宫颈癌大鼠模型癌组织中DAPK基因启动子甲基化与宫颈癌发展和转移的关联  

Association of DAPK gene promoter methylation in cervical cancer rat model with the development and metastasis of cervical cancer

作　　者：孙晓东[1] 任娟娟 康婷[1] 胡海峰[1] SUN Xiao-dong;REN Juan-juan;KANG Ting(Second Ward,Department of Oncology,Affiliated Hospital of Yan'an University,Yan'an Shaanxi 716000,China;Department of Obstetrics and Gynecology,Second Hospital of Yulin City,Yulin Shaanxi 719000,China)

机构地区：[1]延安大学附属医院肿瘤科二病区,陕西延安716000 [2]榆林市第二医院妇产科,陕西榆林719000

出　　处：《临床和实验医学杂志》2021年第20期2140-2144,共5页Journal of Clinical and Experimental Medicine

基　　金：陕西省卫生厅课题(编号:2021A014)。

摘　　要：目的探讨宫颈癌大鼠模型癌组织中DAPK基因启动子甲基化与宫颈癌发展和转移的关联。方法选择45只成年雌性Sprague-Dawley大鼠为研究对象,通过皮下注射宫颈癌SiHa细胞建立异种移植瘤动物模型。选取其中35只大鼠进行甲基化实验分组:对照组(正常大鼠,腹腔注射DMSO,n=10),宫颈癌组(宫颈癌大鼠模型,n=25)和宫颈癌邻近组(与宫颈癌组为相同大鼠,获取癌旁组织样本,n=25),通过甲基化特异性聚合酶链式反应分析各组大鼠组织的DAPK甲基化状态;实时荧光定量聚合酶链式反应(RT-PCR)分析大鼠组织中DAPK mRNA的表达。同时将剩余10只宫颈癌大鼠根据实验目的分为宫颈癌组(宫颈癌大鼠模型,给予DMSO腹腔注射,n=5)和甲基化抑制组(在宫颈癌大鼠的模型中注射1 mg/g 5-Aza-CdR,n=5),每周1次,注射至第40天,游标卡尺测量并计算异种移植大鼠肿瘤体积生长,并对肿瘤组织进行称重分析;RT-PCR分析大鼠组织中DAPK mRNA的表达。结果对照组大鼠未出现甲基化,宫颈癌组有19只大鼠肿瘤出现了DAPK甲基化,甲基化率为76.00%,宫颈癌邻近组4只出现DAPK甲基化,甲基化率为16.00%,宫颈癌组较对照组和宫颈癌邻近组的DAPK甲基化率升高,宫颈癌邻近组较对照组DAPK甲基化率升高,差异有统计学意义(P<0.05)。在任何具有启动子甲基化的肿瘤组织或邻近组织中均未检测到DAPK mRNA的表达,在所有启动子未甲基化的肿瘤组织、肿瘤邻近组织和正常组织中均检测到DAPK mRNA的表达,甲基化组织DAPK mRNA的表达显著低于未甲基化组织,差异有统计学意义(P<0.05)。注射后第10天肿瘤体积、重量比较差异无统计学意义(P>0.05),注射后第20、30、40天甲基化抑制组较宫颈癌组大鼠肿瘤体积和重量减小,差异有统计学意义(P<0.05)。仅用PBS处理的大鼠的肿瘤组织中都没有发现DAPK mRNA表达,而在来自用5-Aza-CdR处理的大鼠的所有肿瘤中检测到DAPK mRNA的重新Objective To explore the relationship between DAPK gene promoter methylation in cervical cancer rat model cancer tissue and the development and metastasis of cervical cancer.Methods Forty-five adult female Sprague-Dawley rats were selected as the research objects,and xenograft animal models were established by subcutaneous injection of cervical cancer SiHa cells.35 rats were selected for methylation experiment grouping:control group(normal rats,intraperitoneal injection of DMSO,n=10),cervical cancer group(cervical cancer rat model,n=25)and the cervical cancer adjacent group(the same rats as the cervical cancer group,get samples of adjacent tissues,n=25).The methylation status of DAPK in rat tissues of each group was analyzed by methylation-specific polymerase chain reaction,and the expression of DAPK mRNA in rat tissues was analyzed by real-time polymerase chain reaction(RT-PCR).At the same time,the remaining 10 cervical cancer rats were divided into cervical cancer group(cervical cancer rat model,intraperitoneal injection of DMSO,n=5)and methylation suppression group(inject 1 mg/g 5-Aza-CdR in a rat model of cervical cancer,n=5)according to the experimental purpose,once a week,to the 40th day,the vernier caliper was used to measure and calculate the tumor volume growth of xenotransplanted rats,and the tumor tissue was weighed and analyzed,and the expression of DAPK mRNA in rat tissues was analyzed by RT-PCR.Results There was no methylation in the control group,19 rats in the cervical cancer group showed DAPK methylation,the methylation rate was 76.00%,and 4 rats in the adjacent cervical cancer group showed DAPK methylation and methylation.The rate of DAPK methylation in the cervical cancer group was 16.00%higher than that of the control group and the cervical cancer adjacent group,and the DAPK methylation rate of the cervical cancer adjacent group was higher than that of the control group,the difference was statistically significant(P<0.05).DAPK mRNA expression was not detected in any tumor tissues or adjacent t

关 键 词：宫颈癌 DAPK 甲基化 异种移植 肿瘤生长 

分 类 号：R737.33[医药卫生—肿瘤]