薤白总皂苷对人宫颈癌HeLa细胞增殖与凋亡作用的影响

目的观察薤白总皂苷对宫颈癌HeLa细胞增殖与凋亡作用的影响,并探讨其可能的作用机制。方法用不同浓度的薤白总皂苷(25、50、100、250、500、1000μg/ml)于不同作用时间(24、48、72 h)处理宫颈癌HeLa细胞后,采用MTT法检测不同浓度的薤白总皂苷在不同作用时间下对HeLa细胞增殖率的影响;用100和200μg/ml的薤白总皂苷处理宫颈癌HeLa细胞48 h后,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。结果薤白总皂苷对HeLa细胞的生长抑制作用呈明显的时间一剂量依赖性;薤白总皂苷(100和200μ/ml)处理能显著降低HeLa细胞线粒体膜电位,抑制HeILa细胞增殖,促进其凋亡(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)。结论薤白总皂苷有显著的抑制HeLa细胞增殖和诱导凋亡作用,其作用机制可能与其降低HeLa细胞线粒体膜电位,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的活性有关。

Effect of total saponin from allium macrostemon bunge on proliferation and apoptosis of cervix cancer HeLa cells

Objective To investigate the effect of total saponin from allium macrostemon bunge on proliferation and apoptosis of cervix cancer HeLa cells,and to explore the possible mechanism.Methods The HeLa cells were dealed with different concentration of total saponin from allium macrostemon bunge(25,50,100,250,500,1000μg/ml) at different time points(24,48,72h).The cell proliferation was determined by MTT assay.After HeLa cells were treated with total saponin from Allium macrostemon Bunge(100 and 200μg/ml),and the Annexin V-FITC/PI double staining was performed to detect apoptosis of HeLa cells with flow cytometry;The mitochondrial membrane potential of HeLa cells was measured by the JC-1 staining.The activities of Caspase-9 and Caspase-3 were measured by Caspase assay kit.The expressions of Bcl-2 and Bax mRNA were detected by Real time PCR,and Bcl-2/Bax ratio was analyzed.Results The inhibitory effect of total saponin from allium macrostemon bunge on HeLa cells showed significant time dose response,total saponin from allium macrostemon bunge(100 and 200μg/ml) might significantly decrease the mitochondrial membrane potential of HeLa cells,promote apoptosis (P<0.05,P<0.01,respectively),up regulate the mRNA expression level of Bax,down regulate expression level of Bcl-2 and Bcl-2/Bax ratio,increase the activities of Caspase-9 and Caspase-3 compared with the negative control group(P<0.05,P<0.01,respectively).Conclusion Total saponin from allium macrostemon bunge could effectively inhibit proliferation and induce apoptosis in cervix cancer HeLa cells.The mechanism may be related to decreasing the mitochondrial membrane potential of HeLa cells,up regulating the Bax mRNA expression,down regulating the Bcl-2 mRNA expression,decreasing Bcl-2/Bax ratio and enhancing Caspase-9 and Caspase-3 activities.