呋喃唑酮对Hep G2肝癌细胞增殖和细胞周期的影响论著

The Effects of Furazolidoze on Cell Proliferation and Cell Cycle of Hep G2

背景与目的：探讨呋喃唑酮（furazolidoze，FZD）对体外培养的人肝癌细胞（HepG2）的毒性机制。材料与方法：分别以不同浓度FZD（0．00、0．78、1．56、3．12、6．25、12．50、25．00、50．00μg／m1）作用HepG2细胞24h，采用噻唑蓝（MTT）比色法测定FZD对细胞增殖的影响；流式细胞术（FCM）检测细胞周期的分布；RT-PCR半定量检测S期关卡相关基因的变化。结果：随着FZD浓度的增加，HepG2细胞的存活率逐渐降低，当浓度超过6．25μg／ml时，细胞存活率下降显著（P〈0．05）；细胞生长曲线显示各染毒浓度组细胞生长速度明显减慢；细胞周期动力学显示FZD在0～3．12μg／ml范围内s期细胞比例明显增加，G-期细胞比例下降；RT-PCR结果显示FZD作用后p2J呈高表达，cyclinE,cyclinA和Cdk2表达量则不同程度下调。结论：FZD对体外培养的HepG2细胞有一定的细胞毒性且诱导s期阻滞，其增殖抑制作用可能是通过激活s期损伤关卡实现的。

BACKGROUND AND AIM： To investigate the mechanisms for cytotoxicity of furazolidoze （FZD） to human hepatocarcinoma cell line （Hep G2） .MATERIALS AND METHODS： After Hep G2 cells were exposed to FZD at the different concentrations（0, 0.78, 1.56, 3.12, 6.25, 12.50, 25.00,50.00 μg/ml）for 24 h, the proliferation effect was measured by MTT assay whilst the cell cycle distribution was assessed by flow cytometry. The mRNA of S stage checkpoint was evaluated by RT-PCR. RESULTS： MTT assay showed that Hep G2 survival rate decreased with increasing FZD concentration, whereas the survival rate was significantly higher with concentration above 6.25 μg/ml（P 〈 0.05）. Cell growth assay showed that the growth of Hep G2 was significantly slower than control. Treated with FZD （0-3.12 μg/ml）, cells in S stage increased, while those in G1 stage decreased. The expression level of p21 increased and the expression of cyclinE, cyclinA , Cdk2 decreased after treatment with FZD. CONCLUSION： FZD exhibited cytotoxicity to Hep G2 in vitro and induced cell arrest at S stage, and it may exert antiproliferative effect through activation of the S stage damage checkpoint.