|Budget Amount *help
¥2,400,000 (Direct Cost : ¥2,400,000)
Fiscal Year 1996 : ¥1,000,000 (Direct Cost : ¥1,000,000)
Fiscal Year 1995 : ¥1,400,000 (Direct Cost : ¥1,400,000)
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
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.
今回の研究を進めて行くうちに、differental display法によるcDNAクローニングにいくつかの難題が見いだされたため、子宮内膜症に特有の遺伝子変異を明確にするために細胞生物学的な解析も行った。2方向からその解析を行い、1)子宮内膜症患者血清中に検出される抗子宮内膜抗体の特異性と生物活性(Mizuno,Umesaki,Ogita et al.JSRI in press,1997;Mizuno,Umesake,Ogita et al.投稿準備中)、2)子宮内膜症患者と正常婦人から樹立した長期培養正所性子宮内膜組織の細胞学的性状(田中、梅咲、荻田ら,エンドメトリオージス研究会誌,1997;Tanaka,Umesaki,Ogita et al.投稿準備中)、について内膜症特異遺伝子クローニングのための別の足掛りを得た。これらに関しては、現在も解析中にある。 Less