摘要
目的：研究姜黄素作用食管癌的分子机制。方法：体外培养人食管癌细胞（EC109）,15~120μmol/L姜黄素处理后,采用CCK-8法检测姜黄素对细胞的增殖抑制作用,透射电镜观察细胞超微结构,Annexin V-FITC/PI双染激光共聚焦显微镜观察细胞凋亡,流式细胞术分析15~120μmol/L姜黄素作用EC109细胞后PTEN/PI3K/AKT通路相关蛋白PTEN、AKT、GSK3β和Caspase 3的表达水平。结果：CCK-8检测结果姜黄素能显著抑制EC109细胞的增殖,呈剂量和时间效应关系;透射电镜和激光共聚集显微镜观察发现姜黄素能使EC109细胞发生凋亡;流式细胞仪蛋白水平分析显示姜黄素可增强细胞中PTEN、GSK3β和Caspase 3的表达,抑制AKT的表达。结论：姜黄素抑制EC109细胞的增殖并促进其凋亡的生物学效应与增强PTEN的表达抑制PI3K/AKT信号通路有关。
Objective： To study the molecular mechanism of curcumin in human esophageal carcinoma cell line（EC109）. Methods： EC109 cells were cultivated in vitro. When 80%-90% confluence was reached, they were treated with curcumin in different concentrations （15 ~ 120 μmol/L）. The effects on cell proliferation were examined by CCK-8 colorimetry. The ultrastructure of EC109 cells were detected with transmission electron microscope（TEM）. The cells apoptosis was observed with laser confocal microscope（LCM） by AnnexinV-FITC/PI double staining. The proteins level of PTEN, AKT, GSK3β and Caspase 3 were tested by flow cytometry（FCM）. Results： CCK-8 test showed that cureumin could inhibit the proliferation of EC109 cells in a time-and concentration-dependent manner. TEM and LCM examinations indi- cated that curcumin could make ECI09 cells apoptosis. The data of FCM showed that curcumin could increase the expression of FFEN, GSK3β and Caspase 3, decreased the expression of AKT. Conclusion： The effects of curcumin on inhibiting proliferation and promoting apoptosis of EC109 cells were related with increased expression of PFEN and inhibition of PI3K/AKT signaling pathway.
出处
《中国应用生理学杂志》
CAS
CSCD
2015年第2期174-177,共4页
Chinese Journal of Applied Physiology
基金
河北省医学科学研究重点课题计划(ZD20140083)
河北省高等学校科学技术研究青年基金项目(QN20131156)
