A well-known suicide gene therapy approach, cytosine deaminase (CD) in combination with prodrug 5-flurocytosine (5-FC), has become an effective strategy of tumor treatment. However, there are short of simple and convenient detection methods to evaluate the efficiency of 5-FC conversion to 5-fluorouracil (5-FU) in human cells carrying various CD/5-FC systems. In this study, we developed an effective capillary zone electrophoresis (CZE) method to simultaneously measure 5-FC and 5-FU in cells carrying CD/5-FC suicide gene system. Under the condition of 60 mM borate buffer (pH 9.5) and 25 kV separation voltage with 0.5 psi × 15 s injection in 210 nm, the separation of 5-FC and 5-FU could be completely achieved within 15 min. The linearity of the calibration curve of standard 5-FC and 5-FU was in the range from 1 to 1000 μM (r² > 0.999) and their recoveries were 98.4% and 96.0%, respectively. Due to the simple sample preparation and easy detection, this method is suitable for the study of the conversion efficiency of CD/5-FC suicide gene system. It aims to intuitively evaluate CD/5-FC systems and helps to guide the improvement of more effective CD/5-FC suicide gene systems.

胞嘧啶脱氨酶（CD）与前药5-氟胞嘧啶（5-FC）结合是一种著名的自杀基因治疗方法，已成为一种有效的肿瘤治疗策略。然而，在携带各种CD / 5-FC系统的人类细胞中，缺乏简单，方便的检测方法来评估5-FC转化为5-氟尿嘧啶（5-FU）的效率。在这项研究中，我们开发了一种有效的毛细管区带电泳（CZE）方法，可同时测量携带CD / 5-FC自杀基因系统的细胞中的5-FC和5-FU。在60 mM硼酸盐缓冲液（pH 9.5）和25 kV分离电压（在210 nm内注入0.5 psi×15 s）的条件下，可以在15分钟内完全分离出5-FC和5-FU。标准5-FC和5-FU的校准曲线的线性范围为1至1000μM（r ² > 0.999），其回收率分别为98.4％和96.0％。由于样品制备简单，易于检测，该方法适用于研究CD / 5-FC自杀基因系统的转化效率。它旨在直观地评估CD / 5-FC系统，并帮助指导改进更有效的CD / 5-FC自杀基因系统。