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鉴于白花丹素(Plumbago zeylanica L.)抗癌谱广、毒性低的特点,分析白花丹素抑制食管鳞癌细胞系增殖及诱导凋亡的分子机制对其进一步的结构改造和临床应用具有重要的理论意义。本研究采用0~20μmol·L~(-1)浓度处理KYSE-30、KYSE-70和KYSE~(-1)40细胞24、48和72 h,分别用CCK-8检测细胞增殖、Annexin V和PI荧光染色检测凋亡、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。并采用裸鼠体内治疗实验分析白花丹素抗肿瘤作用与FoxM1的关系。结果显示,白花丹素明显抑制食管鳞癌细胞增殖,诱导细胞凋亡,抑制细胞在体内的增殖,其有效作用浓度为5~10μmol·L~(-1)。此外,研究发现白花丹素能够通过抑制FoxM1启动子转录活性而下调FoxM1的m RNA和蛋白表达。本研究表明,白花丹素能够通过下调FoxM1基因的表达抑制食管癌细胞的体内体外增殖。
In view of the broad spectrum of anticancer activity and low toxicity of plumbago zeylanica L., the molecular mechanism of plumbago zingiberis inhibiting the proliferation and inducing apoptosis of esophageal squamous carcinoma cell lines is of great importance to its further structural transformation and clinical application Theoretical significance. In this study, KYSE-30, KYSE-70 and KYSE ~ (40) 40 cells were treated with 0 ~ 20μmol·L -1 for 24,48 and 72 h respectively. Cell proliferation, Annexin V and PI staining was used to detect apoptosis. FoxM1 expression was detected by real-time PCR and Western blot. The transcriptional activity of FoxM1 promoter was detected by dual luciferase reporter assay. And in nude mice in vivo treatment experiment analysis plumbagin antitumor effect and FoxM1 relationship. The results showed that plumbagin significantly inhibited the proliferation of esophageal squamous cell carcinoma cells, induced apoptosis and inhibited cell proliferation in vivo. The effective concentration of plumbagin was 5 ~ 10μmol·L -1. In addition, the study found that plumbagin can down-regulate FoxM1 m RNA and protein expression by inhibiting the FoxM1 promoter transcriptional activity. This study showed that plumbagin can inhibit the proliferation of esophageal cancer cells in vitro and in vivo by down-regulating FoxM1 gene expression.