槐定碱下调FoxM1在食管胃交界腺癌中的增殖、凋亡研究

Sophoridine Inhibits Proliferation and Induces Apoptosis Through Downregulation of FoxM1 in Human Adenocarcinoma from Esophagogastric Junction

目的分析槐定碱抑制食管胃交界腺癌细胞系增殖及诱导凋亡的分子机制。方法采用0~3.0 mg·mL^(-1)质量浓度处理OE^(-1)9和SK-GT2细胞24、48和72 h,分别用CCK-8检测细胞增殖、流式细胞计数检测凋亡和细胞周期、生化分析检测细胞内活性氧(ROS)和谷胱甘肽(GSH)含量、Real-time PCR和Western blot检测FoxM1表达、双荧光素酶报告基因实验检测FoxM1启动子转录活性。结果槐定碱明显抑制食管胃交界腺癌细胞增殖,诱导细胞凋亡,阻滞细胞在G0/G1期,诱导OE^(-1)9细胞内ROS的产生并同时使细胞内GSH含量的减少,其有效作用质量浓度为0.5~1.0 mg·mL^(-1)。此外,槐定碱能够通过抑制FoxM1启动子转录活性而下调FoxM1的mRNA和蛋白表达。结论槐定碱能够通过下调FoxM1基因的表达抑制食管胃交界腺癌细胞的体外增殖。

OBJECTIVE To investigate the molecular mechanisms of proliferation inhibition and apoptosis induction by sophoridine on cell lines from adenocarcinoma in esophagogastric junction.METHODS After treatment of OE-19 and SK-GT2 cells with 0-3.0 mg·mL^-1 of sophoridine for 24,48 and 72 h,CCK8 was used to examine the proliferation,flow cytometry was used to examine apoptosis,biochemical assay for intracellular ROS and GSH,real-time PCR and Western blot were used to examine FoxM1 mRNA and protein expression,and dual-luciferase reporter gene assay was used for measurement of the transcriptional activity of FoxM1.RESULTS Sophoridine could significantly inhibit the proliferation of OE-19 and SK-GT2 cell lines,induces apoptosis and G0/G1 arrest of OE-19 cell lines at the concentration of 0.5-1.0 mg·mL^-1.Intracellular ROS increase and GSH decrease were observed as well.Moreover,sophoridine attenuated the expression of FoxM1 through suppression of its transcriptional activity.CONCLUSION It suggests that sophoridine may inhibit proliferation and induce apoptosis of adenocarcinoma from esophagogastric junction in vitro through down-regulating the expression of FoxM1.