摘要
目的探讨Id3在顺铂诱导人Burkitt's B淋巴瘤细胞(Daudi)增殖抑制和凋亡中的作用。方法应用顺铂处理人Daudi细胞,台盼蓝染色法检测不同浓度的顺铂对Daudi细胞生长的抑制作用;流式细胞仪(FCM)分析细胞周期和凋亡率;用RT-PCR检测顺铂处理后Id3、Id2、钙调素(CaM)基因表达水平变化。结果Daudi细胞增殖抑制率随顺铂浓度的增加而增加;2μg/ml的顺铂作用24h后,Daudi细胞表现出G1期阻滞,S期和G2/M期细胞比例下降;AnnexinⅤ/PI双染法和FCM结果表明,顺铂可诱导Daudi细胞发生凋亡;RT-PCR结果表明,顺铂处理Daudi细胞可导致Id3 mRNA的表达增加,而Id2、CaM mR-NA无明显变化。结论Id3基因的上调表达可能在顺铂抑制Daudi细胞增殖、诱导细胞凋亡中发挥作用。
Objective To explore the role of Id3 in the proliferation and apoptosis of Daudi cells stimulated with Cisplatin. Methods Daudi cells were stimulated with Cisplatin in different concentration in vitro. The inhibitory effects of cisplatin on Daudi cell proliferation were assayed with Trypan blue staining method. The cell cycle progression and apoptotic rate were determined by flow cytometry （FCM）. Expression of Id3 mRNA was detected by RT-PCR technique. Results Cell cycle analysis evaluated by PI staining method revealed that Cisplatin obviously inhibited the Daudi cell growth with dose-dependent manner. After stimulating Daudi cells with Cisplatin at the concentration of 2 μg/ml for 24 h,cell cycle of Daudi cells was apparently arrested at G1 phase. The proportion of cells in S and G2/M phases decreased. The results of Annexin V/PI staining and FCM showed that the apoptotic rate was enhanced after stimulation of Daudi cells with Cisplatin. RT-PCR indicated that the expression level of Id3 mRNA was significantly higher in Cisplatin-stimulated Daudi cells than in unstimulated cells. Conclusion Id3 gene may play an important role in the regulation of cell proliferation and apoptosis of Daudi stimulated with Cisplatin.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2007年第2期111-113,共3页
Chinese Journal of Clinical Laboratory Science
基金
江苏省"六大人才高峰"基金课题(050203D)
