【摘要】 目的研究大黄素衍生物A对卵巢癌淋巴结定向高转移细胞SKOV3-pm4的增殖、迁移作用及对Rac1、CDC42基因及蛋白表达的影响。方法实验分为空白对照组、不同浓度药物组(大黄素衍生物A)、Rac1抑制剂组、Rac1激活剂组。以四甲基偶氮唑蓝(MTT)法测定大黄素衍生物A对SKOV3-pm4细胞增殖的抑制作用。免疫荧光法观察SKOV3-pm4细胞中Rac1和CDC42蛋白的分布情况。用细胞划痕实验观察SKOV3-pm4细胞迁移情况,分别采用实时荧光定量PCR(RT-PCR)技术和蛋白免疫印迹法(Western blot)检测细胞中Rac1、CDC42基因和蛋白的表达水平。结果 MTT法检测显示大黄素衍生物A(5~50 mg/L)能抑制SKOV3-pm4细胞的增殖,24 h的半数抑制浓度(IC50)为34.46 mg/L。细胞划痕实验显示4 mg/L的大黄素衍生物A能抑制SKOV3-pm4细胞迁移,且呈时间依赖性。免疫荧光观察到细胞中Rac1和CDC42蛋白均广泛分布于细胞质中,Rac1蛋白在细胞核周围较为密集。RT-PCR和Western blot法检测结果显示随着大黄素衍生物A药物浓度的梯度增加,Rac1基因和蛋白的表达水平均降低,与空白对照组比较,差异均有统计学意义(P均<0.05),而CDC42基因和蛋白的表达变化不明显。结论大黄素衍生物A能抑制SKOV3-pm4细胞增殖和迁移能力,其作用途径可能是通过调控Rac1信号传导通路而实现;Rac1可能是大黄素衍生物A潜在的作用靶点。更多还原

【Abstract】 Objective To study the effects of Emodin Derivative A on SKOV3-pm4 ovarian cancer cells with lymph node-oriented metastasis,particularly on expression of Rac1 and CDC42 m RNA and protein. Methods SKOV3-pm4 cells were treated for 24 h with different concentrations of Emodin Derivative A in the presence or absence of Rac1 inhibitor or activator. Cell proliferation was assessed by MTT assay,cell migration,by scratch assay;Rac1 and CDC42 expression,by RT-PCR and Western blotting;and distribution of these two proteins,by immunofluorescence. Results Emodin Derivative A at doses of 5-50 mg/L inhibits the proliferation of SKOV3-pm4 cells(IC5034.46 mg/L). Rac1 and CDC42 protein were present throughout the cytoplasm,though Rac1 staining was denser around the nucleus than in other parts of the cell. Expression of Rac1 m RNA and protein decreased as the dose of Emodin Derivative A increased. CDC42 expression,however,did not change significantly with drug dose. Emodin Derivative A at a dose of 4 mg/L inhibited cell migration. Conclusions Emodin Derivative A can inhibit migration and proliferation of SKOV3-pm4 cells,and the drug may act by targeting Rac1.更多还原