摘要
本研究旨在探讨齐墩果酸诱导人T淋巴细胞白血病Jurkat细胞株凋亡及对PTEN表达的影响。应用CCK-8法检测细胞增殖抑制率,用Hoechst33258染色观察凋亡细胞形态,Annexin V/PI双染色后流式细胞仪检测细胞凋亡,并应用实时定量RT-PCR和Western blot方法分别检测PTEN基因及其蛋白表达水平。结果表明:齐墩果酸以时间和剂量依赖方式抑制Jurkat细胞增殖,12、24和48小时的半数抑制浓度(IC50)分别约为85.35、53.66和33.18μmol/L。流式细胞术检测显示,齐墩果酸以0、40、80和160μmol/L浓度作用细胞24小时凋亡率分别为6.72%、19.8%、28.72%和30.12%(p<0.05)。80和160μmol/L齐墩果酸分别处理Jurkat细胞24小时后PTEN-mRNA及蛋白表达上调。结论:PTEN基因和蛋白表达上调可能参与齐墩果酸对Jurkat细胞的抑制增殖和诱导凋亡作用。
This study was aimed to explore the effects of oleanlic acid on PTEN expression and apoptosis of Jurkat cells.The inhibitory rate was measured by Cell Counting Kit-8.The apoptotic nucleus morphous was observed by Hoechst 33258 staining.The apoptosis rate of Jurkat cells were determined by flow cytometry with Annexin V/PI double staining.PTEN mRNA and protein were detected by quantitative real-time PCR and Western blot respectively.The results showed that oleanlic acid inhibited the proliferation of Jurkat cells in time-and dose-dependent manners.The 50% growth inhibition（IC50） at 12,24 and 48 hours were about 85.35 μmol/L,53.66 μmol/L and 33.18 μmol/L respectively.Flow cytometric assay showed that the apoptotic rates of Jurkat cells treated with oleanlic acid（0,40,80 and 160 μmol/L） for 24 hours were 6.72%,19.8%,28.72% and 30.12%（p〈0.05）.PTEN mRNA and protein expressions were up-regulated in Jurkat cells treated with oleanlic acid of concentration 80 μmol/L and 160 μmol/L for 24 hours.It is concluded that up-regulation of PTEN mRNA and PTEN protein may be involved in oleanlic acid-induced Jurkat cell apoptosis.
出处
《中国实验血液学杂志》
CAS
CSCD
2011年第2期367-371,共5页
Journal of Experimental Hematology
基金
沈阳市高技术产业发展项目(编号沈发改发2010-106号)
辽宁省科学技术计划(编号2010225032)
