遗传性卵巢癌组织中BRCA1基因杂合性丢失的研究

目的：探讨乳腺癌基因（ＢＲＣＡ１）在遗传性卵巢癌发生、发展中的作用,并分析ＢＲＣＡ１ 基因杂合性丢失（ＬＯＨ）。方法：应用多聚合酶链反应技术（ＰＣＲＴ）,对１０份遗传性卵巢癌组织中ＢＲＣＡ１ 基因内部的Ｄ１７Ｓ８５５ 微卫星位点进行ＬＯＨ检测。结果：２份发生杂合性丢失,ＬＯＨ发现率为２０％ 。结论：ＢＲＣＡ１在遗传性卵巢癌中具有肿瘤抑制基因的作用,同时不能排除其他肿瘤抑制基因的作用。     

卵巢上皮性肿瘤的临床病理特征及其细胞周期素D1和p53蛋白表达的研究

目的研究卵巢上皮性癌（卵巢癌）和交界性上皮性肿瘤的临床病理特征及其细胞周期素D1（cyclin D1）和p53蛋白表达的情况,探讨卵巢癌和交界性上皮性肿瘤在发病机制上的联系。方法分析45例卵巢癌（卵巢癌组）和54例卵巢交界性上皮性肿瘤（交界性肿瘤组）的临床病理资料,采用免疫组化法检测两组组织中cyclin D1、p53蛋白的表达情况,并分析其与临床病理特征的相关性。结果（1）临床病理特征：①年龄：交界性肿瘤组平均年龄为42．5岁（14～82岁）,中位数年龄41岁;卵巢癌组平均年龄为53．5岁（26～80岁）,中位数年龄51岁。②分期：按国际妇产科联盟（FIGO）分期标准,交界性肿瘤组Ⅰ期48例、Ⅱ期3例、Ⅲ期3例;卵巢癌组Ⅰ期6例、Ⅱ期8例、Ⅲ期26例、Ⅳ期5例。③病理类型：交界性肿瘤组以黏液型为主[占56％（30／54）],其次为浆液型[其中普通型11例,微乳头型5例;占30％（16／54）];卵巢癌组以浆液型（其中低度恶性19例,高度恶性3例）为主[占49％（22／45）]。④病理分化程度：卵巢癌组高分化5例,中分化17例,低分化或未分化23例。⑤预后：交界性肿瘤组5年生存率为98％,卵巢癌组为51％,两组比较,差异有统计学意义（P=0．000）。（2）cyclin D1和p53蛋白的表达及其与卵巢癌和交界性肿瘤临床病理特征的相关性：卵巢癌组cyclin D1和p53蛋白的阳性表达率分别为31％（14／45）和56％（25／45）,p53蛋白表达强度与病理分化程度呈正相关（r=0．320,P=0．032）;交界性肿瘤组cyclin D1和p53蛋白的阳性表达率分别为69％（37／54）和6％（3／54）。其中,普通型浆液性交界性肿瘤与高度恶性浆液性癌比较（两者cyclin D1蛋白阳性表达率分别为91％和26％,p53蛋白分别为0和58％）,差异有统计学意义（P〈O．01）;而微乳头型浆液性交界性肿瘤与低度恶性浆液癌比较（两者cyclin D1蛋白阳性表达率分别为3／5和2／3,p53蛋白分别为1／5和1／3）,差异则无统计学意义（P〉0．05）。结论cyclin D1蛋白的过度表达常见于卵巢浆液性交界性肿瘤及低度恶性浆液性癌组织中,而p53蛋白的过度表达更多见于高度恶性浆液性癌组织中。卵巢浆液性交界性肿瘤与高度恶性浆液性癌具有不同的发病机制,而微乳头型浆液性交界性肿瘤与低度恶性浆液性癌的关系可能更为密切。     

子宫颈癌微卫星不稳定性和HPV感染的研究

目的　探讨微卫星不稳定性 (microsatelliteinstability ,MI)及人乳头瘤病毒 (HPV)感染与宫颈癌的相关性。方法　 2 2例宫颈浸润性鳞癌石蜡标本 ,选取 3、 5、 6号染色体上的 3个微卫星位点D3S1 2 89、D5S4 0 6、D6S2 77进行MI分析 ;选用HPV 1 6 / 1 8型特异引物进行HPV检测 ;应用免疫组化法检测Ki6 7蛋白的表达。结果　在宫颈癌标本中 3个位点均未发现MI和杂合性缺失 (LOH)的改变 ;HPV1 6 / 1 8检出率为 77 3% ,Ki6 7指数明显高于对照组。结论　 3个微卫星位点未见MI和LOH的改变 ,与国外报道不同 ,可能与种族差异有关。HPV分型及Ki6 7蛋白表达的检测有助于宫颈病变的评价     

FHIT基因在卵巢癌、子宫内膜癌及宫颈癌细胞系中的表达

3号染色体短臂是卵巢癌、子宫内膜癌及宫颈癌杂合性丢失和细胞遗传学异常的高发区。该区域中存在着著名的“FRA3B脆性位点和t(3;8)断裂位点。”最近,一个新的候选肿瘤抑制基因——脆性组氨酸三联(fragile histdine triad,FHIT)基因被克隆、定位于3p14.2区位。它是脆性组氨酸三联基因家族的成员之一,该基因的总基因组DNA约有1Mb,包含10个外显子,其cDNA长1.1kb。     

外阴鳞状细胞癌组织中染色体变异与人乳头状瘤病毒感染状态的关系

目的 探讨外阴鳞状细胞癌（鳞癌）组织中染色体变异与人乳头状瘤病毒（HPV）感染状态的关系.方法 应用比较基因组杂交（CGH）技术和PCR技术分别检测21例外阴鳞癌患者癌组织中染色体的变异情况和HPV感染状态,并分析两者间的关系.结果 每例患者的外阴鳞癌组织中染色体均有不同程度的变异,包括染色体全部或部分的扩增或丢失,每例患者平均有4.9处（102/21）,其中丢失多于扩增,分别为2.6处（54/21）和2.3处（48/21）.21例患者中,根据染色体变异部位的不同,染色体扩增率依次为3q占43%（9/21）、8q占38%（8/21）、12q占33%（7/21）和9p占19%（4/21）,染色体丢失率依次为4p占52%（11/21）、3p占43%（9/21）和9p占10%（2/21）.在染色体3q及12q部位,HPV阳性的外阴鳞癌组织染色体扩增率（分别为73%、64%）均明显高于HPV阴性者（分别为10%、0,P＜0.01）;而在染色体8q部位,HPV阴性的外阴鳞癌组织染色体扩增率（70%）明显高于HPV阳性者（9%,P＜0.01）.结论 外阴鳞癌组织中存在染色体的扩增或丢失.染色体3q、12q的扩增与外阴鳞癌患者感染HPV有关.     

人宫颈鳞癌和宫颈上皮内瘤变组织内HPV16 DNA及p53基因变异的研究

目的 本文采用常规PCR技术对40例宫颈鳞癌组织学标本和200例宫颈上皮内瘤变（CIN）Ⅰ～Ⅱ级的宫颈分泌物标本的HPV16 DNA及P53基因5～6，7～8外显子变异进行了研究，方法 采用常规PCR技术检测HPV16 DNA和p53基因及免疫组织化学方法检测P53基因。结果 在40例宫颈鳞癌标本中HPV16 DNA阳性检出率有20例（50％）；200例CIN标本中有76例（38％）。P53基因在40例宫颈鳞癌标本中有20例出现变异（50％），其中外显子5缺失10例（50％），外显子6缺失1例（5％），外显子7缺失3例（15％），外显子8缺失6例（30％）。200例CIN标本均未检出p53外显子5～6，7～8的变异，宫颈鳞癌HPVl6DNA阳性与p53基因变异的符合率为80％。结论HPVl6I）NA与宫颈鳞癌的发生有密切的相关性，p53基因在宫颈鳞癌组织中确实存在着变异。本研究认为常规PCR技术作为辅助诊断方法是一种快速、简单、适用于临床的方法。     

卵巢癌患者血清中TPS、CA125水平及其临床意义

目的 探讨卵巢癌患者血清TPS、CA125水平的临床应用价值。方法 分别检测15例卵巢良性肿瘤患者（良性对照组）和39例卵巢癌患者（卵巢癌组）初诊、治疗后以及15例卵巢癌复发患者血清TPS、CA125水平，统计结果进行比较。结果卵巢癌组血清TPS、CA125水平显著高于良性对照组（P〈0．01）；TPS对卵巢癌诊断的灵敏度和特异度为71．8％和86，7％，CA125为84．6％和86．7％，两者差异无显著性（P〉0．05）；治疗后患者血清TPS、CA125水平低于治疗前（P〈0．01、P〈0．05），而复发时TPS水平又显著高于治疗后（P〈0．05）。结论 血清TPS检测有助于卵巢癌诊断、疗效观察及预后判断，联合检测CA125可提高其临床应用价值。     

卵巢上皮性癌相关抗原的筛选和血清学检测

目的构建卵巢上皮性癌（卵巢癌）腹水肿瘤细胞的cDNA文库,从中筛选能用于卵巢癌早期诊断和免疫治疗靶点的卵巢癌相关抗原。方法用3例卵巢浆液性囊腺癌、1例卵巢黏液性囊腺癌、1例卵巢子宫内膜样癌患者的腹水肿瘤细胞构建cDNA文库,采用改良的重组克隆表达抗原的血清学鉴定（SEREX）技术与抑制性消减杂交（SSH）方法相结合的策略从文库中筛选卵巢癌相关抗原基因,并对其进行酶切鉴定、核苷酸测序分析。采用重组肿瘤抗原的微型血清学检测（SMARTA）法检测筛选出的卵巢癌相关抗原与96例卵巢癌患者和96例正常妇女的血清中相应自身抗体的阳性反应情况。结果经两轮血清学筛选和核苷酸测序分析,最终得到55个候选的卵巢癌相关抗原基因片段,代表45个基因,分为6类：（1）与已知的卵巢癌相关基因同源的基因,如BARD1基因等;（2）与其他肿瘤相关基因同源的基因,如TM4SF1基因等;（3）在一些特殊组织中表达的基因,如ILF3、FXR1基因等;（4）与一些特殊功能蛋白基因同源的基因,如TIZ、C1D基因等;（5）与胚胎来源的基因同源的基因,如PKHD1基因等;（6）其余为在基因库GenBank中无同源序列可比对的未知基因,如OV-189基因等。TM4SF1、C1D、BARD1、FXR1、OV-189基因的噬菌体重组融合抗原与卵巢癌患者血清中相应IgG型自身抗体反应的阳性率分别为28％、21％、23％、23％、31％,与正常妇女血清反应的阳性率分别为9％、6％、5％、8％、13％,分别比较,差异均有统计学意义（P〈0．01）;TIZ、FXR1、OV-189基因的重组抗原与卵巢癌患者血清中相应IgM型自身抗体反应的阳性率分别为26％、28％、18％,与正常妇女血清反应的阳性率分别为8％、11％、7％,分别比较,差异均有统计学意义（P〈0．05）。FXR1、OV-189基因的重组抗原与Ⅰ～Ⅱ期卵巢癌患者血清中相应IgG型自身抗体反应的阳性率（分别为34％、46％）高于Ⅲ～Ⅳ期者（分别为16％、23％）,两者分别比较,差异均有统计学意义（P值分别为0．045、0．021）;OV-189基因的重组抗原与高分化卵巢癌患者血清中相应IgG型自身抗体反应的阳性率（为67％）高于中～低分化者（为26％）,两者比较,差异均有统计学意义（P=0．001）。TIZ、FXR1基因的重组抗原与Ⅰ～Ⅱ期卵巢癌患者血清中相应IgM型自身抗体反应的阳性率（分别为40％、46％）高于Ⅲ～Ⅳ期者（分别为18％、18％）,两者分别比较,差异均有统计学意义（P值分别为0．018、0．004）。当联合分析TM4SF1、C1D、TIZ、FXR1基因的重组抗原的相应IgG型自身抗体及TIZ、FXR1基因的重组抗原的相应IgM型自身抗体（即自身抗体谱）时,诊断卵巢癌的敏感度为66％,准确度为73％;将该自身抗体谱与CA125联合时,敏感度、准确度均有明显提高,分别为83％、80％。结论SEREX技术与SSH方法相结合筛选肿瘤抗原基因是一种行之有效的、能筛选出具有较高特异性肿瘤抗原基因的研究策略;TM4SF1、C1D、TIZ、BARD1、FXR1、OV-189基因的重组抗原在血清中的相应自身抗体可作为卵巢癌诊断的标志物,多个抗原的联合检测可提高诊断效率。     

宫颈癌杂合性丢失及新抑癌基因研究进展

肿瘤细胞中存在的染色体区杂合性丢失（LOH）预示着新抑癌基因存在的可能。宫颈癌中存在3p14，3p21-22，3p25，6p21-23和11q22-q24等多个高度LOH区，并在部分区域中鉴定出宫颈癌相关新抑癌基因．如FHIT，RASSF1A和TSLC1等。但因单纯依赖基于微卫星标记的LOH分析和基因突变分析，忽略了染色体纯合缺失及基因启动子区高甲基化所引起的抑癌基因失活，因而鉴定的新抑癌基因甚少。目前LOH、突变分析、DNA甲基化检测、CGH等多种技术的综合应用将为宫颈癌相关新抑癌基因的鉴定带来更多希望。     

应用尿半胱氨酸蛋白酶活性测定诊断妇科恶性肿瘤的价值

对恶性肿瘤７０例（恶性组）、良性肿瘤５０例（良性组）和正常健康妇女５０例（对照组）尿中半抗氨酸蛋白酶（ＵＣＰ）活性进行测定。结果：ＵＣＰ辅助诊断恶性肿瘤的敏感性为９０％、特异性为８０％、准确性为８６％。恶性组ＵＣＰ值明显高于良性组和对照组（Ｐ＜０．０１）。ＵＣＰ检测对卵巢癌尤为敏感，敏感性为９５％，高于子宫体癌（７７％）、宫颈癌（８５％）。ＵＣＰ、血清肿瘤标记物ＣＡ＿（１２５）及乳酸脱氢酶（ＬＤＨ）同功酶辅助诊断卵巢癌的敏感性依次为９０％、８５％、７５％；特异性为８０％、８７％、８５％；准确性为８４％、８４％、８０％。ＵＣＰ与ＣＡ＿（１２５）对卵巢癌的辅助诊断有同样的价值，而且优于ＬＤＨ同功酶。８例卵巢癌Ⅰ期患者，７例ＵＣＰ值异常，而同组６例卵巢癌Ⅰ期患者中，无１例ＣＡ＿（１２５）值异常，说明ＵＣＰ检测对早期卵巢癌较ＣＡ＿（１２５）更敏感。提示：ＵＣＰ可能成为妇科恶性肿瘤，特别是卵巢癌的标记物。     

Loss of heterozygosity on chromosome 17q in epithelial ovarian tumors: association with carcinomas with serous differentiation.

Loss of heterozygosity (LOH) on chromosome 17q is frequent in epithelial ovarian tumors, but its clinicopathologic significance remains to be elucidated. DNA of 50 patients with epithelial ovarian tumors was extracted from blood and from fresh-frozen and paraffin-embedded tissue (14 benign, 7 borderline, and 29 malignant). Six microsatellite loci were amplified by PCR (D17S250, TRHA1, D17S800, D17S855, D17S579, D17S513). LOH was scored by the absence or reduction of the signal to less than 50% of one of the alleles in tumor DNA compared with normal DNA. LOH was identified on chromosome 17q in at least one locus in 12 tumors (24%), all of them carcinomas (12 of 29 tumors, 41.3%). It occurred more frequently among high-grade serous carcinomas (8 of 14 tumors, 57%) and mixed endometrioid-serous carcinomas (2 of 5, 40%). LOH was detected in all informative markers of 10 tumors, suggesting the complete loss of an entire chromosome 17 homologue. Patients with LOH-positive carcinomas were older than those with LOH-negative malignant tumors (mean ages 67 and 49). The results support the hypothesis that LOH on chromosome 17q may be associated with the development of ovarian cancers in elderly patients, particularly with high-grade serous or mixed endometrioid-serous carcinomas.     

Microsatellite analysis in serous tumors of the ovary.

Thirty-four serous ovarian neoplasms were analyzed for microsatellite instability (MIN) and loss of heterozygosity (LOH) at 20 chromosomal loci of eight different chromosomes, including the map positions of the six known mismatch repair genes. Twelve of the 20 (60%) serous ovarian tumors of low malignant potential (LMP) and 13 of 15 (87%) samples of serous ovarian carcinomas, taken from 14 patients, showed LOH at one or more of the analyzed microsatellite loci. In serous carcinomas, LOH of the dinucleotide repeat D7S521 was most frequent. Six of 20 (30%) LMP tumors were also affected by MIN at more than one locus, whereas in the carcinomas MIN was found only sporadically (p < 0.025). No correlation between increased occurrence of MIN and LOH at the chromosomal loci of the known mismatch repair genes were discovered for these LMP tumors. Although our observation regarding LOH frequency in serous LMP tumors and serous carcinomas is compatible with the hypothesis that serous LMP tumors might represent precursor lesions of invasive carcinomas, the results concerning the occurrence of MIN suggest that the mechanisms of tumorigenesis of some tumors of LMP are distinct from those in invasive serous carcinomas.     

Allelic deletion mapping on chromosome 6q and X chromosome inactivation clonality patterns in cervical intraepithelial neoplasia and invasive carcinoma

OBJECTIVE: Loss of heterozygosity (LOH) profiles and X chromosome inactivation patterns are analyzed in 42 patients with cervical intraepithelial neoplasias (CIN), including low-grade (CIN1) and high-grade (CIN2, CIN3) lesions, and 22 patients with invasive cervical carcinomas. METHOD: Laser capture microdissection was utilized to procure pure matched normal and lesional cells from each case. Sixteen microsatellite markers on four chromosomal arms, 6q21-q25.1, 8p21, 13q12.3--q13, and 17q12--q21, were amplified for LOH, as well as the HUMARA locus for X chromosome inactivation analysis. Eight additional markers spanning the long arm of chromosome 6 were utilized in all cases showing LOH on this arm and in which further tissue material was available for microdissection. RESULTS: Fifty-five percent of carcinomas showed deletions on chromosome bands 6q21--q25.1, 43% on 13q12.3--q13, and 40% on 17q12--q21. Deletions on 6q were identified in CIN3 (40%), CIN2 (37%), and CIN1 (10%), on 13q in CIN3 (33%) and CIN2 (33%), and rarely on chromosomal arm 17q. Finer 6q mapping revealed that marker D6S310 (q22) represented the centromeric and marker D6S255 (q25--q16) the telomeric boundary of deletion. A second, telomeric area of deletion at marker D6S281 (q27) was also identified. Monoclonal X chromosome inactivation patterns were identified in 12/13 cancers, 13/14 CIN3, 5/10 CIN2, and 0/6 CIN1. CONCLUSIONS: Two areas of deletion on chromosome 6q were identified in cervical tumors, suggesting the presence of tumor suppressor gene(s) inactivated in this neoplasia. LOH on this arm were identified early during cervical tumor progression. LOH on 13q and 17q also occur in cervical cancers. X chromosome inactivation patterns suggest that CIN develops into a monoclonal lesion during progression from CIN1 to CIN3.     

Xp22.2-3 loss of heterozygosity is associated with germline BRCA1 mutation in ovarian cancer

OBJECTIVE: X-Chromosome loss of heterozygosity (LOH) occurs in approximately 40% of ovarian cancers. We have previously demonstrated an association between nonrandom X-chromosome inactivation and germline BRCA1 mutation. The current study examines the association between X-chromosome LOH and BRCA1 mutation. METHODS: Ninety tumor DNA (81 ovary, 5 fallopian tube, 4 primary peritoneal) and matched peripheral blood mononuclear cell DNA samples were examined for LOH with 11 X-chromosome microsatellite DNA markers. RESULTS: Tumor DNA demonstrated frequent LOH at the Xp22.2-3 region (37.7% at DXS6807). Loss of heterozygosity on Xp was twice as common in tumor DNA from germline BRCA1 mutation carriers (9/14 vs 19/67, P = 0.02). In four evaluable samples, Xp22.2-3 LOH preferentially occurred from the active X allele. CONCLUSIONS: Our data support the hypothesis that an Xp22.2-3 gene product interacts with or modifies the expression of BRCA1 in some hereditary ovarian cancers.     

Investigation of loss of heterozygosity at specific loci on chromosomes 3p, 6q, 11p, 17p and 17q in ovarian cancer

In an attempt to further define the genetic events in the pathogenesis and progression of human ovarian cancer, an analysis of constitutional and ovarian carcinoma DNA samples revealed loss of heterozygosity (LOH) at specific loci on chromosomes 3p (38%), 6q (23%), 11p (33%), 17p (82%) and 17q (62%). In contrast, LOH was not observed in benign or borderline tumors. No significant association could be demonstrated between LOH at the loci studied and tumor stage, histologic subtype, grade or patient outcome. Further analyses of large tumor panels are now required to determine the relationship between LOH at these loci and the clinicopathological behavior of ovarian tumors.     

Patterns of loss of heterozygosity at 10q23.3 and microsatellite instability in endometriosis, atypical endometriosis, and ovarian carcinoma arising in association with endometriosis.

Genetic aberrations, such as loss of heterozygosity (LOH) and mutations leading to functional inactivation of the PTEN tumor suppressor gene, located on chromosome 10q23.3, have been shown to be associated with approximately one third of ovarian adenocarcinomas. In addition, microsatellite instability (MSI) leading to the functional inactivation of the PTEN gene has also been reported for ovarian adenocarcinomas with frequencies varying from 6 to 37%. However, the frequency of PTEN gene abnormalities has not been well studied or evaluated in lesions such as typical and atypical endometriosis. The aim of this study was to investigate a possible sequential progression from endometriosis through atypical endometriosis to ovarian carcinoma by assessing LOH at 10q23.3 and MSI in those entities. Genomic DNA was analyzed for LOH and MSI at 3 loci on chromosome 10, using polymerase chain reaction amplification. Significant differences in LOH were seen between endometriosis (4.3%) and ovarian carcinoma (23.5%) at D10S608. The differences at the other 2 loci were not significant. A high frequency of MSI was found in endometriosis (82.6%) and atypical endometriosis (75%); however, the differences were not significant. These results suggest that LOH at D105608 may possibly be an important molecular event in the progression of endometriosis to carcinoma. This study highlights that endometriosis and atypical endometriosis might act as precursor lesions that have the potential to progress into ovarian adenocarcinoma.     

Allele loss on chromosome 1p36 in epithelial ovarian cancers.

OBJECTIVES: Prior studies have shown that allelic loss on chromosome 1p36 occurs frequently in ovarian as well as several other types of cancer. This suggests that inactivation of gene(s) in this region may play a role in the pathogenesis of these cancers. The aim of this study was to further delineate the region of loss on chromosome 1p36 in ovarian cancers and to identify associated patient or tumor characteristics. METHODS: Paired normal/cancer DNA samples from 75 ovarian cancers (21 early stage I/II and 54 advanced stage III/IV) were analyzed using microsatellite markers. RESULTS: Forty-nine of 75 (65%) ovarian cancers had loss of at least one marker. The marker demonstrating the most frequent loss was D1S1597, which was lost in 29/57 (51%) informative cases. Allele loss on 1p36 was significantly more common in poorly differentiated ovarian cancers (73%) relative to well or moderately differentiated cases (48%) (P = 0.03). Evidence was obtained for two common regions of deletion: one flanked by D1S1646/D1S244 and another more proximally by D1S244/D1S228. CONCLUSION: These findings further delineate regions on chromosome 1p36 proposed to contain tumor suppressor gene(s) that may play a role in the development and/or progression of epithelial ovarian carcinoma. Allele loss on 1p36 is associated with poor histologic grade.     

Detecting and investigating the significance of high-frequency LOH chromosome regions for endometriosis-related candidate genes

To detect the high-frequency loss of heterozygosity (LOH) chromosome regions for ectopic endometrium of ovarian endometriosis (EMs) and to investigate the significance of high-frequency LOH chromosome regions in EMs, we obtained ectopic endometrium by laser capture microdissection (LCM (22 samples)), manual capture microdissection (MCM (18 samples)), and routine dissection (14 samples), respectively. After restriction and circularization-aided rolling circle amplification (RCA-RCA), LOH was detected at 12 microsatellite (MS) loci. The frequency of LOH was 59.09% (13/22) in LCM group, 61.11% (11/18) in the MCM group and 21.43% (3/14) in the routine dissection group. The latter was significantly lower when compared with the former two (p < 0.05). In the LCM group, candidate chromosome regions 17q21.31 and 9p21.3 had LOH frequencies of 23.8 and 13.6%, respectively. The highest LOH frequency was detected at the locus AAAT2 on chromosome 17q21.31 (40%). The chromosome region with the highest frequency of LOH for ectopic endometrium was 17q21.31, especially at the AAAT2 locus, which prompted that down regulation of the candidate genes nearby the locus might be one of the mechanisms of EMs pathogenesis. LCM combined with RCA-RCA is a reliable technique for analyzing endometrial LOH at multiple MS loci. MCM combined with RCA-RCA, which provided similar results, was more cost-effective.