[关键词]
[摘要]
目的 研究去甲斑蝥素联合马法兰对多发性骨髓瘤细胞增殖与凋亡的影响及其作用机制。方法 去甲斑蝥素单独或联合马法兰给药后,MTT法观察对人多发性骨髓瘤细胞U266株生长的抑制作用,流式细胞术检测细胞凋亡率,Western blotting检测核因子-κB P65(NF-κB P65)、NF-κB抑制因子IκBα、磷酸化IκBα(p-IκBα)、survivin、Bcl-2和Bax蛋白表达的影响。制备多发性骨髓瘤小鼠模型,观察去甲斑蝥素及其与马法兰联合给药对肿瘤生长的抑制作用。结果 去甲斑蝥素能增强马法兰的细胞毒和诱导细胞凋亡作用。与马法兰单独给药组比较,联合给药组使U266细胞核NF-κB P65和胞浆p-IκBα的表达量分别由1.13±0.08、0.83±0.08降至0.26±0.02、0.090±0.002;马法兰对IκBα的表达无影响,但去甲斑蝥素能促进IκBα的表达,二者合用能显著促进胞浆IκBα的表达;与马法兰单独给药组比较,联合给药组survivin和Bcl-2的表达量分别由1.03±0.10、0.72±0.05降至0.52±0.04、0.06±0.01,而Bax由0.29±0.03升至0.75±0.06。体内实验也证实去甲斑蝥素能增强马法兰的抗肿瘤作用。结论 去甲斑蝥素通过抑制NF-κB/p-IκBα途径,调节下游信号分子survivin、Bcl-2和Bax的表达,增强马法兰的抗骨髓瘤作用。
[Key word]
[Abstract]
Abstract: Objective To investigate the effects and mechanisms of norcantharidin (NCTD) combined with melphalan (Mel) on proliferation and apoptosis of multiple myeloma (MM) cells. Methods Human MM cell line U266 cells were treated with NCTD alone or in combination with Mel. MTT and Annexin V/PI flow cytometry were used to determine the rates of cell viability and apoptosis. The protein expression of nuclear factor-κB P65 (NF-κB P65), NF-κB P65 inhibitor IkBα, phosphorylated IκBα (p-IκBα), survivin, Bcl-2, and Bax was determined by Western blotting. The model of mice with MM was established, and the inhibition of combination of NCTD and Mel on tumor growth was observed. Results NCTD potentiated the cytotoxicity and pro-apoptotic effects induced by Mel. Compared with the group treated with Mel alone, the expression levels of NF-κB P65 in U266 nucleus and p-IκBα in cytoplasm in NCTD and Mel combination group decreased from 1.13 ± 0.08 and 0.83 ± 0.08 to 0.26 ± 0.02 and 0.090 ± 0.002, respectively. Mel showed no effect on IκBα expression, but NCTD could enhance its expression, and the combination of NCTD and Mel could obviously enhance the expression. The expression of survivin and Bcl-2 decreased from 1.03 ± 0.10 and 0.72 ± 0.05 to 0.52 ± 0.04 and 0.06 ± 0.01, while the expression of Bax increased from 0.29 ± 0.03 to 0.75 ± 0.06 respectively. In vivo results also demonstrated that NCTD could increase the antitumor effects of Mel. Conclusion The results suggest that NCTD could potentialize the anti-MM effects through inhibiting NF-κB/p-IκBα signal pathway and regulating the espression of survivin, Bcl-2, and Bax.
[中图分类号]
[基金项目]
河北省中医药管理局资助项目(2007136)