Molnar Bela, Bocsi Jozsef, Karman Jozsef, Nemeth Annamaria, Pronai Laszlo, Zagoni Tamas, Tulassay Zsolt
Clinical Gastroenterology Unit, Second Department of Medicine, Semmelweis University of Medicine, 1088 Budapest, Hungary.
Anticancer Res. 2003 Jan-Feb;23(1B):655-60.
Quantitative DNA analysis of fresh biopsy material can contribute to a more accurate diagnosis and prognosis. The authors aimed to develop and test a mechanical, nuclear preparation protocol for quantitative DNA analysis.
Altogether 32 gastric (10 healthy, 17 gastritis, 7 adenocarcinoma) and 48 colon (21 healthy, 20 colitis ulcerosa, 7 adenocarcinoma) biopsy specimens were evaluated. The mechanical disruption was performed by Medimachine (DAKO, Denmark). The flow cytometry analysis was performed on a BD FACSStar flow cytometer.
DNA Aneuploidy was found in gastric samples only in tumours. The S-phase fraction of the normal cases was 5.9 +/- 2.1%, 5.1% +/- 1.2% in gastritis and 10.7 +/- 1.6% in carcinomas. Seven out of 20 colitis ulcerosa and 4 out of 7 colon cancer samples were aneuploid. The S-phase fraction of normal colon cases was 5.7 +/- 3.4%, in colitis 8.1 +/- 4.2% and 15.1 +/- 5.7% in carcinomas, respectively.
Mechanical nuclear isolation is a useful method for flow cytometric DNA ploidy analysis of fresh biopsy samples.
对新鲜活检材料进行DNA定量分析有助于更准确地诊断和判断预后。作者旨在开发并测试一种用于DNA定量分析的机械细胞核制备方案。
共评估了32份胃活检标本(10份健康标本、17份胃炎标本、7份腺癌标本)和48份结肠活检标本(21份健康标本、20份溃疡性结肠炎标本、7份腺癌标本)。采用Medimachine(丹麦达科公司)进行机械破碎。使用BD FACSStar流式细胞仪进行流式细胞术分析。
仅在胃肿瘤样本中发现DNA非整倍体。正常病例的S期细胞分数为5.9±2.1%,胃炎病例为5.1%±1.2%,癌病例为10.7±1.6%。20份溃疡性结肠炎样本中有7份、7份结肠癌样本中有4份为非整倍体。正常结肠病例的S期细胞分数分别为5.7±3.4%,结肠炎病例为8.1±4.2%,癌病例为15.1±5.7%。
机械细胞核分离是对新鲜活检样本进行流式细胞术DNA倍体分析的一种有用方法。