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

Abstract:

BACKGROUND/AIMS:Conventional flow-cytometric analysis of colorectal tumor DNA content includes both tumor and stromal cell DNA. To improve the method, we evaluated the DNA content of purified epithelial cells using the crypt-isolation technique. METHODS:Normal and neoplastic crypts were isolated from fresh resected colorectum nonenzymatically. Isolated crypts were digested into single nuclei suspensions and examined by flow cytometry (FCM). Internal controls from normal crypts were used to define diploidy. The neoplastic nuclei in the same individual and mixtures of both normal and neoplastic nuclei in various ratios were analyzed. RESULTS:Tumors having both diploid and aneuploid stemlines were found more frequently than that reported using conventional FCM. Near-diploid DNA stemlines with DNA indices ranging between 0.90 and 1.10 were detectable. The cases of advanced cancer in which the DNA index was between 0.90 and 0.95 were associated significantly with Dukes' stage C. CONCLUSIONS:This is the first reported analysis of the DNA content of "pure" neoplastic colorectal epithelia using FCM. Multiploid and near-diploid stemlines were more accurately analyzed than that using conventional FCM.

journal_name

Gastroenterology

journal_title

Gastroenterology

authors

Nakamura S,Goto J,Kitayama M,Kino I

doi

10.1016/s0016-5085(94)94651-5

subject

Has Abstract

pub_date

1994-01-01 00:00:00

pages

100-7

issue

1

eissn

0016-5085

issn

1528-0012

pii

S0016-5085(94)94651-5

journal_volume

106

pub_type

杂志文章
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    pub_type: 杂志文章,评审

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    doi:

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    doi:

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    doi:

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    doi:

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    doi:

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