Variations in DNA aneuploid cell content during tumor dissociation in human colon and head and neck cancers analyzed by flow cytometry.
Variations in DNA aneuploid cell content during tumor dissociation in human colon and head and neck cancers analyzed by flow cytometry.
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通过流式细胞术分析人类结肠癌和头颈癌肿瘤解离过程中 DNA 非整倍体细胞含量的变化。
DOI：
10.1002/cyto.990140515
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发表时间：
1993
期刊：
影响因子：
--
通讯作者：
Heilbrun,LK
中科院分区：
文献类型：
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作者：
Ensley,JF;Maciorowski,Z;Hassan,M;Pietraszkiewicz,H;Sakr,W;Heilbrun,LK
涉及人实体瘤的实验研究通常需要高产量的单细胞悬浮液，其代表起源组织并且其中保留了感兴趣的细胞特性。这对于通过流式细胞术测定DNA倍性是特别必要的。机械解聚和蛋白水解酶消化是实体瘤最常用的解离技术。经常不进行技术比较测试。机械和蛋白水解酶解离技术进行了比较测试，在77人鳞状细胞癌的头部和颈部（SCCHN）和25人结肠癌的细胞产量，染料排除活力，质量和形态的DNA直方图，和DNA非整倍体亚群的存在和比例。在SCCHN的机械制备物和结肠癌的酶促制备物中注意到显著且一致的DNA非整倍体亚群损失。在机械细胞悬浮液中，SCCHN样本中DNA非整倍体亚群的频率被低估了52%，DNA非整倍体细胞的比例在另外30%的样本中减少。相反，DNA非整倍体亚群的标本的频率被低估了38%，在细胞悬液酶解离的人结肠癌和他们的比例减少了额外的50%的标本。用三种常用的蛋白水解酶孵育人结肠癌，证明DNA非整倍体亚群的逐渐丧失是酶浓度和孵育时间的函数。这是一个严重的潜在错误来源，在流式细胞术测定的DNA倍性在人类实体瘤，并可能有助于获得的结果的多样性和偶尔矛盾的结论，在这样的研究。© 1993 Wiley利斯公司
Experimental research involving human solid tumors often requires single cell suspensions of high yield that are representative of the tissue of origin and in which the cellular property of interest is preserved. This is particularly necessary for the determination of DNA ploidy by flow cytometry. Mechanical dissaggregation and proteolytic enzyme digestion are the most commonly employed dissociation techniques for solid tumors. Comparative testing of techniques is often not performed. Mechanical and proteolytic enzyme dissociation techniques were comparatively tested in 77 human squamous cell cancers of the head and neck (SCCHN) and 25 human colon cancers for cellular yield, dye exclusion viability, quality, and morphology of DNA histograms, and the presence and proportion of DNA aneuploid subpopulations. Significant and consistent DNA aneuploid subpopulation losses were noted in mechanical preparations of SCCHN and enzymatic preparations of colon cancers. The frequency of SCCHN specimens with DNA aneuploid subpopulations was underestimated by 52% in mechanical cell suspensions, and the proportion of DNA aneuploid cells was diminished in an additional 30% of the specimens. Conversely, the frequency of specimens with DNA aneuploid subpopulations was underestimated by 38% in cell suspensions from enzymatically dissociated human colon cancer and their proportion diminished in an additional 50% of the specimens. Incubations of human colon cancers with three commonly employed proteolytic enzymes demonstrated a progressive loss of DNA aneuploid subpopulations as a function of enzyme concentration and incubation time. This is a serious potential source of error in the flow cytometric determination of DNA ploidy in human solid tumors, and may contribute to the diversity of results obtained and occasional contradictory conclusions reached in such studies. © 1993 Wiley‐Liss, Inc.