人类雄激素受体基因检测子宫内膜异位症病灶克隆性

Clonal analysis of endometriotic lesions using human androgen receptor gene

目的通过检测X染色体连锁的人类雄激素受体基因（HUMARA）多态性，研究各种病理类型子宫内膜异位症（内异症）病灶的克隆性特征。方法收集2008年12月—2009年6月在北京协和医院因内异症接受腹腔镜或开腹手术治疗的6例患者的8份冰冻组织标本，利用激光捕获显微切割（LCM）技术在异位内膜中采集50个单个腺体的上皮细胞，以HUMARA作为基因标志物，采用巢式PCR技术、HhaI和HpaⅡ双酶切反应及全自动激光荧光测序进行等位基因片段长度多态性检测，以此判定组织的克隆性特征。结果50个内异症腺体中，34个腺体标本符合克隆性分析要求，均为单克隆性。3例卵巢内异症病灶标本中，有2例符合克隆性分析要求，1例标本的4个腺体克隆性模式一致，另1例的7个腺体克隆性模式一致。2例盆腔内异症病灶标本中，1例仅有1个腺体可供克隆性分析，另1例的5个腺体克隆性模式不一致。2例腹壁内异症病灶标本中，1例的7个腺体克隆性模式不一致；另1例的6个腺体克隆性模式不一致。1例深部浸润型内异症标本的4个腺体克隆性模式一致。同一患者不同病理类型的内异症病灶腺体克隆模式均不相同。结论内异症病灶中的单个腺体为单克隆，但同一病灶中的不同腺体克隆模式可能不尽相同。

Objective To investigate characteristics of endometriotic lesions clone with various pathological subtypes by analyzing the polymorphism of X chromosome linked human androgen receptor allele （HUMARA）. Methods Eight frozen tissues of endometriotie leisons were collected from 6 patients who received laparotomy or laparoscopy surgery in Peking Union Medical College Hospital from Nov. 2008 to Jun. 2009. Fifty specimens of epithelial cells from single endometrial glands were isolated and collected from endometriotic lesions by using laser capture microdissection. HUMARA was applied as the gene marker of clonal analysis. Nested polymerase chain reaction, double-enzyme digestion reaction with two methylationsensitive restriction endonuclease （Hha Ⅰ and Hpa Ⅱ ） , and the automated gene sequencing technique were utilized in this study to evaluate the characteristics of endometriotic lesions clone. Results Of 50 specimens of isolated glands, 34 were informative for clonal analysis, and all of which showed monoclonality. Of 3 ovarian endometriotic tissues, one tissue of HUMARA showed unuseful information, the other 2 ovarian endometriotie tissues respectively had 4 and 7 informative specimens of gland epithelial cells, and all of the glands from each tissue showed uniform clonal pattern. Two peritoneal endometriotic tissues had 1 and 5 informative specimens from individual glands, respectively; and the clonal patterns in 5 glands from the single lesion were divergent. Two abdominal wall endometriotic tissues had 7 and 6 informative specimens, respectively; and variable clonal patterns were seen in different glands from each lesion. One deep infiltrating endometriotic lesion had 4 informative specimens of isolated glands, and all of them showed unique clonal pattern. The disparate clonal patterns were found in endometriotic lesions with variable pathological subtypes, even arising from the same patient. Conclusions The epithelial cells from individual endometriotic gland showed monoclonality , and different glands from the same endometriotie lesion might show divergent patterns.