摘要
背景：子宫内膜异位症(EM)是妇科的常见疾病,是指子宫内膜组织(包括腺体和间质)在子宫内膜以外的部位生长。EM的病因尚不明确,目前的诸多学说均不能完全解释临床上多种类型内异症的发病。因此,进一步探究内异症的病因对于深入理解该病的发病机制、寻找更准确的诊断方法以及探索更有效的治疗途径,都有十分重要的意义。随着对EM临床基础研究的不断深入,子宫内膜异位症是一种干细胞疾病的假说被提出,并逐渐成为内异症发病机制研究的新方向和新热点。
目的：本课题拟通过检测X-连锁的人类雄激素受体(HumAR)基因的杂合缺失情况,验证EM病灶是否具有干细胞的单克隆特性,以及不同类型EM病灶的克隆性是否存在差异,从单克隆性的角度验证内异症干细胞起源假说。
方法：本研究对象为各种类型EM病灶的新鲜组织标本,利用聚合酶链反应(PCR)和基因片段分析技术,对EM病灶组织限制性核酸内切酶Hhal酶切前后的HumAR基因进行扩增和分析,检测该基因的杂合缺失情况,验证组织的克隆性。并对部分内异症病灶组织进行显微切割,检测单个腺体以及间质的克隆性,分析不同腺体之间、腺体与间质之间的克隆性是否存在差异。
结果：大部分子宫内膜异位症整个病灶均来源于单个克隆(55.1%),不同类型EM的单克隆比例有所差异,其中巧囊为44.9%(31例/69例),盆腔腹膜EM为84.6%(11例/13例),DIE为100%(6例/6例),腹壁EM为64.3%(10例/14例),会阴EM为40%(2例/5例)。巧囊、盆腔腹膜EM、DIE、腹壁EM与自体对照(子宫内膜/正常卵巢上皮／正常腹壁会阴组织)相比,单克隆比例明显增高(P<0.05)。会阴EM与自体对照相比,单克隆比例无显著差异(P>0.05)。同一患者不同EM病灶各自表现单克隆特性,但可以来源于不同克隆。对多克隆的巧囊和会阴EM组织进行显微切割,发现单个EM腺体为单克隆,但不同腺体可以来源于不同克隆。
结论：大多数EM病灶表现为单克隆,某些特殊类型EM(如DIE)具有显著的单克隆性；同一患者不同部位EM病灶可以各自呈现单克隆特性；多克隆EM组织其单个腺体亦来源于单一克隆。证明子宫内膜异位症病灶具有单克隆特性,但同一患者不同病灶,或同一病灶不同的腺体可以来源于不同克隆。我们的研究提示EM具有干细胞的主要特征之一,即单克隆性。关于EM干细胞的其它特性和EM病灶的干细胞来源,尚待更深入的探索。
BACKGROUND:Endometriosis is a common gynecological disorder. It is defined by the presence of endometrial tissue outside the uterine cavity. The pathogenesis of endometriosis remains unclear. The current etiological theories can not folly explain clinical varieties of endometriosis. It is important to explore the pathogenesis of endometriosis and look for new methods of treatment. A series of recent research have suggested that dislocation of basal endometrial stem cell might result in endometriosis and it might be a stem cell related disorder.
OBJECTIVE:The proliferation clonality of endometriosis lesions is detected with a method of molecular analysis. The stem cell hypothesis of EM will be tested.
METHODS:The heterozygosity of HumAR gene of obtained endometriosis lesions were identified by a polymerase chain reaction (PCR) assay and STR analysis. The clonality of EM gland and stromal were detected after microdissection.
RESULTS:55.1%of EM lesions were monoclonal. The monoclonal proportions of different EM lesions were different:OEM44.9%(31/69), PEM84.6%(11/13), DIE100%(6/6), Abdominal EM64.3%(10/14), Perineum EM40%(2/5). Compared with endometrial tissue and normal tissue, the monoclonal proportion was significantly increased in OEM/PEM/DIE/Abdominal EM (P<0.05), not significantly increased in Perineum EM (P<0.05). Polyclonal EM lesions were microdissected and every single gland was monoclonal, but different glands could be from different monoclonal origins.
CONCLUSIONS:Most EM lesions are monoclonal. Certain types of EM (e.g.DIE) show the monoclonality significantly. Different EM lesions from the same patient can have different monoclonal origins. A single endometiosis gland is monoclonal. It revealed that endometiosis could derive from a single stem cell, but different glands/lesions could be from different stem cells. This research showed a certain relationship between endometriosis and stem cells. But more basic research and clinical evidence will be needed to support the stem cell hypothesis of EM.
引文
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