Gray J W, Pallavicini M G, George Y S, Groppi V, Look M, Dean P N
J Cell Physiol. 1981 Aug;108(2):135-44. doi: 10.1002/jcp.1041080204.
We report measurements of the incorporation of radioactive molecules during short labeling periods, as a function of cell-cycle stage, using a cell-sorter-based technique that does not require cell synchronization. We have determined: (1) tritiated thymidine (3H-TdR) incorporation throughout S-phase in Lewis lung tumor cells in vitro both before and after treatment with cytosine arabinoside; (2) 3H-TdR incorporation throughout S-phase in KHT tumor cells in vitro and in vivo; (3) 3H-TdR incorporation throughout S-phase in Chinese hamster ovary cells and compared it with DNA synthesis throughout S-phase; (4) a mathematical expression describing 3H-TdR incorporation throughout S-phase in Chinese hamster M3-1 cells; and (5) the simultaneous incorporation of 3H-TdR and 35S-methionine as they are related to cell size and DNA content in S49 mouse lymphoma cells. In asynchronously growing cells in vitro and in vivo, 3H-TdR incorporation was generally low in early and late S-phase and highest in mid-S-phase. However, in Lewis lung tumor cells treated with cytosine arabinoside 3H-TdR incorporation was highest in early and late S-phase and lowest in mid-S-phase. Incorporation of 35S-methionine increased continuously with cell size and DNA content. Incorporation of 3H-TdR in CHO cells was proportional to DNA synthesis.
我们报告了使用基于细胞分选仪的技术在短标记期内对放射性分子掺入情况的测量,该技术不需要细胞同步化,测量结果是细胞周期阶段的函数。我们已经确定:(1)在体外培养的Lewis肺癌细胞中,在用阿糖胞苷处理前后,整个S期的氚标记胸腺嘧啶核苷(3H-TdR)掺入情况;(2)体外和体内KHT肿瘤细胞整个S期的3H-TdR掺入情况;(3)中国仓鼠卵巢细胞整个S期的3H-TdR掺入情况,并将其与整个S期的DNA合成情况进行比较;(4)描述中国仓鼠M3-1细胞整个S期3H-TdR掺入情况的数学表达式;(5)S49小鼠淋巴瘤细胞中3H-TdR和35S-甲硫氨酸的同时掺入情况及其与细胞大小和DNA含量的关系。在体外和体内异步生长的细胞中,3H-TdR掺入在S期早期和晚期通常较低,在S期中期最高。然而,在用阿糖胞苷处理的Lewis肺癌细胞中,3H-TdR掺入在S期早期和晚期最高,在S期中期最低。35S-甲硫氨酸的掺入随着细胞大小和DNA含量的增加而持续增加。CHO细胞中3H-TdR的掺入与DNA合成成正比。