摘要
目的:探讨二氟甲基鸟氨酸(dif-luoromethylornithine,DFMO)抑制多胺生物合成对人子宫内膜癌细胞系HEC-1B细胞增殖和细胞凋亡的影响。方法:HEC-1B细胞经不同浓度DFMO作用不同时间后,绘制生长曲线,观察DFMO对细胞生长的抑制作用,MTT检测细胞抑制率,用流式细胞仪(FCM)分析细胞周期变化;Giemsa染色显微照相观察凋亡细胞形态,DNALadder检测凋亡细胞DNA是否出现凋亡特征性梯形条带,FCM检测凋亡细胞百分率。结果:DFMO对人子宫内膜癌细胞HEC-1B有明显的生长抑制作用,呈剂量和时间依赖关系,2.5 mmol/L DFMO即可抑制HEC-1B细胞增殖,DFMO作用时间为5 d时,细胞增殖抑制率为60%。细胞周期分析示,DFMO使HEC-1B细胞阻滞于G1期,5 mmol/L DFMO作用2 d可使G1期细胞由68.3%增加到85.5%;5 mmol/L DFMO可诱导HEC-1B细胞凋亡,作用5 d时细胞凋亡率为63.8%,Gi-emsa染色可见凋亡小体,DNA Ladder显示典型的细胞凋亡梯形条带,加入40μmol/L外源性腐胺可阻止DFMO的作用。结论:抑制多胺生物合成可抑制子宫内膜癌细胞HEC-1B细胞增殖,诱导细胞凋亡。
OBJECTIVE: To investigate the anti proliferative and apoptosis-inductive effects of α-difluoromethylornithine (DF- MO), an irreversible inhibitor of polyamines biosynthesis, on human endometrial cancer cell line HEC-1B in vitro. METHODS: Treated with DFMO of different concentrations for differentiated time, growth characteristics of HEC 1B cells were detected through cell growth curve. The inhibition rates of cell growth were quantified by MTT assay, and the variety of cell cycle were analyzed by flow cy- tometry. The morphology of apoptotie HEC-1B cells were observed by microphotography after Wright-Giemsa staining. HEC-1B cell ap optosis were further measured by agarose gel electrophoresis and flow cytometry. RESULTS: DFMO distinctively inhibited the prolif- eration of HECqB cells in a time and dose-dependent manner. Cell growth was inhibited by 2.5 mmol/L DFMO and the inhibition rate was about 60% when HEC 1B cell were treated by DFMO for 5 d. Cell cycle analysis by flow eytometry showed that the fraction of GI phase cells increased from 68.3% to 85.5% after exposure to 5 mmol/L DFMO for 2 d, and the fact indicated that DFMO caused HEC-1B cell cycle arrest in G1 phase. Typical nuclear condensation and apoptosis bodies were observed in HEC-1B cells after 5mmol/L DFMO treatment. Apoprotic rate of DFMO-treated HEC-1B cells in- creased to 63.8% in 5 d. Genome DNA eleetrophoresis further confirmed that typical DNA ladder was formed. These cell changes were suppressed by a provision of 40 μmol/L exogenous putrescine (Pu). CONCLUSION: Polyamine depletion by DFMO can effectively inhibit cell growth and in- duce cell apoptosis in HEC-IB cells.
出处
《中华肿瘤防治杂志》
CAS
2009年第11期837-840,852,共5页
Chinese Journal of Cancer Prevention and Treatment
关键词
多胺
二氟甲基鸟氨酸
子宫内膜肿瘤
细胞增殖
细胞凋亡
poiyamines
difluoromethylornithine
endometrial neoplasms
cell proliferation
apoptosis