Peters G J, Laurensse E, Leyva A, Pinedo H M
FEBS Lett. 1984 May 21;170(2):277-80. doi: 10.1016/0014-5793(84)81328-9.
The concentration of PRPP (phosphoribosyl pyrophosphate) measured in tumor cells grown in monolayer showed a large variation with the various harvesting methods examined, including trypsinization. This variation could be reduced by a 1-h incubation of trypsinized cells as a suspension in Dulbecco's medium. After this preincubation these cell suspensions were suitable for the study of modulation of PRPP. One microM methotrexate caused a 2-3-fold increase and 1 mM N- phosphonoacetyl -L-aspartate a slight increase, but inosine and deoxyinosine drastically reduced PRPP concentrations. 5-Fluorouracil had no effect. This study demonstrates that metabolic parameters such as PRPP concentrations can be studied conveniently in suspensions of cells which are commonly cultivated as monolayers.
在单层培养的肿瘤细胞中测得的5-磷酸核糖焦磷酸(PRPP)浓度,在包括胰蛋白酶消化法在内的各种检测收获方法下显示出很大差异。将胰蛋白酶消化后的细胞以悬浮液形式在杜尔贝科培养基中孵育1小时,这种差异可减小。预孵育后,这些细胞悬浮液适用于PRPP调节的研究。1微摩尔甲氨蝶呤使PRPP浓度增加2至3倍,1毫摩尔N-膦酰乙酰-L-天冬氨酸使PRPP浓度略有增加,但肌苷和脱氧肌苷可大幅降低PRPP浓度。5-氟尿嘧啶无作用。本研究表明,像PRPP浓度这样的代谢参数可以在通常以单层形式培养的细胞悬浮液中方便地进行研究。