宫颈腺癌与鳞癌组织基因甲基化测定  

Determination of gene methylation in cervical adenocarcinoma and squamous cell carcinoma

作　　者：袁敏[1] 姚立丽[1] 袁建林[1] 古扎丽努尔·阿不力孜[1] YUAN Min;YAO Li-li;YUAN Jian-lin;GUZHALINUER·Abulizi(Department of Gynecology,Affiliated Tumour Hospital of Xinjiang Medical University,Ulumuqi 830011,P.R.China)

机构地区：[1]新疆医科大学附属肿瘤医院妇科,新疆乌鲁木齐830011

出　　处：《中华肿瘤防治杂志》2020年第20期1643-1650,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：新疆维吾尔自治区自然科学基金面上项目(2017D01C396)。

摘　　要：目的宫颈癌变过程中包括表观基因与调控基因的改变,以DNA甲基化为代表的表观遗传改变可导致宫颈癌的发生。本研究通过分析基因甲基化在早期宫颈腺癌组织中的表达差异性,寻找预测宫颈腺癌发生发展的分子标志物。方法收集新疆医科大学附属肿瘤医院2015-01-01-2017-06-30保存的新鲜初治原发宫颈腺癌组织15例和宫颈鳞状细胞癌组织15例,以及同期住院的正常宫颈组织15例,应用Illumina 850K甲基化芯片筛选宫颈腺癌组织的特异甲基化表达基因,亚硫酸盐测序法(bisulfite sequencing PCR,BSP)及实时定量RT-PCR(quantitative real time RT-PCR,Q-RT-PCR)方法对宫颈腺癌组织、宫颈鳞状细胞癌组织及正常宫颈组织进行验证。结果芯片筛选结果显示,性别决定区Y框蛋白1(sex determining region Y-box1,SOX1)、细胞周期蛋白D1(Cyclin D1,CCND1)和蛋白激酶Cα(protein kinase Cα,PRKCA)等基因是宫颈腺癌组织中基因调控网络的中心节点。BSP检测结果显示,SOX1基因甲基化程度在宫颈腺癌为(0.975±0.013)%,宫颈鳞癌为(0.871±0.073)%,正常宫颈组织为(0.416±0.256)%,差异有统计学意义,F=18.640,P<0.001。SOX1是宫颈腺癌特异甲基化表达基因。CCND1(F=0.890,P=0.436)和PRKCA(F=1.749,P=0.693)不是宫颈腺癌特异甲基化表达基因,差异均无统计学意义。Q-RT-PCR检测结果发现,SOX1(t=3.213,P=0.009)、PRKCA(t=-3.448,P=0.006)在宫颈腺癌组与宫颈鳞癌组比较差异均有统计学意义。SOX1(t=2.190,P=0.043)、PRKCA(t=-2.561,P=0.028)在宫颈腺癌组与正常宫颈组比较差异均有统计学意义,CCND1在宫颈腺癌组与正常宫颈组(t=3.937,P=0.003)、宫颈鳞癌组与正常宫颈组(t=4.100,P=0.002)差异均有统计学意义。结论抑癌基因SOX1是宫颈腺癌特异甲基化表达基因,有望成为宫颈腺癌诊断的特异标志物。CCND1、PRKCA基因不能证明是宫颈腺癌特异甲基化表达基因,考虑与BSP检测样本量少、检测位点少有关,有待�OBJECTIVE This study aimed to analyze the difference of gene methylation in early cervical adenocarcinoma and to find molecular markers for predicting the occurrence and development of cervical adenocarcinoma.METHODS We collected 15 fresh primary cervical adenocarcinoma tissues,15 squamous cell carcinoma tissues and 15 normal cervical tissues of Tumor Hospital Affiliated to Xinjiang Medical University from January 1,2015 to June 30,2017.A total of 15 cases of primary cervical adenocarcinoma and 15 cases of primary cervical squamous cell carcinoma at stagesⅠB1 orⅡA1 were included in the study.Infinium Methylation EPIC BeadChip(850 K)was used to screen specifically expressed genes in cervical adenocarcinoma tissues.Bisulfite sequencing polymerase chain reaction(BSP)and quantitative real-time polymerase chain reaction were used to verify the methylation levels in cervical adenocarcinoma,cervical squamous cell carcinoma and normal cervical tissues.RESULTS SOX1,CCND1 and PRKCAgenes participated in multiple signaling pathways,being the central nodes of gene regulatory networks.BSP results showed that the methylation level of SOX1 gene was(0.975±0.013)%in cervical adenocarcinoma,(0.871±0.073)%in cervical squamous cell carcinoma and(0.416±0.256)%in normal cervical tissue.The difference was statistically significant among the three groups(F=18.640,P<0.001),SOX1 was the methylation gene of cervical adenocarcinoma.CCND1(F=0.890,P=0.436)and PRKCA(F=1.749,P=0.693)were not specific methylation genes of cervical adenocarcinoma,and there was no significant difference between them.The results of Q-RT-PCR showed that SOX1(t=3.213,P=0.009)and PRKCA(t=-3.448,P=0.006)were significantly different between cervical adenocarcinoma group and cervical squamous cell carcinoma group.The differences of SOX1(t=2.190,P=0.043),PRKCA(t=-2.561,P=0.028)between cervical adenocarcinoma group and normal cervical group were statistically significant.The differences of CCND1 between cervical adenocarcinoma group and normal cervical group(t=3.937,

关 键 词：宫颈腺癌 SOX1 CCND1 PRKCA 甲基化 

分 类 号：R737.33[医药卫生—肿瘤]