Detection of cells resistant to alkylating agents by flow cytometric analysis of DNA damage.
Detection of cells resistant to alkylating agents by flow cytometric analysis of DNA damage.
复制标题
通过 DNA 损伤的流式细胞分析检测细胞对烷化剂的耐药性。
DOI：
10.1002/cyto.990110807
复制
发表时间：
1990
期刊：
影响因子：
--
通讯作者：
Sugarbaker,EV
中科院分区：
文献类型：
--
作者：
Frankfurt,OS;Seckinger,D;Sugarbaker,EV
用流式细胞术分析对烷基化剂敏感和耐药的细胞的DNA损伤情况。用该药物处理人卵巢癌细胞系A2780和对L‐苯丙氨酸芥菜（L‐PAM）抗性高7倍的亚系，固定，并用单克隆抗体（MOAB） F7‐26染色，该抗体检测烷基化DNA的单链区域。流式细胞仪测定平均荧光强度。染色前加热细胞，扩增烷基化DNA的单链性。与耐药细胞相比，敏感细胞与DNA的结合量显著增加。每νg L‐PAM不敏感细胞荧光增加80个通道，抗性细胞荧光仅增加17个通道。敏感和耐药细胞分别用L - PAM处理，按不同比例混合，并用MOAB染色。在荧光直方图上，敏感细胞和耐药细胞的荧光强度有十多年的差异。在敏感细胞中检测到2-5%的耐药细胞作为一个单独的细胞亚群。我们得出结论，用MOAB F7‐26染色可以作为细胞对烷基化剂敏感性或抗性的指标。通过流式细胞仪分析在异质群体中检测少量耐药细胞亚群可能有助于监测新出现的耐药性。
DNA damage was measured by flow cytometric analysis of cells sensitive and resistant to alkylating agents. Human ovarian carcinoma cell line A2780 and a subline which is 7 times more resistant to L‐phenylalanine mustard (L‐PAM) were treated with the drug, fixed, and stained with monoclonal antibody (MOAB) F7‐26 which detects single‐stranded regions in alkylated DNA. Mean fluorescent intensity was measured on a flow cytometer. Cells were heated before staining to amplify single‐strandedness in alkylated DNA. Significantly larger amount of MOAB was bound to DNA in sensitive than in resistant cells. Fluorescence increased by 80 channels per νg L‐PAM insensitive cells and only by 17 channels in resistant cells. Sensitive and resistant cells were treated with L‐PAM, mixed in different proportions, and stained with MOAB. Populations of sensitive and resistant cells were clearly separated on fluorescence histograms by more than a decade difference in fluorescence intensity. Presence of 2–5% resistant cells was detected among sensitive cells as a separate cell subset.We conclude that staining with MOAB F7‐26 can be used as an indicator of cell sensitivity or resistance to alkylating agents. Detection of minor subsets of resistant cells in heterogeneous populations by FCM analysis may be useful for monitoring emerging drug resistance.
登录
查看更多内容
顺式和反式二氯二氨铂 (II) 与脱氧核糖核酸的结合会暴露核苷（用抗核苷抗体进行免疫化学测量）。
影响因子：
2.9
作者：
Sundquist,WI;Lippard,SJ;Stollar,BD
通讯作者：
Stollar,BD
影响因子：
7.8
作者：
Darzynkiewicz, Z;Traganos, F;Sharpless, T;Melamed, M R
通讯作者：
Melamed, M R
链脲佐菌素烷基化对 DNA 二级结构的影响
影响因子：
4
作者：
R. Rizvi;Shahabuddin;A. Rahman;S. M. Hadi
通讯作者：
S. M. Hadi
通过吖啶橙染色和自动细胞荧光测定法研究 DNA 原位热变性。
影响因子：
3.7
作者：
Zbigniew Darzynkiewicz;F. Traganos;T. Sharpless;Myron R. Melamed
通讯作者：
Myron R. Melamed
DNA损伤的流式细胞术分析和烷化剂的细胞毒性评价。
影响因子：
11.2
作者：
Frankfurt,OS
通讯作者：
Frankfurt,OS