摘要
目的探讨DNA甲基转移酶的表达及抑癌基因RASSF1A的甲基化及两者表达与较大结直肠腺瘤(colorectal adenoma,CA)的关系。方法选取20对直径≥10 mm的CA患者组织及对应瘤旁组织作为对照,分别使用Real-time PCR和Western blot技术检测DNMT1、DNMT3A和DNMT3B mRNA和蛋白表达变化,应用亚硫酸氢盐限制性酶切分析(COBRA)技术分析重复序列LINE-1甲基化水平,应用甲基化特异性PCR(MS-PCR)技术分析抑癌基因RASSF1A的甲基化水平。结果与对照组相比,CA中DNMT3B的mRNA和蛋白水平升高,DNMT3B的活性显著升高。CA组织中LINE-1甲基化水平降低,肿瘤抑制基因RASSF1A的启动子甲基化水平升高,且表达降低。结论DNMT3B过表达可能在CA的发生、发展中起重要作用,具体机制可能与降低整体甲基化水平和升高抑癌基因RASSF1A甲基化水平有关。
Purpose The aim of this study is to investigate the expression of DNA methyltransferase and the methylation of tumor suppressor gene RASSF1A and relationship between their expression and large colorectal adenomas (CA). Methods Twenty pairs of CA samples with a diameter ≥10 mm and corresponding normal colorectal mucosa tissues from patients were used in this study. The expressions of DNMT1, DNMT3A and DNMT3B mRNA and protein were detected by Real-time PCR and Western blot, respectively. Combined bisulfite restriction analysis (COBRA) was used to measure the methylation level of the repeated sequence LINE-1 and methylation specific PCR (MS-PCR) was used to detect the methylation level of the tumor suppressor gene RASSF1A. Results Compared with the control group, the mRNA and protein levels of DNMT3B in CA were increased, and the activity of DNMT3B was significantly in creased. In CA tissues, the level of LINE-1 methylation increased, the promoter methylation level of RASSF1A increased and the RASSF1A expression decreased. Conclusion The overexpression of DNMT3B may play a role in the occurrence and development of CA, and the specific mechanism may be related to reducing the overall level of methylation and increasing the level of RASSF1A.
作者
刘娜
闫少春
杨洁
高永胜
孟宪梅
党彤
樊向文
邵国
LIU Na;YAN Shao-chun;YANG Jie;GAO Yong-sheng;MENG Xian-mei;DANG Tong;FAN Xiang-wen;SHAO Guo(Inner Mongolia Key Laboratory of Hypoxic Translational Medicine,Baotou Medical College,Baotou 014040,China;Department of Pathology,Baotou Central Hospital,Baotou 014040,China;Department of Digestive,Baotou Medical College,Second Affiliated Hospital,Baotou 014030,China;Department of Surgical Oncology,Inner Mongolia Baogang Hospital,Baotou 014010,China)
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2019年第8期887-892,共6页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学基金(81460283、81660307)
内蒙古自然科学研究项目(2018LH08078)
