title: Application of the crypt isolation technique to the flow cytometric analysis of DNA content in gastric carcinoma.
abstract: An inevitable limitation of conventional flow cytometric analysis of gastric cancer DNA content is that the preparations of tumor cell nuclei are contaminated with stromal cell nuclei. Using the crypt isolation technique, we separated tumor tissues from stromal tissues and analyzed the DNA content in samples of pure gastric cancer cells (64 intestinal-type and 46 diffuse-type) by flow cytometry. Morphologically, crypts from well-differentiated and moderately differentiated adenocarcinomas usually showed large tube-like or sheet structures, whereas tumor tissues isolated from poorly differentiated adenocarcinomas usually exhibited small tumor cell clumps or clusters of varying sizes. Tumor ploidy was divided into diploid, aneuploid, and multiploid subgroups. Aneuploidy and multiploidy were observed in 12% (13 of 110) and 64% (71 of 110) of gastric cancers, respectively. A high frequency of DNA aneuploidy or multiploidy was associated with intestinal-type tumors, but not with any of the other clinicopathologic variables tested. In contrast, high S-phase fraction values demonstrated a close association with tumors with abnormal ploidy, advanced stage, intestinal type, and late TNM stage. Our results suggest that S-phase fraction may be a more useful indicator of aggressive behavior in gastric cancers than DNA aneuploidy. To our knowledge, the present study is the first to report flow cytometric DNA content in a large number of gastric cancer samples obtained using the crypt isolation technique.
authors.affiliation: Division of Pathology, Central Clinical Laboratory, School of Medicine, Iwate Medical University, Uchimaru, Morioka, Japan.
authors.forename: Yu-Fei
authors.initials: YF
authors.lastname: Jiao
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Sugai
authors.forename: Masamichi
authors.initials: M
authors.lastname: Suzuki
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Uesugi
authors.forename: Wataru
authors.initials: W
authors.lastname: Habano
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Nakamura
authors.forename: Akinori
authors.initials: A
authors.lastname: Takagane
authors.forename: Takayuki
authors.initials: T
authors.lastname: Suto
authors.forename: Toru
authors.initials: T
authors.lastname: Yoshida
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000473:pathology; D000328:Adult; D000368:Aged; D004273:DNA, Neoplasm / Q000032:analysis*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D008297:Male; D008875:Middle Aged; D009367:Neoplasm Staging; D011003:Ploidies; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology
chemical_list: D004273:DNA, Neoplasm