Molecular cloning and clinical application of the endometriosis-specific genes

为了成功完成本研究，以下两个假设必须是可重复的：第一，子宫内膜异位症异位子宫内膜组织与非子宫内膜异位症异位子宫内膜组织的mRNA表达存在显著差异；二是差异显示法克隆cDNA重复性高。然而，我们发现了意想不到的问题，我们不得不改变这项研究的方向。问题如下：虽然发现异位子宫内膜间质细胞在子宫内膜异位症患者和非子宫内膜异位症女性之间的生物活性有一定的差异，但就我们的研究而言，并没有发现它们之间存在稳定的遗传差异。不稳定差异的可能原因表明，临床健康的女性可能有很少的子宫内膜异位症病变，子宫内膜异位症可能引起继发性刺激，影响异位子宫内膜间质细胞的异常基因表达。对于差分显示，我们采用了原方法，按原程序随机使用12m。但是，正如我们之前在本研究的中间报告中描述的那样，这种方法碰巧扩增了少量被污染的cDNA和/或分离出不可复制的cDNA。差分显示法克隆cDNA的成功报道尚不多见，该方法刚问世时被认为是一种令人兴奋的cDNA克隆方法。因此，我们对差分显示进行了一些修改，但都没有成功。由于我们在进行多次实验时发现了本研究中的一些技术问题，下面从子宫内膜异位症的细胞性质来识别和阐明子宫内膜异位症的具体遗传疾病。最近我们发现，长期体外培养的异位子宫内膜间质细胞在子宫内膜异位症患者和非子宫内膜异位症患者中某些表面抗原的表达存在差异，并且在子宫内膜异位症患者血清中发现的抗子宫内膜异位症抗体具有一定的组织特异性和特异性生物活性。这些结果可能为克隆子宫内膜异位症相关基因的cdna提供一些线索。少

To complete this study successfully, the following two hypotheses should have been reproducible : First, distinct difference in mRNA expressions between endometriotic eutopic endometrial tissues and non-endometriotic eutopic endometrial tissues ; Second, high reproducibility in cDNA cloning by the method of differential display. However we found unpredicted problems by which we had to change the direction of this study. The problems were as follows : Although there found certain differences in biological activities of eutopic endometrial stromal cells between endometriotic patients and non-endometriotic women, we could not find any stable genetic differences in them as far as we investigated. Possible causes of the unstable differences indicate that clinically healthy women can have minimal endometriotic lesions in them, and that endometriosis may cause secondary stimuli which can affect abnormal gene expressions in eutopic endometriotic stromal cells. As for differential display, we a … More pplied the original method by using random 12mers according to the original procedure. But this method, as described before in our previous intermediate report on this study, happened to amplify a minor contaminated cDNAs and/or to isolate an unreproducible cDNA.Few successful report in cDNA cloning by differential display has been published yet in fact, this method was thought to be exciting approach of cDNA cloning when the method was published. Accordingly we have tried some modifications in differential display in vain. Since we have found some technical problems in this study when we performed many experiments, the following cellular natures in endometriosis to identify and clarify the specific genetic disorders to endometriosis. Recently we have disclosed that long-term in vitro cultured eutopic endometrial stromal cells show the differential expressions of some surface antigens between endometriotic patients and non-endometriotic women, and that anti-endometriotic antibodies found in endometriotic patient sera have certain tissue-specificity and specific biological activity. These results may provide some clues to clone cDNAs for endometriosis-related genes. Less