二氢青蒿素对TNF相关凋亡诱导配体抗肺癌细胞活性的影响及其机制研究

Effect and Mechanism of Dihydroartemisinin on the Activity of TRAIL against Lung Cancer Cells

目的探讨二氢青蒿素对肿瘤坏死因子(TNF)相关凋亡诱导配体(TRAIL)抗肺癌活性的影响及其机制。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表达、细胞色素C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。结论二氢青蒿素通过抑制c-FLIP的表达,增强TRAIL对肺癌细胞的凋亡诱导活性。

Objective To investigate the effect and mechanism of dihydroartemisinin on the cytotoxicity of TNF- related apoptosis-inducing ligand（TRAIL） on lung cancer. Methods PC9 cells were divided into control group, di- hydroartemisinin group, TRAIL group, dihydroartemisinin＋TRAIL group, and dihydroartemisinin＋TRAtL＋e-FLIP plas- mid group. MTT assay was used to evaluate the cytotoxicity of dihydroartemisininand TRAIL to PC9 cells. Flow cytometry was conducted to detect the apoptosis and ROS release in PC9 cells. Western blot was performed to de- termine the expression of c-FLIP, release of cytochrome e, and activation of caspase 8, easpase 9, easpase 3in PC9 ceils. Results Dihydroartemisinin significantly suppressed the expression of c-FLIP in PC9 cells. Relative cell viability of PC9 in dihydroartemisinin ＋ TRAIL group was dramatically lower than that in TRAIL group and dihy- droartemisinin＋TRAIL＋c-FLIP plasmid group[（0.41±0.04） vs （0.83±0.06）, （0.75±0.06）, P〈0.05]. The apoptotie rate of PC9 in dihydroartemisinin ＋TRAIL group was significantly higher than that in TRAIL group and dihy- droartemisinin＋TRAIL＋e-FL1P plasmid group[（35.4±2.6） vs （8.7＋0.8）, （ 13.5±1.1 ）, P〈0.05]. Dihydroartemisinin pro- rooted production of ROS and release of cytochrome c from mitochondria, as well as the activation of caspase 8, caspase 9, caspase 3in PC9 cells treated with TRAIL. Conclusion Dihydroartemisinin enhances TRAIL-induced apoptosis through inhibiting the expression of c-FLIP in lung cancer cells.