Saito S, Kagabu T, Nishijima M, Nishiya I
Department of Obstetrics and Gynecology, Iwate Medical University, Morioka.
Nihon Sanka Fujinka Gakkai Zasshi. 1990 Mar;42(3):227-33.
We analyzed the characterization of DNA synthesized cells (S phase cells) in the proliferation. It is important that the fixation and cell cycle time of S phase cells and DNA qualitative changes, etc., be examined. We used the dual-laser FCM system of the Los Alamos National Laboratory in New Mexico and analyzed the cell cycle, especially changes in S phase cells both quantitatively and qualitatively. By the bromodeoxyuridine (BrdU)/Hoechst Quenching effect, we could detect differences in electric signals by adopting the differential fluorescence correction method. (1) The effects of sodium butyrate in gene expression were dose-dependent at human endometrial adenocarcinoma cells. (2) After double staining with Mithramycin and Hoechst33342, S phase cells incorporating BrdU under quenching gave from linear relative to arched bivariate contour histograms. (3) At 24 hours G1 phase synchronization of SB 1,3 and 5mM appeared in slow cycling cells after BrdU was added. These results suggest that the effect of sodium butyrate in human endometrial adenocarcinoma cells might be understood not only from the quantitative change, but also by the qualitative change in S phase cells.
我们分析了增殖过程中DNA合成细胞(S期细胞)的特征。检查S期细胞的固定、细胞周期时间以及DNA定性变化等非常重要。我们使用了新墨西哥州洛斯阿拉莫斯国家实验室的双激光流式细胞术系统,对细胞周期,尤其是S期细胞的变化进行了定量和定性分析。通过溴脱氧尿苷(BrdU)/Hoechst淬灭效应,我们采用差分荧光校正法检测电信号的差异。(1)丁酸钠对人子宫内膜腺癌细胞基因表达的影响呈剂量依赖性。(2)用光辉霉素和Hoechst33342双重染色后,在淬灭条件下掺入BrdU的S期细胞产生的双变量轮廓直方图从线性变为拱形。(3)添加BrdU后,在慢循环细胞中,24小时时1、3和5mM的丁酸钠使G1期同步化。这些结果表明,丁酸钠对人子宫内膜腺癌细胞的作用不仅可以从数量变化来理解,还可以通过S期细胞的质量变化来理解。