中华病理学杂志
中華病理學雜誌
중화병이학잡지
Chinese Journal of Pathology
2012年
7期
438-442
,共5页
古丽扎热叶·艾库拉%焦桢%吴小川%马俊旗%阿布力孜·阿布杜拉%阿仙姑·哈斯木
古麗扎熱葉·艾庫拉%焦楨%吳小川%馬俊旂%阿佈力孜·阿佈杜拉%阿仙姑·哈斯木
고려찰열협·애고랍%초정%오소천%마준기%아포력자·아포두랍%아선고·합사목
维吾尔族%宫颈疾病%DNA甲基化%乳头状瘤病毒,人
維吾爾族%宮頸疾病%DNA甲基化%乳頭狀瘤病毒,人
유오이족%궁경질병%DNA갑기화%유두상류병독,인
Uygur nationality%Cervix diseases%DNA methylation%Papillomavirus,human
目的 探讨维吾尔族妇女宫颈病变及人乳头状瘤病毒(HPV)感染与抗原处理相关转运体(TAP)基因启动子区甲基化水平的关系和意义.方法 利用专业软件设计TAP1和TAP2基因启动子区含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,确定该区域所含CpG序列的甲基化情况.收集维吾尔族妇女15例正常宫颈上皮、30例宫颈鳞状上皮内肿瘤形成(CIN)和33例宫颈鳞癌患者的新鲜组织标本共78例,提取DNA,采用MassArray DNA技术平台(质谱分析)定量分析TAP1和TAP2基因启动子甲基化水平,同时以HPV分型芯片鉴定HPV亚型,分析基因甲基化与HPV感染的关系.结果 (1)TAP1和TAP2基因相应目的片段各含23个和8个CpG位点,在宫颈癌SiHa细胞基因组DNA中,分别有5个和8个CpG位点发生甲基化.宫颈病变病理过程伴随着TAP1基因CpG片段甲基化水平的改变,其在宫颈鳞癌和CIN组织的甲基化率[(0.048±0.039)和(0.037±0.026)]高于正常宫颈上皮组织(0.035±0.029),差异有统计学意义(P<0.05).尽管TAP2基因甲基化水平也发生了相应改变,但其差异无统计学意义(P>0.05).HPV分型芯片检出13种HPV基因型,其中HPV16所占比率为66.7%(52/78),HPV16亚型阳性与宫颈病变进程及TAP1基因甲基化率的升高趋势呈正相关(x2=6.08,P=0.039).结论 TAP1基因启动子区CpG岛甲基化是宫颈癌病变的一种特异性改变,与HPV16感染可能存在密切关系.
目的 探討維吾爾族婦女宮頸病變及人乳頭狀瘤病毒(HPV)感染與抗原處理相關轉運體(TAP)基因啟動子區甲基化水平的關繫和意義.方法 利用專業軟件設計TAP1和TAP2基因啟動子區含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,確定該區域所含CpG序列的甲基化情況.收集維吾爾族婦女15例正常宮頸上皮、30例宮頸鱗狀上皮內腫瘤形成(CIN)和33例宮頸鱗癌患者的新鮮組織標本共78例,提取DNA,採用MassArray DNA技術平檯(質譜分析)定量分析TAP1和TAP2基因啟動子甲基化水平,同時以HPV分型芯片鑒定HPV亞型,分析基因甲基化與HPV感染的關繫.結果 (1)TAP1和TAP2基因相應目的片段各含23箇和8箇CpG位點,在宮頸癌SiHa細胞基因組DNA中,分彆有5箇和8箇CpG位點髮生甲基化.宮頸病變病理過程伴隨著TAP1基因CpG片段甲基化水平的改變,其在宮頸鱗癌和CIN組織的甲基化率[(0.048±0.039)和(0.037±0.026)]高于正常宮頸上皮組織(0.035±0.029),差異有統計學意義(P<0.05).儘管TAP2基因甲基化水平也髮生瞭相應改變,但其差異無統計學意義(P>0.05).HPV分型芯片檢齣13種HPV基因型,其中HPV16所佔比率為66.7%(52/78),HPV16亞型暘性與宮頸病變進程及TAP1基因甲基化率的升高趨勢呈正相關(x2=6.08,P=0.039).結論 TAP1基因啟動子區CpG島甲基化是宮頸癌病變的一種特異性改變,與HPV16感染可能存在密切關繫.
목적 탐토유오이족부녀궁경병변급인유두상류병독(HPV)감염여항원처리상관전운체(TAP)기인계동자구갑기화수평적관계화의의.방법 이용전업연건설계TAP1화TAP2기인계동자구함CpG도특이성PCR인물,대궁경암SiHa세포DNA진행아류산경염수식、목적편단확증、질립재체극륭화측서,학정해구역소함CpG서렬적갑기화정황.수집유오이족부녀15례정상궁경상피、30례궁경린상상피내종류형성(CIN)화33례궁경린암환자적신선조직표본공78례,제취DNA,채용MassArray DNA기술평태(질보분석)정량분석TAP1화TAP2기인계동자갑기화수평,동시이HPV분형심편감정HPV아형,분석기인갑기화여HPV감염적관계.결과 (1)TAP1화TAP2기인상응목적편단각함23개화8개CpG위점,재궁경암SiHa세포기인조DNA중,분별유5개화8개CpG위점발생갑기화.궁경병변병리과정반수착TAP1기인CpG편단갑기화수평적개변,기재궁경린암화CIN조직적갑기화솔[(0.048±0.039)화(0.037±0.026)]고우정상궁경상피조직(0.035±0.029),차이유통계학의의(P<0.05).진관TAP2기인갑기화수평야발생료상응개변,단기차이무통계학의의(P>0.05).HPV분형심편검출13충HPV기인형,기중HPV16소점비솔위66.7%(52/78),HPV16아형양성여궁경병변진정급TAP1기인갑기화솔적승고추세정정상관(x2=6.08,P=0.039).결론 TAP1기인계동자구CpG도갑기화시궁경암병변적일충특이성개변,여HPV16감염가능존재밀절관계.
Objective To study the relationship between TAP(transporter associated with antigen processing) gene promoter regional methylation level and cervical lesions with HPV infection in Uyghur women.Methods A specialized software was used to design specific primers of CpG island fragments of TAP1 and TAP2 gene promoter for PCR amplification,bisulfitemodified SiHa cancer cell DNA for PCR amplification,cloning and sequencing analysis to obtain the relevant information on the gene base sequence methylation of CpG sites.Seventy-eight fresh cervical tissue samples from Uyghur women with cervicitis (number = 15),cervical intraepithelial neoplasia(CIN,number = 30) and cervical squamous cell carcinoma(number = 33) were collected.The methylation level of TAP1 and TAP2 gene promoter regions was detected using MassArray DNA technology.HPV infection status was determined by HPV gene chips.The relationship between CpG-island methylation of gene promoter regions and HPV infection was then analyzed.Results Each TAP1 and TAP2 gene corresponding target fragment contained 23 and 8 CpG sites.There were 5 and 8 CpG sites methylation occurred in SiHa cervical cancer cells genomic DNA respectively.The TAP1 methylation level increased steadily with the severity of cervical lesions.The methylation levels in cervical aquamous cell carcinoma and CIN(0.048 4±0.039 and 0.037 4±0.026,respectively) were higher than that of normal cervical tissue(0.035 ± 0.029,P <0.05).Although TAP2 gene methylation level also demonstrated similar changes,the difference however was not statistically significant(P > 0.05).HPV gene chip deteeted 13 HPV genotypes,with HPV16 infection rate being 66.7%(52/78).The methylated proportion of TAP1 positively correlated with HPV16 infection(x2 =6.08,P =0.039).Conclusion TAP1 methylation is a remarkable phenomenon occurring in a range of cervical lesions and significantly associated with cervical HPV infection.