斑蝥素酸镁阻断MAPK信号通路抑制SMMC-7721人肝癌细胞增殖

发布时间：2018-07-18 11:57

【摘要】：目的观察斑蝥素酸镁对SMMC-7721肝癌细胞丝裂原激活蛋白激酶(MAPK)信号通路的影响,探讨斑蝥素酸镁的抗癌机制。方法使用蛋白磷酸酶2A(PP2A)活性检测试剂盒分别检测斑蝥素酸镁和冈田酸(OA)对PP2A活性的影响。实时定量PCR检测斑蝥素酸镁、OA对SMMC-7721人肝癌细胞胞外信号调节激酶1/2(ERK1/2)、p38MAPK、c-Jun N末端激酶1/2(JNK1/2)mRNA表达水平的影响;Western blot法检测斑蝥素酸镁、OA对SMMC-7721细胞ERK1/2、p38MAPK、JNK蛋白表达以及蛋白磷酸化水平的影响。结果 0.283μmol/L斑蝥素酸镁对PP2A活性无明显抑制作用,0.567μmol/L斑蝥素酸镁能显著抑制PP2A活性,且随着药物浓度的增加,抑制作用愈趋明显;同时0.059 nmol/L OA对PP2A活性也有显著抑制作用。与空白对照组比较,0.283μmol/L斑蝥素酸镁组ERK1、ERK2 mRNA表达量无明显变化,当浓度为0.567μmol/L时,ERK1、ERK2mRNA表达量显著下降,且随着药物浓度的增加下降更明显;而0.059 nmol/L OA组ERK1、ERK2 mRNA表达量却显著升高。0.059 nmol/L OA和不同浓度斑蝥素酸镁组的p38MAPK、JNK1、JNK2 mRNA表达量均显著升高。与空白对照组比较,0.283μmol/L斑蝥素酸镁组ERK1/2磷酸化水平无明显变化,高于0.567μmol/L斑蝥素酸镁处理显著下调ERK1/2磷酸化水平,其下调程度具有浓度依赖效应;而0.059 nmol/L OA组ERK1/2磷酸化水平却显著上调。0.059 nmol/L OA和不同浓度斑蝥素酸镁组的p38MAPK、JNK磷酸化水平均显著上调。结论斑蝥素酸镁可能是通过抑制PP2A活性进而抑制ERK1/2通路来实现对SMMC-7721肝癌细胞增殖的抑制作用。
[Abstract]:Objective to investigate the effect of magnesium cantharidin on mitogen-activated protein kinase (MAPK) signaling pathway in SMMC-7721 hepatoma cell line and to explore the anticancer mechanism of magnesium cantharidin. Methods protein phosphatase 2A (PP2A) activity assay kit was used to detect the effects of Cantharidin magnesium and Okadaic acid (OA) on PP2A activity. Effects of Cantharidin magnesium Oligonidate on the expression of extracellular signal-regulated kinase 1 / 2 (ERK1 / 2) p38MAPKnc-Jun N-terminal kinase 1 / 2 (JNK1 / 2) mRNA in SMMC-7721 human hepatocarcinoma cell line SMMC-7721 by real-time quantitative PCR. The effects of cantharidin magnesium aspartate OA on the expression of ERK1 / 2p38MAPKNK JNK protein and protein phosphorylation in SMMC-7721 cells were detected by Western blot. Results magnesium cantharidate (0.283 渭 mol / L) had no obvious inhibitory effect on PP2A activity. Magnesium cantharidate (0.567 渭 mol / L) could significantly inhibit PP2A activity, and the inhibitory effect increased with the increase of drug concentration, and 0.059 nmol / L OA had a significant inhibitory effect on PP2A activity. Compared with the control group, the expression of ERK1 ERK2 mRNA in 0.283 渭 mol / L magnesium cantharidate group had no significant change, and the expression of ERK1 ERK2 mRNA decreased significantly when the concentration was 0.567 渭 mol / L, and the decrease was more obvious with the increase of drug concentration. However, the expression of ERK1 + ERK2 mRNA in 0.059 nmol / L OA group was significantly higher than that in the control group. The expression of p38 MAPK1 JNK1 JNK2 mRNA was significantly increased in the 0.059 nmol / L OA group and in the different concentration of magnesium cantharidate group. Compared with the blank control group, the phosphorylation level of ERK1 / 2 in the 0.283 渭 mol / L magnesium cantharidate group did not change significantly, but was significantly lower than that in the 0.567 渭 mol / L magnesium cantharidate group, and the down-regulation was concentration-dependent. However, the phosphorylation level of ERK1 / 2 in 0.059 nmol / L OA group was significantly higher than that in 0.059 nmol / L OA group and p38 MAPKN JNK phosphorylation level in different concentration magnesium cantharidate group. Conclusion magnesium cantharidate may inhibit the proliferation of SMMC-7721 hepatoma cells by inhibiting PP2A activity and ERK1 / 2 pathway.
【作者单位】：遵义医学院贵州省普通高等学校特色药物肿瘤防治特色重点实验室;遵义医学院基础医学院;
【基金】：国家自然科学基金(81560669,81260488) 贵州省科技合作计划项目(黔科合LH字[2015]7509号,黔科合LH字[2015]7516号) 贵州省科技厅中药现代化项目(黔科合ZY字[2013]3012) 贵州省卫计委科学基金项目(gzwjk2015-1-013) 医学昆虫资源开发利用创新团队(黔合教人才团队字[2014]39号) 贵州省教育厅特色重点实验室建设项目(黔教合KY字[2014]212)
【分类号】：R735.7

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