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Use for research materials in Pathology--application for molecular pathology

病理学研究材料的用途--分子病理学的应用

基本信息

批准号：
63480143
负责人：
MAEDA Sakan
金额：
$ 0.38万
依托单位：
Kobe University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1988
资助国家：
日本
起止时间：
1988 至 1989
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63480143/
关键词：
Formalin fixation Molecular pathology Oncogenes DNA degradation Gene expression Culture Monoclonal antibody

项目摘要

Department of Pathology has many chance to get materials from surgical operation and autopsy. However, their uses may be limited to histological diagnosis and other trials to use for another purpose must be necessary. Present experiment was undertaken to examine how to use l.fresh, 2.unfixed, and 3.fixed pathological materials for pathological research. In 1., the culture supernatant of human placenta contained the growth factor for human squamous cell lines and it further was examined which factor might be essential for that stimulation. For lung small cell carcinomas, L-cysteine, transferrin, and bathocuproine disulfo hate had also growth activity and L-cysteine was most important. In 2., extraction method for RNA was examined and modification of Rupp and Locker's method might be effective for massive extraction from degenerated tissues. In 3., the mechanism of degeneration of formalin-fixed tissues was examined basically. The extracted and purified DNA was not damaged when it was fixed in 10% formalin.Because pulsed field gel electrophoresis and histone-DNA recombination experiment revealed the same damage to formalin-fixed tissues when it was extracted by usual DNA extraction method, the mechanical damage was not so important as the mechanism of degradation. Fixation in low temperature and addition of EDTA inhibited the degradation of DNA. Enzymatic activity such as DNase during formalin fixation might have responsibility for DNA degradation. Monoclonal antibodies which were specific to germ cells and germ cell tumors, were generated from formalin-fixed and parrafin-embedded tissues as mouse immunogen.Several trials might become possible to use many pathological specimens for pathological research. However further experiments may be necessary for multi- purpose use of them.

病理科有很多机会从外科手术和尸检中获得材料。然而，它们的用途可能仅限于组织学诊断，必须有必要进行其他试验以用于其他目的。本实验探讨了如何利用新鲜、未固定和固定的病理材料进行病理学研究。在1.，人胎盘的培养上清液含有人鳞状细胞系的生长因子，并进一步检测哪种因子可能是刺激所必需的。对于肺小细胞癌，L-半胱氨酸、转铁蛋白和浴铜灵二磺酸盐也具有生长活性，并且L-半胱氨酸是最重要的。在2.，本文对RNA的提取方法进行了研究，并对Rupp和Rupp的方法进行了改进，可有效地从变性组织中大量提取RNA。在3.，对福尔马林固定的组织的变性机理进行了基本的检查。提取纯化的DNA经10%福尔马林固定后无损伤，但脉冲场凝胶电泳和组蛋白-DNA重组实验表明，常规DNA提取方法对福尔马林固定的组织也有损伤，因此机械损伤不如降解机制重要。低温固定和EDTA的加入抑制了DNA的降解。福尔马林固定过程中的酶活性（如DNA酶）可能是DNA降解的原因。用福尔马林固定石蜡包埋的组织作为小鼠免疫原，制备了生殖细胞和生殖细胞肿瘤特异性单克隆抗体，并进行了多种病理标本的病理学研究。然而，进一步的实验可能是必要的多用途使用它们.