發(fā)布時間：2018-05-04 07:05

  本文選題：新藤黃酸 + 阿霉素��； 參考：《中華中醫(yī)藥雜志》2017年06期

【摘要】：目的:研究新藤黃酸(GNA)與阿霉素(ADR)聯(lián)合應(yīng)用對人乳腺癌細胞MCF-7增殖的影響,并進一步探討兩藥聯(lián)用的相關(guān)機制。方法:體外培養(yǎng)人乳腺癌細胞MCF-7細胞株,采用MTT法檢測GNA與ADR單獨及聯(lián)合處理后細胞的存活率;Chou-Talalay聯(lián)合指數(shù)法評價GNA與ADR的藥物聯(lián)合作用;Annexin V-FITC/PI雙染流式細胞儀檢測給藥后MCF-7細胞凋亡率;流式細胞儀檢測JC-1染色細胞線粒體膜電位變化;Western blot法分別檢測GNA與ADR單獨及聯(lián)合處理MCF-7細胞后ASK-1、p-ASK-1、JNK、p-JNK、Cyt-c、Caspase-9和Caspase-3通路蛋白表達的變化,同時檢測經(jīng)JNK通路抑制劑SP600125處理后相關(guān)蛋白指標的變化。結(jié)果:在一定濃度范圍內(nèi),人乳腺癌細胞MCF-7的細胞存活率隨著GNA(0.0625-4.0000μmol/L)與ADR(0.5-16.0μmol/L)給藥劑量增加而降低(P0.01),且聯(lián)合用藥組細胞存活率低于單獨用藥組(P0.01)。聯(lián)合指數(shù)法分析計算后得出合用指數(shù)CI1,提示兩藥聯(lián)合具有協(xié)同作用。GNA與ADR單獨和聯(lián)合作用均能誘導MCF-7細胞凋亡,且聯(lián)合用藥組細胞凋亡率與單獨用藥組比較,顯著增加(P0.01)。單獨給藥組MCF-7細胞線粒體膜電位有所降低(P0.01),而聯(lián)合用藥組與單獨給藥組相比則膜電位明顯降低(P0.01)。與正常對照組比較,單獨給藥組中ASK-1、p-ASK-1、JNK、p-JNK、Cyt-c、Caspase-9和Caspase-3蛋白均被激活,相關(guān)蛋白表達量增加,而聯(lián)合用藥組蛋白表達顯著上調(diào)(P0.01)。使用JNK通路抑制劑SP600125處理后,ASK-1和p-ASK-1蛋白表達與未處理組無明顯差異,Cyt-c和Caspase-9蛋白表達量略微下降,Caspase-3蛋白表達無顯著變化,而SP600125抑制了JNK,p-JNK蛋白表達上調(diào),且聯(lián)合用藥組蛋白表達與單獨給藥組相比有所降低(P0.01)。結(jié)論:GNA與ADR單獨和聯(lián)合應(yīng)用均能誘導人乳腺癌細胞MCF-7細胞凋亡,且兩者聯(lián)合應(yīng)用可產(chǎn)生協(xié)同效應(yīng),其作用機制可能與調(diào)控JNK信號通路相關(guān)。
[Abstract]:Aim: to study the effect of the combination of GNAA and adriamycin on the proliferation of human breast cancer cell line MCF-7, and to explore the mechanism of the combination of the two drugs. Methods: human breast cancer cell line MCF-7 was cultured in vitro. MTT assay was used to detect the survival rate of the cells treated with GNA and ADR alone and in combination with Chou-Talalay index method. The combined effect of GNA and ADR was evaluated by Annexin V-FITC/PI double staining flow cytometry to detect the apoptosis rate of MCF-7 cells. The changes of mitochondrial membrane potential in JC-1 stained cells were detected by flow cytometry and the expression of Caspase-9 and Caspase-3 pathway proteins in MCF-7 cells treated with GNA and ADR alone and in combination with ADR were detected by flow cytometry. At the same time, the changes of related protein indexes were detected after treatment with JNK pathway inhibitor SP600125. Results: within a certain concentration range, the cell survival rate of human breast cancer cell line MCF-7 decreased with the increase of GNA(0.0625-4.0000 渭 mol / L and ADR(0.5-16.0 渭 mol / L administration, and the cell survival rate of the combined treatment group was lower than that of the single drug group. The combined index CI1 was calculated by the combined index method, which indicated that the combination of GNA and ADR could induce apoptosis of MCF-7 cells both alone and in combination, and the apoptosis rate of the combined treatment group was significantly higher than that of the control group. The mitochondrial membrane potential of MCF-7 cells in the single administration group was lower than that in the control group (P 0.01), while the membrane potential in the combination group was significantly lower than that in the control group. Compared with the control group, the expression of Caspase-9 and Caspase-3 in ASK-1 + p-ASK-1 group was activated, and the expression of related proteins was increased, while the expression of histone in combination group was significantly up-regulated (P 0.01). After treatment with JNK pathway inhibitor SP600125, the expression of ASK-1 and p-ASK-1 protein was not significantly different from that of untreated group. The expression of Caspase-3 protein decreased slightly, while SP600125 inhibited the up-regulation of JNK-p-JNK protein expression. The expression of histone in combined drug group was lower than that in control group (P 0.01). Conclusion ADR alone or in combination can induce apoptosis of human breast cancer MCF-7 cells, and the synergistic effect can be produced by the combination of them. The mechanism may be related to the regulation of JNK signaling pathway.
【作者單位】： 安徽中醫(yī)藥大學科研實驗中心省部共建教育部新安醫(yī)學重點實驗中心;
【基金】：國家自然科學基金項目(No.81173600,No.81673650)~~
【分類號】：R737.9
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本文編號：1842071