中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2013年
8期
600-603
,共4页
忙尼沙汗·阿不都拉%古丽那尔·阿布都拉江%热沙来提·艾米多%阿布力孜·阿布杜拉%阿仙姑·哈斯木
忙尼沙汗·阿不都拉%古麗那爾·阿佈都拉江%熱沙來提·艾米多%阿佈力孜·阿佈杜拉%阿仙姑·哈斯木
망니사한·아불도랍%고려나이·아포도랍강%열사래제·애미다%아포력자·아포두랍%아선고·합사목
宫颈肿瘤%宫颈上皮内瘤样病变%DNA甲基化%内质网蛋白57%维吾尔族
宮頸腫瘤%宮頸上皮內瘤樣病變%DNA甲基化%內質網蛋白57%維吾爾族
궁경종류%궁경상피내류양병변%DNA갑기화%내질망단백57%유오이족
Uterine cervical neoplasms%Cervical intraepithelial neoplasms%DNA methylation%Endoplasmic reticulum protein 57%Uyghur
目的 探讨宫颈上皮内瘤变和宫颈癌与内质网蛋白57(ERp57)基因启动子区甲基化水平的关系和意义.方法 应用MethPrimers软件设计ERp57基因启动子区CpG岛特异性PCR引物,对宫颈癌SiHa细胞DNA进行亚硫酸氢盐修饰、目的片段扩增、质粒载体克隆和测序,确定目的片段所含CpG序列甲基化情况.收集15例正常宫颈上皮(对照组)、30例宫颈上皮内瘤变(CIN组)和33例宫颈鳞癌(CSCC组)患者的新鲜组织标本,应用基质辅助激光解吸电离飞行时间质谱(MALDITOF)技术定量分析宫颈病变组织DNA ERp57基因启动子单一CpG位点的甲基化水平.结果 ERp57基因相应目的片段各含9个CpG位点,在宫颈癌SiHa细胞基因组DNA中,所有的CpG位点都发生了甲基化,甲基化率为100％.MALDI-TOF检测结果显示,在CSCC组中,CpG_1、CpG_5和CpG_7位点的甲基化水平分别为0.9021±0.1810、0.8521±0.1696和0.9671 ±0.1425,对照组分别为0.7721±0.0644、0.7347±0.0924和0.8765±0.0226,差异均有统计学意义(均P＜0.05).CSCC组、CIN组和对照组宫颈组织ERp57基因的甲基化水平分别为0.9065±0.3709、0.8697±0.3336和0.8040±0.4705,差异无统计学意义(P =0.052).结论 维吾尔族妇女宫颈病变组织中,ERp57基因启动子某些单个CpG位点发生甲基化后,引起该基因转录下调,其位点可能是关键性CpG位点.
目的 探討宮頸上皮內瘤變和宮頸癌與內質網蛋白57(ERp57)基因啟動子區甲基化水平的關繫和意義.方法 應用MethPrimers軟件設計ERp57基因啟動子區CpG島特異性PCR引物,對宮頸癌SiHa細胞DNA進行亞硫痠氫鹽脩飾、目的片段擴增、質粒載體剋隆和測序,確定目的片段所含CpG序列甲基化情況.收集15例正常宮頸上皮(對照組)、30例宮頸上皮內瘤變(CIN組)和33例宮頸鱗癌(CSCC組)患者的新鮮組織標本,應用基質輔助激光解吸電離飛行時間質譜(MALDITOF)技術定量分析宮頸病變組織DNA ERp57基因啟動子單一CpG位點的甲基化水平.結果 ERp57基因相應目的片段各含9箇CpG位點,在宮頸癌SiHa細胞基因組DNA中,所有的CpG位點都髮生瞭甲基化,甲基化率為100％.MALDI-TOF檢測結果顯示,在CSCC組中,CpG_1、CpG_5和CpG_7位點的甲基化水平分彆為0.9021±0.1810、0.8521±0.1696和0.9671 ±0.1425,對照組分彆為0.7721±0.0644、0.7347±0.0924和0.8765±0.0226,差異均有統計學意義(均P＜0.05).CSCC組、CIN組和對照組宮頸組織ERp57基因的甲基化水平分彆為0.9065±0.3709、0.8697±0.3336和0.8040±0.4705,差異無統計學意義(P =0.052).結論 維吾爾族婦女宮頸病變組織中,ERp57基因啟動子某些單箇CpG位點髮生甲基化後,引起該基因轉錄下調,其位點可能是關鍵性CpG位點.
목적 탐토궁경상피내류변화궁경암여내질망단백57(ERp57)기인계동자구갑기화수평적관계화의의.방법 응용MethPrimers연건설계ERp57기인계동자구CpG도특이성PCR인물,대궁경암SiHa세포DNA진행아류산경염수식、목적편단확증、질립재체극륭화측서,학정목적편단소함CpG서렬갑기화정황.수집15례정상궁경상피(대조조)、30례궁경상피내류변(CIN조)화33례궁경린암(CSCC조)환자적신선조직표본,응용기질보조격광해흡전리비행시간질보(MALDITOF)기술정량분석궁경병변조직DNA ERp57기인계동자단일CpG위점적갑기화수평.결과 ERp57기인상응목적편단각함9개CpG위점,재궁경암SiHa세포기인조DNA중,소유적CpG위점도발생료갑기화,갑기화솔위100％.MALDI-TOF검측결과현시,재CSCC조중,CpG_1、CpG_5화CpG_7위점적갑기화수평분별위0.9021±0.1810、0.8521±0.1696화0.9671 ±0.1425,대조조분별위0.7721±0.0644、0.7347±0.0924화0.8765±0.0226,차이균유통계학의의(균P＜0.05).CSCC조、CIN조화대조조궁경조직ERp57기인적갑기화수평분별위0.9065±0.3709、0.8697±0.3336화0.8040±0.4705,차이무통계학의의(P =0.052).결론 유오이족부녀궁경병변조직중,ERp57기인계동자모사단개CpG위점발생갑기화후,인기해기인전록하조,기위점가능시관건성CpG위점.
Objective To investigate the relationship and significance between endoplasmic reticulum protein 57 (ERp57) gene promoter region methylation with the pathogenesis of cervical lesions in Uighur women.Methods The special software was used to design specific primers of CpG island fragments of ERp57 gene promoter and bisulfite-modified SiHa cancer cell DNA for PCR amplification,cloning and sequencing the target fragments to obtain relevant information of CpG methylation in the gene base sequencs.Seventy-eight fresh tissues of CIN,CSCC and normal control were collected,and the methylation level of ERp57 gene promoter regions in different cervical lesions were identified using Sequenom MassARRAY(R)DNA technology.Results ERp57 gene corresponding target fragment contained the 18 CpG sites.All of the CpG sites methylation occurred in SiHa cervical cancer cell genomic DNA.The analysis of the data resulted from the quantitative analysis of single CpG site methylation by Sequenom MassARRAY platform showed that the methylation level between three CpG sites (CpG_1,CpG_5 and CpG_7) from CpG_1,CpG_2,CpG_3.4,CpG_5,CpG_6,CpG_7,CpG_8 and CpG_9 had significant differences in the CSCC,CIN or control groups.Conclusions Although the global methylation level of the ERp57 gene promoter is higher in CSCC than that in CIN and normal control tissues in Uighur women,hypermethylation occurs only in certain CpG islands and sites.This indicates that the regulation of expression by DNA methylation is not CpG islandspecific,but varies for individual CpG sites,and may explain to a certain extent the epigenetic mechanisms regulated by Erp57 gene expression.
