再到乳腺癌/原位癌和ESCC增加。其他基因的高甲基化频率较低。我们的研究结果表明，关键基因，如p16（INK 4a），p14（ARF）和hMLH 1的超甲基化，可与其他分子变化，如p53突变，在发展生物标志物预测ESCC的风险。
Abnormal hypermethylation of CpG islands associated with tumor suppressor genes can lead to repression of gene expression and contribute significantly to tumorigenesis. Esophageal squamous cell carcinoma (ESCC) is thought to be developed through a multi-stage process, which involves basal cell hyperplasia (BCH), dysplasia (DYS), carcinoma in situ (CIS) and carcinoma. In the present study, we studied the hypermethylation of 10 selected genes in biopsies from normal individuals and resected tissues from ESCC patients. Tumor and neighboring normal and precancerous tissues including BCH, DYS and CIS were microdissected from the resected tissues by laser capture microdissection. Hypermethylation of CpG islands was examined in these samples for 10 genes: p16(INK4a), p15(INK4b), p14(ARF), human leukocyte antigen (HLA)-A, -B, -C, hMLH1, E-cadherin (E-cad), fragile histidine triad and von Hippel-Lindau (VHL). Methylation of two Alu sequences, which neighbor E-cad and VHL, respectively, was used as control to verify the procedure of DNA extraction and chemical modification. In 48 biopsy samples with BCH or DYS, the most frequent hypermethylated genes were p16(INK4a) (18.8%) and p14(ARF) (14.6%). Seventeen out of these 48 samples (35.4%) contained hypermethylation of at least one gene. In the resected tissues, 52% of the BCH and 81% of the tumors showed hypermethylation of at least one gene. Genes hypermethylated in earlier stage lesions were always found hypermethylated at the later stage lesions in the same patient. All of the genes were methylated at some stages and they were clustered into four groups according to their frequencies. The first group of genes, which consisted of p16(INK4a) and p14(ARF), was most frequently hypermethylated in all stages, and the frequencies increased from normal epithelial (0%) to BCH, to displasia/carcinoma in situ and ESCC. Other genes were hypermethylated less frequently. Our results suggest that hypermethylation of key genes, such as p16(INK4a), p14(ARF) and hMLH1, may be used in combination with other molecular changes, such as p53 mutation, in the development of biomarkers for predicting the risk for ESCC.