半枝莲提取物对人肺癌A549细胞的凋亡作用及其对凋亡相关基因表达的影响

Apoptotic effect of scutellaria barbata D.Don extract on human lung cancer cell A549 and effect on expression of apoptosis related genes

目的:了解半枝莲提取物对人肺癌A549细胞的凋亡作用及其对凋亡相关基因表达的影响,探讨半枝莲提取物的抗肺癌作用及其分子生物学作用机制。方法:采用70%乙醇水溶液制备半枝莲提取物;实验组的半枝莲提取物浓度为3mg/L、6mg/L、12mg/L,对照组不加药物。采用MTT法测定药物对细胞生长的抑制率;采用流式细胞仪检测细胞凋亡率;采用RT-PCR法测定细胞凋亡相关基因Survivin、Caspase-3和内对照β-actin的表达水平。结果:3mg/L、6mg/L、12mg/L浓度实验组的OD值均低于对照组;实验组的抑制率分别为(23.53±5.84)%、(39.71±6.77)%和(66.18±8.45)%。对照组3mg/L、6mg/L、12mg/L浓度实验组的细胞凋亡率分别是(3.76±0.82)%、(16.61±3.19)%、(21.34±4.52)%、(35.19±5.80)%;3mg/L、6mg/L、12mg/L浓度实验组的细胞凋亡率均高于对照组。3mg/L、6mg/L、12mg/L浓度实验组的Survivin/β-actin均低于对照组;3mg/L、6mg/L、12mg/L浓度实验组的Caspase-3/β-actin均高于对照组。结论:半枝莲提取物可通过诱导细胞凋亡作用抑制人肺癌A549细胞的生长,其分子机制为下调Survivin的表达水平和上调Caspase-3的表达水平。

Objective：To learn the apoptotic effect of Scutellaria barbata D.Don extract on human lung cancer cell A549 and effect on the expression of apoptosis related genes.To investigate the anti lung cancer effect of Scutellaria barbata D.Don extract and its molecular mechanism.Methods：Scutellaria barbata D.Don extract was prepared by 70%ethanol aqueous solution.The concentrations of Scutellaria barbata D.Don extract in experimental groups were 3mg/L,6mg/L and 12mg/L.The control group without drug.MTT method was used to determine the inhibitory rate of drug on cell growth.The apoptosis rate was detected by flow cytometry.The expression levels of apoptosis related genes Survivin,Caspase-3and internal control geneβ-actin were determined by RT-PCR method.Results：ODs of 3mg/L,6mg/L and 12mg/L concentration experimental groups were lower than the control group;inhibition rates of the experimental groups were（23.53±5.84）% and（39.71±6.77）% and（66.18±8.45）%,respectively.The apoptosis rates of the control group,3mg/L,6mg/L,12mg/L concentration experimental groups were（3.76±0.82）%,（16.61±3.19）%,（21.34 ±4.52）%,（35.19±5.80）%;3mg/L,6mg/L,12mg/L concentration experimental groups were higher than the control group.Survivin/β-actin ratios of 3mg/L,6mg/L,12mg/L concentration experimental group were lower than the control group.Caspase-3/β-actin ratios of 3mg/L,6mg/L,12mg/L concentration experimental group were higher than the control group.Conclusions：Scutellaria barbata D.Don extract can inhibit the growth of human lung cancer A549 cells by inducing cell apoptosis,and the molecular mechanism are down-regulating the expression level of Survivin and up-regulating expression level of Caspase-3.