摘要
目的观察南赤瓟提取物对宫颈癌HeLa细胞凋亡的诱导作用,并探讨其可能的作用机制。方法用不同浓度的南赤瓟提取物(0.10、0.25、0.50、1.00、2.00 mg/ml)于不同作用时间(24、48、72 h)处理宫颈癌HeLa细胞后,采用MTT法检测不同浓度的南赤瓟提取物在不同作用时间下对HeLa细胞增殖率的影响;用0.50和1.00 mg/ml的南赤瓟提取物处理宫颈癌HeLa细胞48 h后Hoechst33258染色观察细胞形态变化,Annexin V-FITC/PI染色测定细胞凋亡率,JC-1染色法测定细胞线粒体跨膜电位水平,Caspase试剂盒检测HeLa细胞中Caspase-9和Caspase-3的活性,荧光定量PCR测定Bcl-2和Bax mRNA表达,并计算Bcl-2与Bax的比值。结果南赤瓟提取物对HeLa细胞的生长抑制作用呈明显的时间-剂量依赖性;南赤瓟提取物(0.50和1.00 mg/ml)处理能显著降低HeLa细胞线粒体膜电位,促进HeLa细胞凋亡(P<0.05,P<0.01);上调Bax mRNA表达,下调Bcl-2 mRNA表达和Bcl-2与Bax的比值以及增强Caspase-9和Caspase-3的活性(P<0.05,P<0.01)。结论南赤瓟提取物可诱导HeLa细胞凋亡,其作用机制与降低HeLa细胞线粒体膜电位及调节凋亡相关基因Bcl-2、Bax和Caspase-9、Caspase-3的表达有关。
Objective To investigate the effect of Thldiantha nudiflora Hemsl extract on the apoptosis in human cervix cancer HeLa cells. Methods HeLa cells were treated with Thldiantha nudiflora Hemsl extract at different concentrations of 0.10, 0.25, 0.50, 1.00, and 2.00 mg/ml for different times （24, 48, and 72 h）. Cell viability was determined by MTT assay. After HeLa cells were treated with Thldiantha nudiflora Hemsl extract （0.5 and 1.0 mg/ml）, cell morphology was examined by hoechst33258 through fluorescence microsco- py. Annexin V/PI double-staining was performed to detect apoptosis of HeLa cells by flow cytometry. The mito- chondrial membrane potential of HeLa cells was measured by JC-1 staining. The activities of Caspase-9 and Caspase-3 were measured by Caspase assay kit. The expression of Bcl-2 and Bax at mRNA level was detected by real-time PCR, and Bcl-2/Bax ratio was analyzed. Results The inhibitory effect of Thldiantha nudiflora Hemsl extract on HeLa cells showed a significant time-dose response. Thldiantha nudiflora Hemsl extract （0.50 and 1.00 mg/ml） significantly decreased the mitochondrial membrane potential of HeLa cells, promoteed apop- tosis（P 〈0.05, P 〈 0. 01, respectively）, up-regulated the mRNA expression of Bax, down-regulated the ex- pression of Bcl-2 and Bcl-2/Bax ratio, and increased the activities of Caspase-9 and Caspase-3, when compared with the negative control group （P 〈 0. 05, P 〈 0. 01 ）. Conclusion Thldiantha nudiflora Hemsl extract effec- tively induces the apoptosis in HeLa cells. The mechanism may be related to decreasing the mitochondrial mem- brane potential of HeLa ceils, and regulating the expression of apoptosis-related genes Bcl-2, Bax, Caspase-9, and Caspase-3.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2012年第18期1844-1848,共5页
Journal of Third Military Medical University
基金
湖北省卫生厅中医药中西医结合科研项目(2010Z-Y09)~~
关键词
南赤瓟
宫颈癌
HELA细胞
凋亡
Thldiantha nudiflora Hemsl
cervix cancer
HeLa cells
apoptosis
