欧前胡素通过上调细胞表面死亡受体5的数量增强TRAIL对乳腺癌细胞的杀伤活性

目的:探讨中药活性成分欧前胡素对肿瘤坏死因子相关凋亡诱导配体(TRAIL)的协同抗乳腺癌效应及分子机制。方法:将人乳腺癌细胞T-47D和MCF-7按对照组、欧前胡素组、TRAIL组、欧前胡素+TRAIL组及欧前胡素+TRAIL+死亡受体5(DR5)siRNA组进行分组,MTT法检测T-47D和MCF-7细胞活力,流式细胞术检测T-47D细胞凋亡和线粒体膜电位,Western blot实验和流式细胞术检测T-47D细胞表面DR5的表达水平及caspase-8、caspase-3的活化水平。结果:MTT实验结果显示,欧前胡素联合用药能显著提高各浓度TRAIL对T-47D和MCF-7细胞的杀伤活性;流式细胞术和Western blot结果显示欧前胡素处理能显著提高T-47D细胞DR5的表达水平和活性氧簇产生水平(P0.05)。另外,流式细胞术和Western blot结果还显示,欧前胡素联合用药能显著增强TRAIL促进T-47D细胞线粒体膜电位损伤、caspase-8和caspase-3活化及凋亡的作用。结论:欧前胡素通过上调乳腺癌细胞DR5的表达水平发挥对TRAIL的协同抗乳腺癌效应。

Imperatorin enhances anti-tumor effect of TRAIL on breast cancer by upregulating expression of DR5 on cell surface

AIM: To investigate the synergistic effect of imperatorin on enhancing the anti-tumor effect of TNF-related apoptosis-inducing ligand (TRAIL) on breast cancer and the mechanisms. METHODS: T-47D and MCF-7 breast cancer cells were divided into control group, imperatorin group, TRAIL group, imperatorin+TRAIL group and imperatorin+TRAIL+death receptor 5 (DR5) siRNA group. The viability of T-47D and MCF-7 cells was measured by MTT assay. The apoptosis and mitochondrial membrane potential in T-47D cells were analyzed by flow cytometry. Western blot and flow cytometry analysis were performed to evaluate the expression of DR5 on T-47D cell surface and the activation of caspase-8 and caspase-3. RESULTS: Imperatorin significantly enhanced the inhibition of cell viability induced by TRAIL of T-47D and MCF-7 cells, and significantly increased the apoptosis of T-47D cells induced by TRAIL. Imperatorin treatment ob-viously induced upregulation of DR5 expression and production of reactive oxygen species in the T-47D cells. In addition, imperatorin enhanced the TRAIL-induced damage of mitochondrial membrane potential and activation of caspase-8 and caspase-3. CONCLUSION: Imperatorin enhances the anti-tumor effect of TRAIL on breast cancer via upregulating the expression of DR5.