Pollack A, Terry N H, Wu C S, Wise B M, White R A, Meistrich M L
Department of Radiotherapy, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Cytometry. 1995 May 1;20(1):53-61. doi: 10.1002/cyto.990200109.
The simultaneous and specific staining of iododeoxyuridine (IdUrd) and chlorodeoxyuridine (CldUrd) allows for more accurate estimates of potential doubling time (Tpot). Because CldUrd is not approved for human use, the procedure was adapted for the staining of IdUrd and bromodeoxy-uridine (BrdUrd). The fluorescein isothiocyanate-conjugated B44 antibody (B44-FITC) stained both IdUrd and BrdUrd in tumor nuclei labelled singly with one or the other pyrimidine analogue. However, when MCaK tumors in exponential growth in vivo were pulse labelled with both IdUrd and BrdUrd, the staining of BrdUrd was not seen, and the labelling pattern reflected specificity to IdUrd. These observations were confirmed using tumors pulse labelled with IdUrd and/or BrdUrd at 6 h and/or 0.3 h prior to tumor removal in all possible combinations. Simultaneous specific staining of BrdUrd by Br3 and of IdUrd by B44-FITC was documented by quantification of labelling indices (LIs) from double-labelled tumors. The specificity of B44-FITC for IdUrd in double-labelled tumors was due to a greater affinity of this antibody for IdUrd than for BrdUrd. This technique allowed for two independent estimates of LI and Tpot when tumors were double labelled for 3.0 and 5.5 h. Both IdUrd and BrdUrd are approved for clinical use, and this double-labelling technique should prove to be valuable for measuring the cell kinetics of solid tumors in vivo.
对碘脱氧尿苷（IdUrd）和氯脱氧尿苷（CldUrd）进行同步特异性染色，能够更准确地估计潜在倍增时间（Tpot）。由于CldUrd未获批准用于人体，因此对该程序进行了调整，以用于IdUrd和溴脱氧尿苷（BrdUrd）的染色。异硫氰酸荧光素偶联的B44抗体（B44-FITC）可对用一种嘧啶类似物单独标记的肿瘤细胞核中的IdUrd和BrdUrd进行染色。然而，当体内呈指数生长的MCaK肿瘤用IdUrd和BrdUrd进行脉冲标记时，未观察到BrdUrd的染色，且标记模式显示对IdUrd具有特异性。在肿瘤切除前6小时和/或0.3小时，以所有可能的组合用IdUrd和/或BrdUrd对肿瘤进行脉冲标记，这些观察结果得到了证实。通过对双标记肿瘤的标记指数（LI）进行定量，记录了BrdUrd被Br3同时特异性染色以及IdUrd被B44-FITC特异性染色的情况。在双标记肿瘤中，B44-FITC对IdUrd的特异性是由于该抗体对IdUrd的亲和力高于对BrdUrd的亲和力。当肿瘤进行3.0小时和5.5小时的双标记时，该技术可对LI和Tpot进行两个独立的估计。IdUrd和BrdUrd均已获批用于临床，这种双标记技术对于测量体内实体瘤的细胞动力学应具有重要价值。
