As_2O_3对淋巴瘤Raji细胞抑制效应及其Id4基因表达的影响

Depressive effect and expression of Id4 mRNA of arsenic trioxide on Raji lymphoma cells

目的:探讨As2O3对恶性淋巴瘤细胞的作用和对DNA结合抑制因子(Id4)基因表达的影响。方法:体外培养人Burkitt’s淋巴瘤细胞株Raji细胞,设对照组和用不同浓度的As2O3作用不同时间处理Raji细胞的实验组。MTT比色法检测细胞生长;流式细胞术(FCM)检测细胞凋亡并分析As2O3对细胞周期的影响。逆转录聚合酶链反应(RT-PCR)法检测处理前后Id4 mRNA的表达情况。结果:实验组MTT比色实验结果表明,As2O3各浓度组作用于Raji细胞24、48及72h后抑制率为12.15%～92.17%,P<0.05;流式细胞术检测细胞凋亡结果显示,实验组As2O3各浓度组作用于Raji细胞48h后出现明显的凋亡峰,对照组未见凋亡峰或仅有低平的凋亡峰;细胞周期分析结果显示,实验组As2O3各浓度组作用于Raji细胞48h后,As2O3可阻滞Raji细胞于G0/G1期,且随着As2O3浓度的增加,阻滞作用增强,呈一定剂量依赖关系。随药物浓度增加,Raji细胞中Id4基因mRNA的相对表达量逐渐增加,呈一定剂量依赖关系。结论:As2O3体外对Raji淋巴瘤细胞生长具有明显的抑制作用;As2O3能诱导Raji细胞凋亡,且使G0/G1期细胞比例增加,同时S及G2/M期细胞比例减少。随着As2O3浓度增加Id4基因出现表达。

OBJECTIVE：To study the effect of arsenic trioxide on the treatment of malignancy lymphoma and Id4. METHODS：Human Burkitt＇s Raji lymphoma cells were cultivated in vitro. Two groups were set up as the control group and the experiment group that was treated with various concentrations and time of arsenic trioxide. The growth of the cells was tested by MTT colorimetry,and the apoptosis and cycle distribution of cells affected by arsenic trioxide were detected by flow cytometry （FCM）. The expression of Id4 mRNA in Raji cells treated with arsenic trioxide was detected by RT-PCR. RESULTS：MTT showed that in the experimental group,the cell inhibitive rates of Raji after treated with various concentrations of arsenic trioxide for 24,48,72 hours were 12.15%-92.17%（P0.05）. FCM showed that apoptosis peak emerged significantly after Raji cells treated with arsenic trioxide for 48 hours in the experimental group,but there was no or only a low apoptosis peak in the control group. FCM showed that after Raji cells treated with various concentrations of arsenic trioxide for 48 hours in the experiment group,the cell percentage in G0/G1 phase,S phase,G2/M phase of the cell cycle distribution existed a significant difference compared with the control group. Arsenic trioxide could arrest cell cycle,the stoppage increased gradually with arsenic trioxide concentrations increasing,which was dose-deendent. As compared with the β-actin,the expression of Id4 mRNA in Raji cells after treated with arsenic trioxide increased gradually with arsenic trioxide concentrations increasing,which was also dose-dependent. CONCLUSIONS：The Raji cells are depressed effectively by arsenic trioxide in vitro. Arsenic trioxide can induce the apoptosis in Raji cells,with the increase of Raji cells in G0/G1 phase,and decrease of Raji cells in S and G2/M phase. Arsenic trioxide can recover the expression of Id4 mRNA with a dose dependent manner.