木犀草苷对人食管鳞状癌Eca109细胞生长的抑制作用及其机制

Inhibition of luteoloside on proliferation of human esophageal carcinoma cell line Eca109 and its mechanism

目的研究木犀草苷对人食管鳞状癌Eca109细胞生长的抑制作用及其机制。方法 Eca109细胞经不同浓度木犀草苷处理后,MTT法检测Eca109细胞增殖;倒置显微镜观察Eca109细胞形态的变化;流式细胞术检测Eca109细胞周期变化及细胞凋亡情况;RT-PCR检测Eca109细胞cyclin D1、survivin和c-myc基因表达。结果 MTT实验表明,木犀草苷80、120、160、200、240μmol/L对Eca109细胞均有抑制作用,且与浓度相关。木犀草苷可引起Eca109细胞形态改变、体积缩小,并与周围细胞脱离,浓度为240μmol/L时使细胞呈出芽状,有的细胞伸出多个伪足样突起;浓度为160、240μmol/L时,可将细胞阻滞于G2/M期,诱导细胞凋亡;浓度为240μmol/L并处理Eca109细胞48 h后,使Eca109细胞cyclin D1、survivin和c-myc基因表达低于对照组(P<0.05)。结论木犀草苷对食管鳞状癌Eca109细胞的生长有显著抑制作用,其通过改变细胞周期、诱导细胞凋亡及抑制相关基因的表达发挥抗肿瘤作用。

Objective To investigate the inhibition of luteoloside on the proliferation of human esophageal carcinoma cell line Eca109 and its mechanism.Methods MTT assay was used for detecting the influence of luteoloside at different concentration on the proliferation of Eca109.The morphological changes of cells were observed under inverted microscope.The cell cycle and apoptosis were detected by flow cytometry.RT-PCR was used to detect the mRNA expression of cyclin D1,survivin,and c-myc genes.Results The results of MTT assay showed different doses（80,120,160,200,and 240 μmol/L） of luteoloside could inhibit the cell proliferation of Eca109 cells in a dose-response manner.Luteoloside could also change the morphological characteristics of cells,reduce the cell size,and separate from peripheral cells.Treated with 240 μmol/L luteoloside,the cells were sprouting and some of them developed multiple pseudopodia-like protrusions.Treated with luteoloside（160 and 240 μmol/L）,the cell cycle of Eca109 cells was blocked in G2/M phase and apoptosis was induced（P〈 0.05）.Compared with the control group,the gene expression of cyclin D1,survivin,and c-myc was decreased after treated with 240 μmol/L luteoloside for 48 h（P〈0.05）.Conclusion Luteoloside could significantly inhibit the proliferation of Eca109 cells by changing the cell cycle and inducing the apoptosis.Moreover,it could decrease the mRNA expression of relative genes.