摘要
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.
出处
《癌变.畸变.突变》
CAS
CSCD
2008年第3期201-204,共4页
Carcinogenesis,Teratogenesis & Mutagenesis