Chen Hong-ying, Zheng Cong-yi, Zou Guo-lin, Xie Da-xing, Gong Jian-ping
College of Life Sciences, Wuhan University, Wuhan 430072, China.
Acta Pharmacol Sin. 2004 Dec;25(12):1698-704.
To investigate the mechanism of peplomycin (PEP)-induced apoptosis in liver carcinoma cell line (Bel-7402).
Growth inhibition by PEP was analyzed using 3- 4,5-Dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptotic cells were detected using Hoechest 33258 staining, and confirmed by flow cytometric analysis and DNA fragmentation analysis. The expression of cyclin A and B1 were determined by flow cytometry and Western blot. Annexin V assay was measured by flow cytometric analysis.
PEP induced apoptosis and then inhibited cell proliferation in liver carcinoma cell line Bel-7402. Cells treated with PEP 50 mumol/L for 15 h were arrested in G2-phase with dramatical expression of cyclin A and a little change in cyclin B1. Almost all the apoptosis occurred in cells undergoing the G1-phase after treatment for 24 h.
Peplomycin induced G1-phase specific apoptosis in Bel-7402 involving G2-phase arrest.
研究培普利霉素（PEP）诱导肝癌细胞系（Bel-7402）凋亡的机制。
采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法分析PEP对细胞生长的抑制作用。使用Hoechest 33258染色检测凋亡细胞，并通过流式细胞术分析和DNA片段化分析进行确认。通过流式细胞术和蛋白质免疫印迹法测定细胞周期蛋白A和B1的表达。采用流式细胞术分析膜联蛋白V检测法。
PEP诱导肝癌细胞系Bel-7402凋亡，进而抑制细胞增殖。用50 μmol/L PEP处理15小时的细胞停滞于G2期，细胞周期蛋白A表达显著增加，细胞周期蛋白B1变化不大。处理24小时后，几乎所有凋亡都发生在处于G1期的细胞中。
培普利霉素诱导Bel-7402细胞发生G1期特异性凋亡，涉及G2期停滞。
