中药陵水暗罗提取物暗罗素逆转卵巢癌细胞顺铂耐药研究

Zincpolyanemine from Chinese Herbs Polyalthia Nemoralis Extract Reverses the Chemoresistance to Cisplatin in Human Ovary Cancer Cells

目的:探讨陵水暗罗提取物暗罗素(ZPT)对耐药卵巢癌细胞株顺铂(DDP)敏感性的影响。方法:MTS法检测暗罗素和顺铂对卵巢癌细胞株A2780和耐药株A2780/DDP的增殖抑制效应。Western Blot法检测A2780和A2780/DDP细胞株p65蛋白表达水平,荧光素酶法检测A2780和A2780/DDP细胞株NF-κB活性。结果:暗罗素可以显著抑制人卵巢癌细胞株A2780和耐药株A2780/DDP的增殖活性[A2780和A2780/DDP的半数抑制浓度(IC50)值分别为1.498μmol/L和1.516μmol/L];联合组(0.5μmol/L暗罗素+5μmol/L顺铂)相对于顺铂单药组,显著下调A2780/DDP的增殖活性(80.80%vs 35.63%,P<0.01)。荧光素酶法检测A2780和A2780/DDP细胞株的NF-κB活性显示,A2780/DDP组NF-κB活性是A2780组的1.92倍(P<0.01);Western Blot法检测p65蛋白表达水平,提示A2780/DDP组p65蛋白表达显著高于A2780组。相对于对照组,0.5μmol/L暗罗素可以显著下调NF-κB活性(100%vs 39.69%,P<0.01)。Western Blot法检查0.5μmol/L暗罗素对A2780/DDP抗凋亡蛋白Bcl-2的影响,发现暗罗素可以显著抑制Bcl-2蛋白表达。结论:陵水暗罗提取物暗罗素可以增加顺铂耐药细胞株A2780/DDP对顺铂的敏感性,暗罗素下调NF-κB-Bcl-2途径活性可能是其逆转顺铂耐药机制。

Objective:To investigate the effect o Zincpolyanemine extacted from Chinese herbs Polyalthia Nemoralis on the sensitivity of cisplatin in human ovary cancer cells.Methods:MTS was introduced to detect the cell viability of A278 and A2780/DDP cell lines;luciferase reporter system was applied to measure the NF-κB transcript activity and Western Blot was used to test the protein expression level of p65 and Bcl-2.Results:Zincpolyanemine(ZPT)inhibited the proliferation activity of A278 and A2780/DDP cell lines(the IC50 for A278and A2780/DDP:1.498μmol/L and 1.516μmol/L,respectively).Compared with Cisplatin group,combination of 0.5μmol/L ZPT and 5μmol/L Cisplatin group significantly enhanced the antitumor activity of Cisplatin in chemoresistant ovary cancer cell line A2780/DDP(80.80%vs 35.63%,P<0.01).NF-κB transcript activityin A2780/DDP cell line was 1.92 fold of that in A2780 cell line(P<0.01)detected by luciferase reporter system.Western Blot assay showed that 0.5μmol/L ZPT downregulated the protein expression level of p65 and Bcl-2.Conclusion:Zincpolyanemine extracted from Chinese herbs Polyalthia Nemoralis may reverse the chemoresistance to cisplatin in human ovary cancer cells,which might be mediated via the inhibition of NF-κB-Bcl-2pathway.