松果菊苷对人肺癌HCC827细胞增殖、凋亡和迁移的影响

Effect of Echinacoside on Proliferation,Apoptosis and Migration of Human Lung Cancer Cell Line HCC827

目的探讨松果菊苷对人肺癌HCC827细胞增殖、凋亡和迁移的影响。方法采用MTT法、以酶标仪分别检测松果菊苷(0,5,10,20,40,80,160μmol/L)和顺铂(0,5,10,20,40,80,160,320μmol/L)条件下HCC827细胞的生长情况,并计算细胞增殖率及半数致死浓度(LC_(50));实验分为阴性对照组(等体积培养基)、阳性对照组(顺铂80μmol/L)及松果菊苷低、中、高剂量组(实验Ⅰ、Ⅱ、Ⅲ组,20,40,80μmol/L)。采用细胞划痕法检测细胞迁移能力,流式细胞术检测细胞凋亡情况,实时荧光定量聚合酶链式反应法及Western blot法分别检测细胞中转化生长因子βⅠ型受体(TβRⅠ)、Smad 2和Smad 3 mRNA及蛋白表达水平。结果松果菊苷和顺铂的LC_(50)分别为(47.50±6.21)μmol/L和(76.85±14.89)μmol/L。与阴性对照组比较,阳性对照组和实验Ⅰ、Ⅱ、Ⅲ组的细胞迁移距离均显著缩短,细胞凋亡率均显著升高,TβRⅠ、Smad 2和Smad 3 mRNA及蛋白表达水平均显著降低(P<0.05)。结论松果菊苷可抑制HCC827细胞增殖及迁移,并促进细胞凋亡,机制可能与抑制TβRⅠ/Smad信号通路激活有关。

Objective To investigate the effect of echinacoside on proliferation,apoptosis and migration of human lung cancer cell line HCC827.Methods Methyl thiazolyl tetrazolium(MTT)assay and microplate reader were used to measure the growth of HCC827 cells under the conditions of echinacoside(0,5,10,20,40,80,160μmol/L)and cisplatin(0,5,10,20,40,80,160,320μmol/L),and the cell proliferation rate and half inhibitory concentration(LC_(50))were calculated.The experiment was divided into the negative control group(equal volume of medium)and the positive control group(80μmol/L of cisplatin)and echinacoside low-,medium-and high-dose groups(experimental Ⅰ,Ⅱ,Ⅲ groups,20,40,80μmol/L).The cell migration ability was detected by the cell scratch method,the cell apoptosis rate was detected by the flow cytometry,and the mRNA and protein expression levels of transforming growth factor typeβⅠreceptor(TβRⅠ),Smad 2 and Smad 3 were detected by the real-time fluorescence quantitative polymerase chain reaction(RT-PCR)and Western blot respectively.Results The LC50 of echinacoside and cisplatin were(47.50±6.21)μmol/L and(76.85±14.89)μmol/L,respectively.Compared with those in the negative control group,the distance of cell migration was significantly shorter,and the cell apoptosis rate was significantly higher,while the mRNA and protein expression levels of TβRⅠ,Smad 2 and Smad 3 were significantly lower in the positive control group and experimental Ⅰ,Ⅱ,Ⅲ groups(P<0.05).Conclusion Echinacoside can inhibit the proliferation and migration of HCC827 cells,and promote cell apoptosis,and the mechanism may be related to the inhibition of echinacoside on the activation of TβRⅠ/Smad signaling pathway.