白花丹醌对大细胞肺癌NL9980细胞的抗肿瘤作用及机制  被引量:4

Research of the inhibitory effect and the mechanisms of plumbagin on large cell lung cancer NL9980 cell line

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作  者:于涛[1] 李凯钰 宫婷[1] 邵宜[1] 刘刚[2] 

机构地区:[1]天津医科大学总医院肿瘤内科,300052 [2]天津医科大学总医院普通外科,300052

出  处:《中华实验外科杂志》2018年第2期271-274,共4页Chinese Journal of Experimental Surgery

摘  要:目的观察白花丹醌对大细胞肺癌NL9980细胞株的抗肿瘤作用,探讨白细胞介素-6/信号转导和转录激活因子3(IL-6/STAT3)信号通路在其抗肿瘤过程中的作用及机制。方法采用不同浓度白花丹醌处理NL9980细胞株,噻唑蓝(MTT)法观察其对肿瘤细胞的抑制效果。采用简单随机抽样法设置白花丹醌不同浓度组(5.0、7.5、10.0 μmol/L)及空白对照组(不予白花丹醌处理),培养24 h后收集细胞。实时定量聚合酶链反应(Real-time PCR)检测IL-6和STAT3的mRNA表达。酶联免疫吸附试验(ELISA)法、Western blot分别检测培养细胞上清液内IL-6蛋白表达和细胞内磷酸化STAT3(p-STAT3)蛋白表达。Real-time PCR检测通路下游调控基因B细胞淋巴瘤/白血病-2(bcl-2)、血管内皮生长因子(VEGF)、细胞周期蛋白D1(Cyclin D1)的mRNA水平。结果MTT法结果显示白花丹醌对大细胞肺癌NL9980细胞具有明显抑制作用,随药物浓度升高,抑制作用更为显著(F=32.17,P=0.000)。与对照组比较,白花丹醌干预后各组IL-6和STAT3的mRNA及其蛋白表达量均减少。随着白花丹醌浓度增加,IL-6 mRNA(分别为0.50±0.02、0.44±0.1、0.34±0.04,F=173.067,P=0.000)及蛋白(分别为4.71±0.02、4.08±0.14、2.01±0.07,F=1 805.177,P=0.000)表达量降低;STAT3 mRNA(分别为0.86±0.08、0.72±0.03、0.52±0.06,F=230.918,P=0.000)及蛋白(分别为0.65±0.19、0.45±0.22、0.31±0.11,F=17.332,P=0.000)表达量随着白花丹醌浓度增加表达量明显降低,组间差异均有统计学意义。与对照组比较,白花丹醌处理组中通路下游基因bcl-2、VEGF、Cyclin D1的mRNA表达量均降低,且随药物浓度增加降低更加明显,差异有统计学意义(F=33.698,P=0.000;F=337.743,P=0.000;F=180.136,P=0.000)。相关性分析结果表明IL-6和SATA3与bcl-2、VEGF、Cyclin D1均呈正相关。结论白花丹Objective To investigate the effects of plumbagin on human NL9980 large-cell lung cancer cells and the role of interleukin-6/signal transducers and activators of transcription 3 (IL-6/STAT3) signaling pathway in the anticancer procedure of plumbagin.Methods Different concentrations of plumbagin were administrated in NL9980 cell line. Inhibitory effects of plumbagin on NL9980 cells were analyzed using methyl thiazol tetrazolium (MTT) assay. Groups of different drug doses (5.0, 7.5, 10.0 μmol/L) and blank control (without administration of plumbagin) were set with simple random sampling method. Cells were collected after 24 h of administration. Expression of IL-6 and STAT3 and the downstream genes B-cell lymphoma/leukemia-2 (bcl-2), vascular endothelial growth factor (VEGF), and Cyclin D1 were assessed by real-time quantitative polymerase chain reaction (Real-time PCR). Enzyme linked immunosorbent assay (ELISA) was used to measure secreted IL-6 levels and western blotting to measure intracellular phospho-STAT3 levels.Results Plumbagin significantly inhibited NL9980 cells via suppressing IL-6/STAT3 signaling. Expression of all genes was inhibited by plumbagin in a dose-dependent manner (F=32.170, P=0.000). As the concentration of plumbagin increased, the expression of IL-6 mRNA decreased significantly (F=173.067, P=0.000) to 0.50±0.02, 0.44±0.1, 0.34±0.04 respectively and IL-6 protein decreased to 4.71±0.02, 4.08±0.14, 2.01±0.07 significantlly (F=1 805.177, P=0.000). Besides, the expression of STAT3 mRNA (0.86±0.08, 0.72±0.03, 0.52±0.06, F=230.918, P=0.000) and protein (0.65±0.19, 0.45±0.22, 0.31±0.11, F=17.332, P=0.000) were reduced with the increase of concentration of plumbagin. Compared with the control group, expression of downstream genes bcl-2, VEGF and Cyclin D1 mRNA were decreased significantly after administration of plumbagin (F=33.698, P=0.000; F=337.743, P=0.000; F=180.136, P=0.000), and declined gradually as drug concentration increas

关 键 词:白花丹醌 白细胞介素-6 信号转导和转录激活因子3 大细胞肺癌 

分 类 号:R285[医药卫生—中药学]

 

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