Molecular cloning and clinical application of the endometriosis-specific genes

子宫内膜异位症特异性基因的分子克隆及临床应用

• 批准号: 07807156
• 负责人: OGITA Sachio
• 金额: $ 1.54万
• 依托单位: OSAKA CITY UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07807156/
• 关键词: Endometriosis; cDNA cloning; endometrium; differential display; 間質細胞

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

为了成功地完成这项研究，以下两个假设应该是可重复性的：第一，异位内膜在位内膜组织和非内异位在位内膜组织之间的mRNA表达存在明显差异；第二，差异显示法克隆的cDNAs具有较高的重复性。然而，我们发现了一些不可预测的问题，通过这些问题，我们不得不改变了本研究的方向。问题是：虽然我们发现子宫内膜异位症患者和非子宫内膜异位症患者在位内膜基质细胞的生物学活性存在一定的差异，但我们目前还没有发现它们之间有任何稳定的遗传差异。这种不稳定差异的可能原因表明，临床健康女性可能存在微小的子宫内膜异位症病变，而子宫内膜异位症可能会引起继发性刺激，从而影响在位子宫内膜异位间质细胞的异常基因表达。至于差异显示，我们是…并按原程序采用随机12聚体的方法。但这种方法，正如我们之前关于这项研究的中期报告中所描述的那样，恰好扩增了少量污染的cDNA和/或分离出了不可重复性的cDNA。目前，差异显示法克隆cDNA的报道还很少，事实上，当该方法发表时，人们认为这种方法是一种令人兴奋的cDNA克隆方法。因此，我们在差异显示方面尝试了一些修改，但都是徒劳的。由于在本研究中我们在进行多次实验时发现了一些技术问题，下面就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障碍。最近，我们披露了长期体外培养的在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存在某些表面抗原的差异表达，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提供一些线索。较少

项目成果

Chang L,Tanaka T,Mizuno K,Chen H,Miyama M,Umesaki N,Ogita S: "Regulating system in susceptibility to apoptosis in human endometrial epithelium : Regulation by specific binding of endometrial cells to extracellular matrices." JSIR. 11 (in press). (1997)

Chang L，Tanaka T，Mizuno K，Chen H，Miyama M，Umesaki N，Ogita S：“人子宫内膜上皮细胞凋亡敏感性的调节系统：子宫内膜细胞与细胞外基质特异性结合的调节。”

Tanaka T,Mizuno K,Miyama M,Nakamura H,Mine M,Hashiguchi Y,Umesaki N,Ogita S,Yamanoto I,Umeki K,Ohtaki S.: "Cytokine regulation of Fas-mediated apoptotic signals in human endometrial epithelium." JSIR. 10. 21-22 (1996)

Tanaka T，Mizuno K，Miyama M，Nakamura H，Mine M，Hashiguchi Y，Umesaki N，Ogita S，Yamanoto I，Umeki K，Ohtaki S.：“人子宫内膜上皮中 Fas 介导的细胞凋亡信号的细胞因子调节。”

Chang L.et al.: "Regulating system in susceptibility to apoptosis in human endometrial epithelium : Regulation by specific binding of endometrial cells to extracellular matrices" proceeding of 11th JSIR. (in press). (1997)

Chang L.等人：“人子宫内膜上皮细胞凋亡敏感性调节系统：通过子宫内膜细胞与细胞外基质的特异性结合进行调节”第11期JSIR会议论文集。

Tanaka T.et al.: "Establishmcnt and characterization of B1-derived macrophage cell line:Effects of anti-endometriotic medicine,Danazol,on the macrophage cells and the parent B1 cells." Proceeding of 11th JSIR. (in press). (1997)

Tanaka T.et al.：“B1 衍生巨噬细胞系的建立和表征：抗子宫内膜异位药物达那唑对巨噬细胞和亲本 B1 细胞的影响。”

Tanaka T.et al.: "Cytokine regulation of Fas-mediated apoptotic signals in human endometrial epithelium" proceeding of 9th JSIR. 10. 21-22 (1996)

Tanaka T.et al.：“人子宫内膜上皮中 Fas 介导的细胞凋亡信号的细胞因子调节”第 9 届 JSIR 会议论文集。