二氢青蒿素抑制c-FLIP的表达增强TRAIL对肺癌细胞的凋亡诱导活性

目的: 探讨天然药物二氢青蒿素对TNF相关凋亡诱导配体(TRAIL)抗肺癌活性的影响并研究其机制。方法: 将PC9人肺癌细胞按对照组,二氢青蒿素组,TRAIL组,二氢青蒿素+TRAIL组及二氢青蒿素+TRAIL+c-FLIP质粒组进行分组,MTT法检测二氢青蒿素及TRAIL对PC9细胞的杀伤活性,流式细胞术检测PC9细胞的凋亡和活性氧簇(ROS)的释放,Western blot实验检测PC9细胞c-FLIP的表达、细胞色素c的释放及caspase-8、caspase-9和caspase-3的活化。结果: 二氢青蒿素能显著抑制PC9细胞中c-FLIP蛋白的表达。二氢青蒿素+TRAIL组PC9细胞的相对细胞活力(0.41±0.04)显著低于TRAIL组(0.83±0.06, P<0.05)和二氢青蒿素+TRAIL+c-FLIP质粒组(0.75±0.06, P<0.05)。二氢青蒿素+TRAIL组PC9细胞的凋亡率(35.4±2.6)显著高于TRAIL组(8.7±0.8, P<0.05)和二氢青蒿素+TRAIL+c-FLIP质粒组(13.5±1.1, P<0.05)。二氢青蒿素能显著促进TRAIL诱导的ROS的产生和细胞色素c从线粒体中的释放。二氢青蒿素能显著促进TRAIL诱导的PC9细胞中caspase-8、caspase-9和caspase-3的活化。结论: 二氢青蒿素抑制c-FLIP的表达增强TRAIL对肺癌细胞的凋亡诱导活性。

Dihydroartemisinin enhances TRAIL-induced apoptosis through inhibiting the expression of c-FLIP in lung cancer cells

AIM: To investigate the effect and mechanism of dihydroartemisinin on enhancing the anti-tumor effect of TRAIL on lung cancer. Methods: PC9 cells were divided into control group, dihydroartemisinin group, TRAIL group, dihydroartemisinin + TRAIL group and dihydroartemisinin + TRAIL + c-FLIP plasmid group. MTT assays were performed to evaluate the cytotoxicity of TRAIL and dihydroartemisinin to PC9. Flow cytometry analysis was performed to detect the apoptosis and ROS release in PC9 cells. Western blot analysis was performed to evaluate the expression of c-FLIP, release of cytochrome c and activation of caspase-8, -9 and -3 in PC9 cells. Results: Dihydroartemisinin significantly suppressed the expression of c-FLIP in PC9 cells. Relative cell viability of PC9 in dihydroartemisinin + TRAIL group (0.41±0.04) was significantly lower than the TRAIL group (0.83±0.06, P<0.05) and dihydroartemisinin + TRAIL + c-FLIP group (0.75±0.06, P<0.05). Apoptotic rate of PC9 in dihydroartemisinin + TRAIL group (35.4±2.6) was significantly higher than the TRAIL group (8.7±0.8, P<0.05) and dihydroartemisinin + TRAIL + c-FLIP group (13.5±1.1, P<0.05). Dihydroartemisinin promoted production of ROS and release of cytochrome c from mitochondria. Dihydroartemisinin promoted activation of caspase-8, -9 and -3 in PC9 cells treated with TRAIL. Conclusion: Dihydroartemisinin enhances TRAIL-induced apoptosis through inhibiting the expression of c-FLIP in lung cancer cells.