香加皮宝霍甙-Ⅰ抑制人食管癌细胞增殖的机制

Mechanism of Baohuoside-Ⅰ from Cortex Periplocae Inhibits Cell Proliferation of Human Esophageal Carcinoma

目的观察香加皮活性成分宝藿甙-Ⅰ对食管癌细胞增殖活性及凋亡的影响,并探讨其与β-catenin通路的关系。方法采用MTT法观察宝藿甙-Ⅰ对食管癌细胞Eca-109增殖活性的影响;An-nexin V/PI染色检测宝藿甙-Ⅰ对食管癌细胞凋亡的影响。RT-PCR法检测宝藿甙-Ⅰ对食管癌细胞β-catenin、survivin和c-myc mRNA表达的影响,Western blot和激光共聚焦显微镜检测食管癌细胞β-catenin、磷酸化β-catenin、survivin和c-myc蛋白表达的变化。结果 10～40μg/ml宝藿甙-Ⅰ明显降低了Eca-109细胞的增殖活性并诱导其凋亡。经宝藿甙-Ⅰ处理24 h后,β-catenin、survivin和c-myc mR-NA和蛋白表达水平均降低,磷酸化的β-catenin蛋白表达升高,β-catenin在细胞核和细胞质中的表达均降低,且在胞核中的降低更为明显。结论香加皮宝藿甙-Ⅰ可通过调节β-catenin通路活性抑制食管癌Eca-109细胞增殖。

Objective Inhibition of baohuoside-Ⅰ from Cortex Periplocae（CP） on proliferation of human esophageal carcinoma cells was investigated.The relationship between apoptosis and beta-catenin pathway also was explored.Methods The proliferation of Eca109 cells was measured by MTT assay.Annexin V/PI staining was used to detect the apoptotic rate of Eca-109 cells.Effect of baohuoside-Ⅰ on level of β-catenin,survivin and c-myc mRNA were detected by RT-PCR.Expression of β-catenin,phosphorylate β-catenin,survivin and c-myc protein were determined by western blot and TEM.Results It is demonstrated that baohuoside-Ⅰ could inhibit the proliferate activity of Eca-109 cells by inducing apoptosis.After treatment with baohuoside-Ⅰ for 24 h,β-catenin,survivin and c-myc mRNA expression in Eca-109 decreased obviously.Moreover,baohuoside-Ⅰ reduced the protein levels of β-catenin,phosphorylate β-catenin,survivin and c-myc in Eca-109 cells.Conclusion Baohuoside-Ⅰ isolated from cortex periplocae inhibited proliferation of esophageal carcinoma cells through β-catenin signal pathway.