Iwamoto K S, Mizuno T, Ito T, Akiyama M, Takeichi N, Mabuchi K, Seyama T
Department of Radiobiology, Hiroshima University School of Medicine, Japan.
Am J Pathol. 1996 Aug;149(2):399-406.
Archival tissues are a bountiful resource for various studies. Polymerase chain reaction permits the use of such tissues for molecular biological analyses of disease causation. However, a comprehensive study using a large number of decades-old samples (20 or more years) for molecular oncology/epidemiology has never been shown to be feasible. We have relied upon the unique tumor registry of atomic bomb survivors to show that such studies are possible using 275 hepatocellular carcinoma and 41 skin cancer cases. We used 23 relatively recent thyroid papillary carcinoma cases from persons living in the vicinity of the Chernobyl nuclear reactor accident for comparison. Degradation of DNA is severe in autopsy hepatocellular carcinoma samples but can be compensated for by decreasing the polymerase chain reaction product size. Increasing the amount of DNA that is used by a factor of 8 improved amplification efficiency from approximately 60 to 80%. Age of the samples was not as great a problem as was the source of procurement. The extracted DNA can be used for all types of assays that require polymerase chain reaction amplification, such as restriction fragment length polymorphism, single-strand conformation polymorphism, and direct sequencing.
存档组织是各类研究的丰富资源。聚合酶链反应使得能够利用此类组织对疾病病因进行分子生物学分析。然而,使用大量数十年前(20年或更久)的样本进行分子肿瘤学/流行病学的全面研究从未被证明是可行的。我们借助原子弹幸存者独特的肿瘤登记资料表明,利用275例肝细胞癌和41例皮肤癌病例进行此类研究是可行的。我们使用了23例来自切尔诺贝利核反应堆事故附近居民的相对近期的甲状腺乳头状癌病例作为对照。尸检肝细胞癌样本中的DNA降解严重,但可通过减小聚合酶链反应产物大小来弥补。将所使用的DNA量增加8倍可使扩增效率从约60%提高到80%。样本的年龄不像取材来源那样是个大问题。提取的DNA可用于所有需要聚合酶链反应扩增的检测,如限制性片段长度多态性、单链构象多态性和直接测序。