子宫颈癌发生发展中维甲酸受体-β基因启动子甲基化状态的研究  

Study on methylation of RAR-β gene promoter in cervical carcinogenesis and development

作　　者：高艳萍 李敏[2] 张颖颖[2] 王翰[2] 贺小红[2] 王泽华[2] 

机构地区：[1]山西省大同市第一人民医院妇科 [2]华中科技大学同济医学院附属协和医院妇产科,武汉430022

出　　处：《肿瘤研究与临床》2008年第4期250-253,共4页Cancer Research and Clinic

摘　　要：目的 研究维甲酸受体-β(RAR-β)基因在子宫颈癌发生、发展中的表达变化,分析其差异表达与甲基化的关系及临床意义.方法 采用反转录聚合酶链反应(RT-PCR)检测四种子宫颈癌细胞株、38例子宫颈浸润癌、30例子宫颈上皮内瘤变组织、10例正常子宫颈组织中RAR-β基因的表达,Western blotting对RAR-β蛋白进行定量检测,分析蛋白水平上的RAR-β表达的变化,并与正常子宫颈组织的RAR-β表达情况进行对照.应用甲基化特异性PCR(MSP)对细胞系和组织的RAR-βDNA进行甲基化分析,同时分析RAR-β基因的表达与子宫颈癌临床病理参数的关系.结果 子宫颈癌细胞系、子宫颈上皮内瘤变组织和子宫颈癌组织中RAR-β基因的表达缺失或下降,并与正常子宫颈组织中的RAR-β表达情况进行对照,差异具有统计学意义(P＜0.05).蛋白的表达检测也得到相同的结果.MSP检测发现3种子宫颈癌细胞株、11例癌组织、1例CIN Ⅰ级、5例CIN Ⅱ～Ⅲ级RAR-β第1外显子甲基化表现,甲基化者RAR-β表达均下降.RAR-β基因甲基化状态在不同的病理分级中差异有统计学意义(P=0.037),在不同的I临床分期(P≈1.000)及有无淋巴转移中差异无统计学意义(P≈1.000).结论 RAR-β基因的表达缺陷在子宫颈癌的发生中起重要作用,而启动子区异常甲基化是导致基因表达缺陷的主要原因.Objective To study the expressive changes of RAR-β gene in cervical carcinogenesis and analyze the relation between this differentiated expression and methylation, and discuss the clinical significance of this differentiated expression.Methods RT-PCR method was used to detect the expression of RAR-β in 4 cervical eancer cell lines,38 eases of cevical invasive carcinomatous tissues,30 cases of cevical intraepithelial neoplasia,and 10 cases of normal cervical tissues.Using Western Blot method to detect the expression of RAR-β protein quantificationally, analyze the RAR-β gene expressive difference in protein level.Using the methylation-*specific PCR methods to analyze the RAR-β methylation state in cell lines and tissues.Meanwhile analyze the relationship between the RAR-β gene expression and the clinical pathological parameters.Resuits The expression of RAR-β gene was deleted or deereaseed in cervieal cancer celllines,cevical intraepithelial neoplasia and cevical carcinoma,which was significantly difierent from normal tissues(P＜0.05).The same results was found in the protein expression.The methylation in RAR-β gene first exon was detected in 3 types of cell lines,11 cases of carcinoma,1 case of CIN Ⅰ and 5 cases of CIN Ⅱ～Ⅲ.while the RAR-β gene expression was decreased in those cell lines and tissues.There was significant differenee of RAR-β gene methylation in different pathological grades (P=0.037).There was no statistical significant difference of clinical stages(P≈1.000)and the metastasis of lymph node(P≈1.000).Conclusion The defective expression of RAR-β gene plays an important role in the carcinogenesis of cervical cancer.The abnormal methylation of promoter region was the main cause of defective expression of RAR-β gene.

关 键 词：子宫颈肿瘤 受体 维甲酸 甲基化 

分 类 号：R737[医药卫生—肿瘤]