中华妇产科杂志
中華婦產科雜誌
중화부산과잡지
CHINESE JOUNAL OF OBSTETRICS AND GYNECOLOGY
2012年
3期
196-200
,共5页
仲肇基%杨佳欣%曹冬焱%孙崟%孙璐璐%程雪梅%陈杰%郎景和%沈铿
仲肇基%楊佳訢%曹鼕焱%孫崟%孫璐璐%程雪梅%陳傑%郎景和%瀋鏗
중조기%양가흔%조동염%손음%손로로%정설매%진걸%랑경화%침갱
钙-钙调素依赖性蛋白激酶类%凋亡调节蛋白质类%受体%维甲酸%肿瘤抑制蛋白质类%甲基化%宫颈肿瘤%癌%鳞状细胞
鈣-鈣調素依賴性蛋白激酶類%凋亡調節蛋白質類%受體%維甲痠%腫瘤抑製蛋白質類%甲基化%宮頸腫瘤%癌%鱗狀細胞
개-개조소의뢰성단백격매류%조망조절단백질류%수체%유갑산%종류억제단백질류%갑기화%궁경종류%암%린상세포
Calcium-calmodulin-dependent protein kinases%Apoptosis regulatory proteins%Receptors,retinoic acid%Tumor suppressor proteins%Methylation%Uterine cervical neoplasms%Carcinoma,squamous cell
目的 检测宫颈脱落细胞中DAPK1、RAR-β、MGMT基因启动子的甲基化水平,探讨其筛查高级别宫颈病变[包括宫颈上皮内瘤变(CIN)Ⅱ、Ⅲ]及宫颈鳞癌的价值.方法 采用简单随机法选取2010年3-10月间北京协和医院临床使用后剩余的宫颈脱落细胞标本共103份,其中,细胞学诊断包括正常宫颈细胞12份、不典型鳞状上皮细胞(ASC)17份、低度鳞状上皮内病变(LSIL) 30份、高度鳞状上皮内病变( HSIL)30份、宫颈鳞癌14份；组织学诊断包括正常宫颈组织27份、CIN Ⅰ19份、CIN Ⅱ 17份、CIN Ⅲ 25份、宫颈鳞癌15份.采用甲基化特异性高分辨率熔解曲线分析(MS-HRM)法检测宫颈脱落细胞中DAPK1、RAR-β、MGMT基因启动子的甲基化率,并评价MS-HRM法检测的敏感性；采用第二代杂交捕获技术( HCⅡ)检测宫颈脱落细胞中高危型HPV载量(以HPV-HCⅡ值表示)；评价DAPK1、RAR-β、MGMT基因启动子的甲基化和HCⅡ单独或联合检测筛查高级别宫颈病变及宫颈鳞癌的价值.结果MS-HRM法可以检测出标准样本中的1％DNA甲基化率.在不同细胞学病变中,DAPK1、RAR-β、MGMT基因启动子的甲基化率分别比较,差异均有统计学意义(P值分别为0.000、0.011、0.002)；在不同组织学病变中,DAPK1和RAR-β基因启动子的甲基化率分别比较,差异均有统计学意义(P值分别为0.000、0.021).高级别宫颈病变+宫颈鳞癌与正常宫颈+CINI组织中DAPK1基因启动子的甲基化率分别为1.47％和20.98％,两者比较,差异有统计学意义(P =0.000).DAPK1基因启动子的甲基化检测筛查高级别宫颈病变及宫颈鳞癌的灵敏度为0.571、特异度为0.791,曲线下面积(AUC)为0.709；DAPK1基因启动子的甲基化联合HCⅡ检测的灵敏度为0.825、特异度为0.565,AUC为0.695.结论 MS-HRM法检测宫颈脱落细胞的甲基化水平是可行的.DAPK1、RAR-β基因启动子甲基化可以作为筛查宫颈病变的分子标记.DAPK1基因启动子的甲基化联合HCⅡ检测有助于筛查高级别宫颈病变及宫颈鳞癌.
目的 檢測宮頸脫落細胞中DAPK1、RAR-β、MGMT基因啟動子的甲基化水平,探討其篩查高級彆宮頸病變[包括宮頸上皮內瘤變(CIN)Ⅱ、Ⅲ]及宮頸鱗癌的價值.方法 採用簡單隨機法選取2010年3-10月間北京協和醫院臨床使用後剩餘的宮頸脫落細胞標本共103份,其中,細胞學診斷包括正常宮頸細胞12份、不典型鱗狀上皮細胞(ASC)17份、低度鱗狀上皮內病變(LSIL) 30份、高度鱗狀上皮內病變( HSIL)30份、宮頸鱗癌14份；組織學診斷包括正常宮頸組織27份、CIN Ⅰ19份、CIN Ⅱ 17份、CIN Ⅲ 25份、宮頸鱗癌15份.採用甲基化特異性高分辨率鎔解麯線分析(MS-HRM)法檢測宮頸脫落細胞中DAPK1、RAR-β、MGMT基因啟動子的甲基化率,併評價MS-HRM法檢測的敏感性；採用第二代雜交捕穫技術( HCⅡ)檢測宮頸脫落細胞中高危型HPV載量(以HPV-HCⅡ值錶示)；評價DAPK1、RAR-β、MGMT基因啟動子的甲基化和HCⅡ單獨或聯閤檢測篩查高級彆宮頸病變及宮頸鱗癌的價值.結果MS-HRM法可以檢測齣標準樣本中的1％DNA甲基化率.在不同細胞學病變中,DAPK1、RAR-β、MGMT基因啟動子的甲基化率分彆比較,差異均有統計學意義(P值分彆為0.000、0.011、0.002)；在不同組織學病變中,DAPK1和RAR-β基因啟動子的甲基化率分彆比較,差異均有統計學意義(P值分彆為0.000、0.021).高級彆宮頸病變+宮頸鱗癌與正常宮頸+CINI組織中DAPK1基因啟動子的甲基化率分彆為1.47％和20.98％,兩者比較,差異有統計學意義(P =0.000).DAPK1基因啟動子的甲基化檢測篩查高級彆宮頸病變及宮頸鱗癌的靈敏度為0.571、特異度為0.791,麯線下麵積(AUC)為0.709；DAPK1基因啟動子的甲基化聯閤HCⅡ檢測的靈敏度為0.825、特異度為0.565,AUC為0.695.結論 MS-HRM法檢測宮頸脫落細胞的甲基化水平是可行的.DAPK1、RAR-β基因啟動子甲基化可以作為篩查宮頸病變的分子標記.DAPK1基因啟動子的甲基化聯閤HCⅡ檢測有助于篩查高級彆宮頸病變及宮頸鱗癌.
목적 검측궁경탈락세포중DAPK1、RAR-β、MGMT기인계동자적갑기화수평,탐토기사사고급별궁경병변[포괄궁경상피내류변(CIN)Ⅱ、Ⅲ]급궁경린암적개치.방법 채용간단수궤법선취2010년3-10월간북경협화의원림상사용후잉여적궁경탈락세포표본공103빈,기중,세포학진단포괄정상궁경세포12빈、불전형린상상피세포(ASC)17빈、저도린상상피내병변(LSIL) 30빈、고도린상상피내병변( HSIL)30빈、궁경린암14빈；조직학진단포괄정상궁경조직27빈、CIN Ⅰ19빈、CIN Ⅱ 17빈、CIN Ⅲ 25빈、궁경린암15빈.채용갑기화특이성고분변솔용해곡선분석(MS-HRM)법검측궁경탈락세포중DAPK1、RAR-β、MGMT기인계동자적갑기화솔,병평개MS-HRM법검측적민감성；채용제이대잡교포획기술( HCⅡ)검측궁경탈락세포중고위형HPV재량(이HPV-HCⅡ치표시)；평개DAPK1、RAR-β、MGMT기인계동자적갑기화화HCⅡ단독혹연합검측사사고급별궁경병변급궁경린암적개치.결과MS-HRM법가이검측출표준양본중적1％DNA갑기화솔.재불동세포학병변중,DAPK1、RAR-β、MGMT기인계동자적갑기화솔분별비교,차이균유통계학의의(P치분별위0.000、0.011、0.002)；재불동조직학병변중,DAPK1화RAR-β기인계동자적갑기화솔분별비교,차이균유통계학의의(P치분별위0.000、0.021).고급별궁경병변+궁경린암여정상궁경+CINI조직중DAPK1기인계동자적갑기화솔분별위1.47％화20.98％,량자비교,차이유통계학의의(P =0.000).DAPK1기인계동자적갑기화검측사사고급별궁경병변급궁경린암적령민도위0.571、특이도위0.791,곡선하면적(AUC)위0.709；DAPK1기인계동자적갑기화연합HCⅡ검측적령민도위0.825、특이도위0.565,AUC위0.695.결론 MS-HRM법검측궁경탈락세포적갑기화수평시가행적.DAPK1、RAR-β기인계동자갑기화가이작위사사궁경병변적분자표기.DAPK1기인계동자적갑기화연합HCⅡ검측유조우사사고급별궁경병변급궁경린암.
Objective To assess the correlation of promoter methylation of DAPK1,RAR-β and MGMT with cervical lesions from cytology to histology,and to reveal the clinical value of DNA methylation in diagnosis of cervical intraepithelial neoplasia (CIN).Methods A total of 103 random-selected cervical samples were collected from residual liquid-based cytology specimens after clinical use in cytopathological diagnosis in outpatient clinic of obstetrics and gynecology,Peking Union Medical Collage Hospital from March 2010 to October 2010.Informed consent was obtained from each woman before the initiation of the study.The methylation seusitive-high resolution melt (MS-HRM) assay was used to evaluate promoter methylation of three genes ( DAPKI,RAR-β and MGMT) in 103 biopsy-confirmed liquid-based cervical cytology samples.Methylation levels and high-risk HPV DNA loading ( HC Ⅱ values) were analyzed in relation to both cytological and histological diagnosis.Results The methylation level of all three genes showed significant difference among the different cytological groups ( P =0.000,0.011 and 0.002,respectively).The methylation level of DAPK1 and RAR-β showed significant difference among the different histological groups ( P =0.000 and 0.021 ),while there was no significant difference for MGMT.DAPK1 methylation levels was 1.47％ in the CIN Ⅱ/high-grade precancerous lesions group,and 20.98％ in the normal/CIN I groups ( P =0.000 ),but there was no significant difference between CIN I/high-grade precancerous lesions and normal/CIN Ⅰ groups for RAR-β and MGMT.The combination of DAPK1/HR-HPV loading showed a sensitivity of 0.825 and an area under the receiver operating characteristic curve (ROC) curve (AUC) of 0.695 as diagnostic methods for detecting CIN Ⅱ/high-grade precancerous lesions.Conclusions DNA methylation such as DAPK1 and RAR-β,in combination with HR-HPV detection,may serve as biomarkers to detect CIN Ⅱ/high-grade precancerous lesions.Detection of methylated DNA from liquid-based cervical cytology specimens is technically feasible with the MS-HRM assay.
