蛇床子素诱导人肺腺癌细胞进入G2/M阻滞并引起凋亡性死亡研究

Osthole Induces G2/M Arrest and Apoptotic Cell Death in Human Lung Cancer Cells

目的探讨蛇床子素(osthole)对人肺癌细胞A549的杀伤作用及其机制。方法将人肺癌细胞A549用不同浓度的蛇床子素治疗后,采用MTT法检测蛇床子素对A549细胞增殖的抑制作用;通过流式细胞术检测A549细胞内DNA的含量测定蛇床子素对A549细胞周期的影响、蛇床子素处理后A549细胞凋亡情况;Western blot检测蛇床子素处理后A549细胞周期相关蛋白p-Cdc2和Cyclin B1及凋亡相关蛋白Bcl-2和Bax的表达。结果蛇床子素对A549细胞增殖的抑制效应呈剂量依赖性,50μM、100μM、150μM的蛇床子素组增殖抑制率分别为(25.4±3.2)%、(40.2±3.7)%、(63.7±4.5)%。蛇床子素对A549细胞的凋亡诱导效应也呈剂量依赖性,50μM、100μM、150μM的蛇床子素组凋亡率分别为(9.4±1.4)%、(17.3±2.5)%、(22.9±3.3)%。蛇床子素处理后A549细胞周期相关蛋白p-Cdc2、Cyclin B1及抗凋亡蛋白Bcl-2表达显著下降,Bax表达显著增高。结论蛇床子素可诱导人肺腺癌细胞进入G2/M阻滞,并引起肿瘤细胞凋亡性死亡。

Objective To explore the effects of Osthole on the proliferation of A549 cells treated with Osthole and the underlying mechanism. Methods A549 cells were treated with various concentrations of Osthole. Cell proliferation was measured by using the MTT assay. Cell cycle was evaluated by using DNA flow cytometry analysis.Induction of apoptosis was determined by flow cytometry. The expressions of cell cycle related proteins Cyclin B1,p-Cdc2 and apoptosis related proteins Bcl-2, Bax were evaluated by Western blotting. Results Osthole significantly inhibited the proliferation of A549 cells in a dose-dependent manner. The inhibitory rate of A549 cells treated with 50, 100, and 150μmol/L osthole were（25.4±3.2）%,（40.2±3.7）%, and（63.7±4.5）%, respectively. Osthole also increased the apoptosis of A549 cells in a dose-dependent manner. The apoptosis rate of A549 cells treated with50, 100, and 150μmol/L osthole were（9.4 ±1.4）%,（17.3 ±2.5）%, and（22.9 ±3.3）%, respectively. Western blotting showed that Osthole down-regulated the expressions of Cyclin B1, p-Cdc2 and Bcl-2 and up-regulated the expression of Bax in A549 cells. Conclusion Osthole can induce apoptotic cell death and G2/M arrest of A549 cells.