白花丹素通过调节线粒体相关旁路蛋白及细胞周期促进非小细胞肺癌H23细胞的凋亡

Plumbagin induces apoptosis via mitochondrial-dependent pathway and cell cycle arrest in human non-small cell lung cancer H23 cells

目的研究天然小分子复合物白花丹素对人类非小细胞癌H23细胞的凋亡作用及其内在机制。方法应用不同浓度梯度的白花丹素（0．5～200μmol／L）培养H23细胞24h，水溶性四唑盐（WST-1）比色法检测H23细胞增殖活力，双染流式细胞仪检测凋亡细胞及细胞周期，计算药物半数抑制浓度（IC50），CM-H2DCFDA探针检测细胞内活性氧化物（ROS），聚丙烯凝胶电泳法检测线粒体旁路相关蛋白。结果白花丹素对于H23细胞系具有极高的抑制增殖的效应，IC50为7．80μmol／L。该抑制增殖效应与G2／M细胞周期停滞，ROS和凋亡相关细胞死亡有关。白花丹素造成细胞周期G2／M的比例上升，并增加ROS的水平。在H23细胞系中，白花丹素增加细胞色素C（Cytochromec）的表达，促凋亡相关蛋白半胱氨酸蛋白酶3（caspase3）、半胱氨酸蛋白酶9（caspase9）的活化，而降低抗凋亡蛋白Bcl-2的表达。结论白花丹素导致H23细胞G2／M周期停滞，并通过线粒体相关旁路诱导人类非小细胞肺癌H23细胞系的凋亡。

Objective This research focused on the effects of plumbagin （5-hydroxy-2-methyl-1,4-naphthoquinone） in the apoptosis of non-small lung cancer cell H23 cells and the underlying mechanisms. Methods H23 cells were treated with different concentration plumbagin （0-200 μmol/L） for 24 hours. The cell growth inhibition activity of plumbagin, IC50, effect on cell cycle distribution, and the effect of plumbagin on mitochondria associated apoptosis and ROS generation were examined. Results Plumbagin exhibited a potent anti-proliferative activity with IC50 values of 7.80 μmol/L in H23 cells. Plumbagin increased the apoptotic rate, G2/M fraction, intracellular level of reactive oxygen species （ROS） in H23 cells in a concentration-dependent manner. The level of activated caspase 3, caspase 9 and Cytochrome C were significantly increased, whereas the level Bcl-2 was decreased in a concentration dependent manner in both cell lines. Conclusion The results indicate that the anti-proliferative effect of plumbagin in H23 cells was associated with G2/M cell arrest, ROS and mitochondrial dependent cell apoptosis pathway.