β-榄香烯通过激活细胞内氧化还原系统、降低线粒体膜电位和 P 糖蛋白表达，诱导细胞凋亡，逆转 A549/DDP 肺癌耐药细胞的耐药性。
β-elemene reverses the drug resistance of A549/DDP lung cancer cells by activating intracellular redox system, decreasing mitochondrial membrane potential and P-glycoprotein expression, and inducing apoptosis.
机构信息
Department of Thoracic Surgery, the First Affiliated Hospital of Fujian Medical University Fuzhou, China; Department of Thoracic Surgery, Xiamen Traditional Chinese Medicine (TCM) Hospital of Fujian University of TCM Xiamen, China.
Department of Thoracic Surgery, the First Affiliated Hospital of Xiamen University Xiamen, China.
出版信息
Thorac Cancer. 2014 Jul;5(4):304-12. doi: 10.1111/1759-7714.12093. Epub 2014 Jul 3.
BACKGROUND
β-elemene (β-ELE) injection is a new anticancer drug extracted from Curcuma zedoaria Roscoe that has been widely used to treat malignant tumors. Recent studies show that β-ELE reverses the drug resistance of tumor cells. To explore the possible mechanisms of β-ELE, we investigated its effects on cisplatin (DDP)-resistant human lung adenocarcinoma A549/DDP cells.
METHODS
The effects of β-ELE on the growth of A549/DDP cells in vitro were determined by MTT assay. Apoptosis was assessed by fluorescence microscopy with Hoechst 33258 staining, flow cytometry with Annexin V-FITC/propium iodide double staining; mitochondrial membrane potential using JC-1 fluorescence probe and laser confocal scanning microscopy, and intracellular reactive oxygen species levels were measured by 2',7'-dichlorfluorescein-diacetate staining and flow cytometry; and contents of cytosolic glutathione were determined by glutathione assay kits. Intracellular Rhodamine-123 fluorescence intensity was detected by flow cytometry, and the expression of P-glycoprotein (P-gp) was detected by Western blotting.
RESULTS
β-ELE inhibited the proliferation of A549/DDP cells in a time- and dose-dependent manner. Furthermore, β-ELE enhanced the sensitivity of A549/DDP cells to cisplatin and reversed the drug resistance of A549/DDP cells. Consistent with a role in activating apoptosis, β-ELE decreased mitochondrial membrane potential, increased intracellular reactive oxygen species concentration and intracellular accumulation of Rhodamine-123, decreased the cytoplasmic glutathione levels and the expression of P-gp in a time- and dose-dependent manner.
CONCLUSIONS
These results define a pathway of β-ELE function that involves decreased mitochondrial membrane potential and P-gp expression activated intracellular redox system, and induced apoptosis leading to reverse drug resistance.
背景
β-榄香烯（β-ELE）注射液是一种从莪术中提取的新型抗癌药物，已广泛用于治疗恶性肿瘤。最近的研究表明，β-ELE 逆转了肿瘤细胞的耐药性。为了探讨β-ELE 的可能机制，我们研究了其对顺铂（DDP）耐药人肺腺癌细胞 A549/DDP 的影响。
方法
用 MTT 法测定β-ELE 对 A549/DDP 细胞体外生长的影响。用 Hoechst 33258 染色荧光显微镜观察细胞凋亡，Annexin V-FITC/碘化丙啶双染流式细胞术检测细胞凋亡；用 JC-1 荧光探针和激光共聚焦扫描显微镜检测线粒体膜电位；用 2',7'-二氯荧光素二乙酸酯染色和流式细胞术检测细胞内活性氧水平；用谷胱甘肽检测试剂盒检测细胞内谷胱甘肽含量。用流式细胞术检测细胞内罗丹明 123 荧光强度，用 Western blot 检测 P-糖蛋白（P-gp）的表达。
结果
β-ELE 呈时间和剂量依赖性抑制 A549/DDP 细胞的增殖。此外，β-ELE 增强了 A549/DDP 细胞对顺铂的敏感性，并逆转了 A549/DDP 细胞的耐药性。与激活细胞凋亡的作用一致，β-ELE 降低了线粒体膜电位，增加了细胞内活性氧浓度和罗丹明 123 的细胞内积累，降低了细胞质谷胱甘肽水平和 P-gp 的表达，呈时间和剂量依赖性。
结论
这些结果定义了β-ELE 功能的途径，涉及降低线粒体膜电位和 P-gp 表达激活细胞内氧化还原系统，并诱导细胞凋亡导致逆转耐药性。
Zotero 插件