摘要
目的探讨苦参碱(Mat)对视网膜母细胞瘤细胞Y79细胞增殖、凋亡及PI3K/Akt信号通路的影响。方法Y79细胞经0.5、1.0、1.5 g/L Mat处理24 h后,采用CCK-8试剂盒检测细胞增殖情况,磷酯酰丝氨酸结合蛋白-异硫氢酸荧光素/碘化丙啶(Annexin V-FITC/PI)双标记流式细胞术检测细胞凋亡情况,Western blotting免疫印迹检测各浓度Mat处理后凋亡相关蛋白Bcl-2及Bax、PI3K/Akt信号通路重要蛋白PI3K p85亚基、Akt及其磷酸化形式p-Akt的蛋白水平。结果不同浓度Mat作用Y79细胞24 h后,可呈浓度依赖性方式抑制细胞增殖,增加细胞凋亡率(P<0.05)。Western blotting检测发现,Mat可明显增加促凋亡蛋白Bax表达,降低抗凋亡蛋白Bcl-2水平并可降低PI3K p85α亚基及磷酸化Akt的蛋白水平(P<0.05),对总Akt水平无明显影响。结论 Mat可抑制人视网膜母细胞瘤增殖并诱导其凋亡,可能与抑制PI3K/Akt信号通路有关。
Objective To investigate the effect of matrine （Mat） on the proliferation, apoptosis and PI3K/Akt pathway of ret- inoblastoma（Rib） ceils. Methods Rb cell lines Y79 ceils were cultured with different concentration of Mat（0. 5, 1. 0, 1. 5g/L）. Cell proliferation and apoptosis were determined by CCK-8 assay and flow cytometry（ stained with Annexin V-FITC/PI）. Apoptosis related genes Bax and Bcl-2, as well as the expression of phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） pathways protein, inclu- ding p85ct, p-Aktset473 and Akt, were determined by Western blotting. Results Mat could inhibit cell proliferation and induce apoptosis in a dose-dependent manner, respectively. Moreover, Mat increased the expression of Bax, but decreased the Bcl-2 protein expression. Results of Western blotting showed that Mat decreased the level of PI3K p85ct and phosphor-Akt. Conclusion Mat could inhibit the proliferation and induce the apoptosis of Rb ceils, probably modulating by the PI3K/Akt pathway.
出处
《临床肿瘤学杂志》
CAS
2015年第7期598-601,共4页
Chinese Clinical Oncology
