發(fā)布時間：2018-07-18 11:57

【摘要】：目的觀察斑蝥素酸鎂對SMMC-7721肝癌細胞絲裂原激活蛋白激酶(MAPK)信號通路的影響,探討斑蝥素酸鎂的抗癌機制。方法使用蛋白磷酸酶2A(PP2A)活性檢測試劑盒分別檢測斑蝥素酸鎂和岡田酸(OA)對PP2A活性的影響。實時定量PCR檢測斑蝥素酸鎂、OA對SMMC-7721人肝癌細胞胞外信號調(diào)節(jié)激酶1/2(ERK1/2)、p38MAPK、c-Jun N末端激酶1/2(JNK1/2)mRNA表達水平的影響;Western blot法檢測斑蝥素酸鎂、OA對SMMC-7721細胞ERK1/2、p38MAPK、JNK蛋白表達以及蛋白磷酸化水平的影響。結(jié)果 0.283μmol/L斑蝥素酸鎂對PP2A活性無明顯抑制作用,0.567μmol/L斑蝥素酸鎂能顯著抑制PP2A活性,且隨著藥物濃度的增加,抑制作用愈趨明顯;同時0.059 nmol/L OA對PP2A活性也有顯著抑制作用。與空白對照組比較,0.283μmol/L斑蝥素酸鎂組ERK1、ERK2 mRNA表達量無明顯變化,當濃度為0.567μmol/L時,ERK1、ERK2mRNA表達量顯著下降,且隨著藥物濃度的增加下降更明顯;而0.059 nmol/L OA組ERK1、ERK2 mRNA表達量卻顯著升高。0.059 nmol/L OA和不同濃度斑蝥素酸鎂組的p38MAPK、JNK1、JNK2 mRNA表達量均顯著升高。與空白對照組比較,0.283μmol/L斑蝥素酸鎂組ERK1/2磷酸化水平無明顯變化,高于0.567μmol/L斑蝥素酸鎂處理顯著下調(diào)ERK1/2磷酸化水平,其下調(diào)程度具有濃度依賴效應;而0.059 nmol/L OA組ERK1/2磷酸化水平卻顯著上調(diào)。0.059 nmol/L OA和不同濃度斑蝥素酸鎂組的p38MAPK、JNK磷酸化水平均顯著上調(diào)。結(jié)論斑蝥素酸鎂可能是通過抑制PP2A活性進而抑制ERK1/2通路來實現(xiàn)對SMMC-7721肝癌細胞增殖的抑制作用。
[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.
【作者單位】： 遵義醫(yī)學院貴州省普通高等學校特色藥物腫瘤防治特色重點實驗室;遵義醫(yī)學院基礎(chǔ)醫(yī)學院;
【基金】：國家自然科學基金(81560669,81260488) 貴州省科技合作計劃項目(黔科合LH字[2015]7509號,黔科合LH字[2015]7516號) 貴州省科技廳中藥現(xiàn)代化項目(黔科合ZY字[2013]3012) 貴州省衛(wèi)計委科學基金項目(gzwjk2015-1-013) 醫(yī)學昆蟲資源開發(fā)利用創(chuàng)新團隊(黔合教人才團隊字[2014]39號) 貴州省教育廳特色重點實驗室建設(shè)項目(黔教合KY字[2014]212)
【分類號】：R735.7

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