灯盏乙素通过下调TRIM32的表达增强卡铂的抗卵巢癌活性研究

Scutellarin Enhances Carboplatin-induced Cytotoxicity Against Ovarian Cancer by Decrease the Expression of TRIM32

关键词:  灯盏乙素  TRIM32  卡铂  卵巢癌

Key words:  scutellarin  TRIM32  carboplatin  ovarian cancer

摘要:

目的 探讨灯盏乙素对卵巢癌卡铂化疗的辅助治疗作用并研究其机制。方法 CCK-8试验检测卵巢癌细胞系A2780在灯盏乙素和不同浓度卡铂处理下的细胞活力。Western blotting试验检测卡铂和灯盏乙素对A2780细胞TRIM32、Bcl-2及caspase-9、caspase-3活化水平的影响。流式细胞术检测A2780细胞活性氧簇（reactive oxygen species，ROS）的水平、线粒体膜电位和细胞凋亡率。结果 灯盏乙素联合处理能明显降低卡铂对A2780细胞的IC₅₀。灯盏乙素处理能显著抑制A2780细胞TRIM32的表达。转染TRIM32真核表达质粒后，灯盏乙素对卡铂抗卵巢癌活性的增强作用受到明显抑制。灯盏乙素处理能明显降低A2780细胞Bcl-2的表达并且促进卡铂诱导ROS产生。灯盏乙素联合卡铂能引起显著的线粒体膜电位的降低，caspase-9、caspase-3的活化及凋亡的发生。转染TRIM32真核表达质粒后，灯盏乙素联合卡铂对A2780细胞线粒体途径的凋亡受到显著抑制。结论 灯盏乙素通过下调TRIM32的表达增强卡铂对卵巢癌细胞线粒体途径凋亡的诱导作用。

Abstract:

OBJECTIVE To investigate the adjuvant effect and mechanisms of scutellarin in carboplatin-based chemotherapy on ovarian cancer. METHODS CCK-8 assay was performed to evaluate the effect of scutellarin and different concentrations of carboplatin on changing the viability of A2780 ovarian cancer cells. Western blotting analysis was performed to detect the effect of scutellarin and carboplatin on changing the expression of TRIM32 and Bcl-2 and activation of caspase-9 and caspase-3 in A2780 cells. Flow cytometry analysis was performed to detect the reactive oxygen species(ROS) level, mitochondrial membrane potential and apoptotic rate of A2780 cells. RESULTS Adjuvant therapy of scutellarin significantly decreased the IC₅₀ of carboplatin to A2780 cells. Scutellarin obviously inhibited the expression of TRIM32 in A2780 cells. However, transfection with TRIM32 plasmid suppressed the effect of scutellarin on enhancing the cytotoxicity of carboplatin against ovarian cancer. Scutellarin decreased the expression of Bcl-2 and promoted the carboplatin-induced generation of ROS. Combination with carboplatin and scutellarin significantly induced the collapse of mitochondrial membrane potential, activation of caspase-9 and caspase-3, and occurrence of cell apoptosis in A2780 cells. However, transfection with TRIM32 plasmid obviously suppressed the mitochondrial apoptosis pathway induced by the combination treatment with scutellarin and carboplatin in the A2780 cells. CONCLUSION Scutellarin decrease the expression of TRIM32 and thus enhance the carboplatin-induced mitochondrial apoptosis in ovarian cancer.