摘要
目的观察人参皂甙单体Rh2(GS-Rh2)对人脑胶质瘤U251细胞增殖和凋亡的影响,并探讨其机制。方法用GS-Rh2处理人脑胶质瘤U251细胞,采用MTT法检测U251细胞增殖抑制率,流式细胞仪和Annexin V-FITC/PI双染法观察U251细胞凋亡情况,TRAP-ELISA法检测U251细胞端粒酶活性,RT-PCR法检测U251细胞人端粒酶逆转录酶(hTERT)mRNA的表达。结果 5、10、20、40、80μg/ml GS-Rh2组U251细胞增殖抑制率分别为20.42%、32.19%、45.09%、57.37%、73.82%。根据细胞增殖抑制曲线计算出IC30、IC50、IC70值分别为9.7、25.2、68.4μg/ml。FITC和PI荧光双参数点图显示实验组细胞分布区域与对照组相比出现明显改变,随着药物浓度的增加,早期凋亡、晚期凋亡或坏死的区域细胞数量逐渐增多。9.7、25.2、68.4μg/ml GS-Rh2处理12、24、48、72 h后U251细胞端粒酶活性随GS-Rh2浓度增加和作用时间的延长而降低。浓度为25.2μg/ml GS-Rh2作用24、48、72 h的U251细胞hTERTmRNA与β-actin灰度值比为0.964 9±0.004 2、0.401 8±0.001 4、0.297 8±0.001 8,对照组为0.976 4±0.005 1。作用48、72 h U251细胞hTERT mRNA与β-actin灰度值比与对照组相比P均<0.05;作用48、72 h者相比P<0.05。结论 GS-Rh2能够诱导人脑胶质瘤U25l细胞凋亡,抑制U25l细胞增殖。其机制与其抑制hTERT基因表达,降低端粒酶活性有关。
Objective To observe the effect and mechanism of ginsenoside-Rh2 （GS-Rh2 ） on human glioma U251 cell proliferation and apoptosis. Methods Human glioma U251 cells were treated with GS-Rh2 ,the inhibitory effect was examined with MTT. Staining with Annexin V-FITC/PI and flow cytometry analysis were used to detect the apoptotic cells. TRAP-ELISA was used to detected telomerase activity and RT-PCR was used to detect the expression of hTERT gene. Resuits The apoptotic rate was 20.42% in the group with 5 μg/ml GS-Rh2 ,32.19% in the group with 10 μg/ml GS-Rh2, 45.09% in the group with 20 μg/ml GS-Rh2,57.37% in the group with 40 μg/ml GS-Rh2, 73.82% in the group with 80 μg/ml GS-Rh2. The value of IC30, IC50, IC70 were 9.7,25.2,68.4 μg/ml respectively. FITC and PI fluorescence dual-parameter point map showed that there was a significantly change between the regional distribution of experimental cells compared with control group, and early apoptosis, late apoptosis or necrosis of regional cells gradually increased with the increase of drug concentration. The telomerase activity of U251 cells which treated with 9.7,25.2,68.4μg/ml GS-Rh2 decreased with the concentration of GS-Rh2 and the treatment time. The grey ratio of hTERT mRNA and β-actin in U251 cells treated with 25.2μg/ml GS-Rh2 for 24,48,72 h were 0. 964 9 ±0. 004 2,0.401 8 ±0.1201 4,0. 297 8±0. 001 8 respectively,and it was 0. 976 4 ± 0.005 1 in control group. Conclusions GS-Rh2 can induce human glioma U251 cell apoptosis,inhibite cell proliferation. It may be related to GS-Rh2 decreasing hTERT gene expression and inhibiting telomerase activity.
出处
《山东医药》
CAS
北大核心
2010年第22期16-18,共3页
Shandong Medical Journal
基金
辽宁省教育厅科研基金资助项目(2009A448)
