Yan Wen-Ji, Guo Ming-Zhou, Yang Yun-Sheng
Department of Gastroenterology & Hepatology, Chinese PLA General Hospital, Beijing 100853, China.
Zhonghua Nei Ke Za Zhi. 2012 May;51(5):390-3.
To study the association of ubiquitin carboxyl-terminal hydrolase L1 (UCHL1) promoter region methylation with human esophageal cancer.
Promoter region methylation of UCHL1 was detected by methylation specific PCR (MSP) in esophageal cancer cell lines and tissue samples. The expression of UCHL1 was detected by semi-quantitative RT-PCR and Western blot in esophageal cancer cell lines. 5-Aza-2'-deoxycytidine (5-Aza) was applied to reactivate methylated cell lines.
Complete methylation of UCHL1 promoter region was detected in 8 cell lines (KYSE30, KYSE150, KYSE140, KYSE450, KYSE510, TE3, TE7, TE10). Loss of UCHL1 expression was found in 7 cell lines (KYSE30, KYSE150, KYSE140, KYSE450, KYSE510, TE3, TE7). Reduced expression was found in TE10 cell line. Promoter region hypermethylation was correlated with UCHL1 expression in esophageal cancer cell lines. Re-expression of UCHL1 was induced by 5-Aza treatment in KYSE150 and TE3 cell lines. UCHL1 was frequently methylated in human primary esophageal cancer (74.51%, 38/51), while no methylation was detected in normal esophageal mucosa(0/10). No association was found between promoter region methylation and age, gender, tumor location, tumor stage or lymph node metastasis.
UCHL1 is silenced by promoter region hypermethylation in human esophageal cancer. Methylation of UCHL1 is frequently happened to primary esophageal cancer and may play an important role in the tumorigenesis.
研究泛素羧基末端水解酶L1（UCHL1）启动子区域甲基化与人类食管癌的关系。
采用甲基化特异性PCR（MSP）检测食管癌细胞系和组织样本中UCHL1启动子区域的甲基化情况。通过半定量逆转录PCR和蛋白质免疫印迹法检测食管癌细胞系中UCHL1的表达。应用5-氮杂-2'-脱氧胞苷（5-Aza）使甲基化的细胞系重新激活。
在8个细胞系（KYSE30、KYSE150、KYSE140、KYSE450、KYSE510、TE3、TE7、TE10）中检测到UCHL1启动子区域完全甲基化。在7个细胞系（KYSE30、KYSE150、KYSE140、KYSE450、KYSE510、TE3、TE7）中发现UCHL1表达缺失。在TE10细胞系中发现表达降低。食管癌细胞系中启动子区域高甲基化与UCHL1表达相关。5-Aza处理可诱导KYSE150和TE3细胞系中UCHL1重新表达。UCHL1在人类原发性食管癌中频繁甲基化（74.51%，38/51），而在正常食管黏膜中未检测到甲基化（0/10）。未发现启动子区域甲基化与年龄、性别、肿瘤位置、肿瘤分期或淋巴结转移之间存在关联。
在人类食管癌中，UCHL1因启动子区域高甲基化而沉默。UCHL1甲基化在原发性食管癌中频繁发生，可能在肿瘤发生中起重要作用。
