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结直肠肿瘤患者正常外观结肠黏膜DNA合成期比例的流式细胞术分析

Flow cytometric analysis of DNA synthetic phase fraction of the normal appearing colonic mucosa in patients with colorectal neoplasms.

作者信息

Nakamura S, Goto J, Kitayama Y, Sheffield J P, Talbot I C

机构信息

Division of Pathology, School of Medicine Iwate Medical University, Japan.

出版信息

Gut. 1995 Sep;37(3):398-401. doi: 10.1136/gut.37.3.398.

Abstract

DNA synthetic (S) phase fractions of normal appearing colonic mucosa in Japanese and British patients with colorectal neoplasms were compared with those in patients without colonic neoplasms. Normal crypts were isolated from fresh surgical specimens of the large intestine by the use of EDTA. After fixation with 70% ethanol, isolated crypts were digested with pepsin into single nuclei suspensions. These were stained with propidium iodide and examined by flow cytometry. S phase fraction was calculated from the flow cytometry DNA histogram using Baisch's method. S phase fractions of normal appearing crypts in Japanese and British patients with colorectal tumours were not significantly different and analysed together. S phase fraction of normal appearing colonic crypts in 14 patients with familial adenomatous polyposis (FAP) was 10.23 (2.59%) (mean SD)) ranging from 5.8 to 18.8. S phase fraction of background normal mucosal in patients with large adenomas (over 2 cm) and adenocarcinomas were 9.74 (3.76%) (range, 2.7-16.1) and 8.93 (3.54%) (range, 2.9-18.9) respectively. In normal mucosa of patients without any colorectal neoplasms, S phase fraction was 8.99 (3.94)% (range, 3.9-17.7). There was no statistically significant difference in S phase fractions of normal mucosa in the four groups. Our results show that an increase in proliferative activity of background colonic crypts is not necessary for tumour development.

摘要

比较了患有结肠直肠肿瘤的日本和英国患者正常外观结肠黏膜的DNA合成(S)期比例与无结肠肿瘤患者的相应比例。通过使用乙二胺四乙酸(EDTA)从大肠新鲜手术标本中分离出正常隐窝。用70%乙醇固定后,将分离出的隐窝用胃蛋白酶消化成单核悬液。这些悬液用碘化丙啶染色并通过流式细胞术进行检测。使用拜施方法从流式细胞术DNA直方图计算S期比例。将患有结肠直肠肿瘤的日本和英国患者正常外观隐窝的S期比例合并分析,二者无显著差异。14例家族性腺瘤性息肉病(FAP)患者正常外观结肠隐窝的S期比例为10.23(标准差2.59%),范围为5.8至18.8。大腺瘤(直径超过2 cm)和腺癌患者背景正常黏膜的S期比例分别为9.74(标准差3.76%)(范围2.7 - 16.1)和8.93(标准差3.54%)(范围2.9 - 18.9)。在无任何结肠直肠肿瘤的患者正常黏膜中,S期比例为8.99(标准差3.94%)(范围3.9 - 17.7)。四组患者正常黏膜的S期比例无统计学显著差异。我们的结果表明,背景结肠隐窝增殖活性的增加并非肿瘤发生所必需。

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本文引用的文献

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Application of the crypt-isolation technique to flow-cytometric analysis of DNA content in colorectal neoplasms.
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