槐定碱对神经胶质瘤U87细胞增殖、侵袭及相关信号通路的影响  

Effect of sophoridine on proliferation,invasion and relative signaling pathways of neuroglioma U87 cell

作　　者：赵树鹏[1] 靳彩玲[2] 高国军[1] 赵新利[1] 金保哲[1] 周文科[1] 

机构地区：[1]新乡医学院第一附属医院神经外科,河南卫辉453100 [2]新乡医学院第一附属医院肿瘤科,河南卫辉453100

出　　处：《中国肿瘤生物治疗杂志》2016年第3期360-365,共6页Chinese Journal of Cancer Biotherapy

基　　金：河南省医学科技攻关计划基金资助项目(No.2011020091)~~

摘　　要：目的:探讨槐定碱对神经胶质瘤U87细胞增殖和侵袭的抑制作用以及U87细胞DNA拓扑异构酶I(DNA TOP I)、EGFR-酪氨酸激酶(EGFR-TK)、基质金属蛋白酶2(MMP2)和氨肽酶N(APN)活性的影响。方法:将不同浓度槐定碱加入到神经胶质瘤U87细胞株中,利用MTT法测定槐定碱对U87细胞和人脑正常星型胶质HEB细胞生长的抑制作用,酶标仪法检测槐定碱对细胞凋亡蛋白caspase-3活性,采用Transwell小室法分析U87细胞的侵袭能力;采用非放射性NF-κB EMSA试剂盒测定U87细胞中NF-κB表达。结果:随着槐定碱浓度的增加(5、10、25、50、100μmol/L),神经胶质瘤U87细胞生长抑制率不断的增加,但HEB细胞的生长并未受到明显的抑制[(11.23±1.18)%vs(2.43±0.29)%、(22.48±3.21)%vs(3.65±0.42)%、(43.21±4.09)%vs(4.03±0.55)%、(57.31±5.09)%vs(5.21±0.43)%、(77.98±6.98)%vs(7.22±0.78)%,均P<0.05];与空白组比较,槐定碱存在下的U87细胞侵袭能力明显降低[(87.43±7.33)%、(65.12±6.16)%、(50.63±4.56)%、(35.32±4.04)%、(23.46±2.32)%vs(120.32±9.32),均P<0.05]。槐定碱对DNA TOP I、EGFR-TK、APN和MMP-2的半抑制率IC_(50)分别为(22.43±2.21)、(31.25±3.09)、(6.32±0.32)和(8.23±0.63)μmol/L,但U87细胞中凋亡蛋白caspase-3活性呈增强趋势;槐定碱能够下调NF-κB信号的表达。结论:低毒性的槐定碱可能通过降低DNA TOP I、EGFR-TK、APN和MMP-2活性,并下调NF-κB信号通路和激活凋亡caspase-3酶联反应的方式,对神经胶质瘤U87细胞的侵袭、增殖和信号通路产生抑制作用。Objective： To explore inhibitory effects of sophoridine on DNA topoisomerase I（ DNA TOP I）,epidermal growth factor receptor-tyrosine kinase（ EGFR-TK）,matrix metalloproteinase 2（ MMP-2） and aminopeptidase N（ APN）,and the machanism for that growth and invasion of neuroglioma U87 cells were inhibited by the sophoridine. Methods： Sophoridine at different concentrations were added into brain neuroglioma U87 line cells. Inhibitory effect of the sophoridine on growths of the U87 cells and HEB human brain astrocytes was determined with MTT assay,activity of the sophoridine on apoptosis-associated protein caspase-3 in the U87 cells was detected by enzyme-linked turbidimetry assay,Transwell tests were used to determe effect of the sophoridine on invasion ability of the U87 cells. Expression of NF-κB in the U87 cells was examed by a non radioactive EMSA reagent kit. Results： Growth inhibitory rate of the neuroglioma U87 cells increased constantly with increase of sophoridine concentration（ 5,10,25,50 and 100μmol / L）,but growth rate of the HEB cells did not inhibited obviously（ [11. 23 ± 1. 18]% vs [2. 43 ± 0. 29]%,[22. 48 ± 3. 21]% vs [3. 65 ±0. 42]%,[43. 21 ± 4. 09]% vs [4. 03 ± 0. 55]%,[57. 31 ± 5. 09]% vs [5. 21 ± 0. 43]%,[77. 98 ± 6. 9]% vs[7. 22 ± 0. 78]%,all P〈0. 05）. Compared with the blank control group,invasion ability of the U87 cells was significantly inhibited as presence of the sorphoridine（ [87. 43 ± 7. 33]%,[65. 12 ± 6. 16]%,[50. 63 ± 4. 56]%,[35. 32 ± 4. 04]%,[23. 46 ± 2. 32]% vs [120. 32 ± 9. 32]%,all P〈0. 05）. Half inhibitory rates（ IC_（50）） of the sorphoridin on DNA TOP I,EGFR-TPK,APN and MMP-2 were（ [22. 43 ± 2. 21],[31. 25 ± 3. 09],[6. 32 ± 0. 32] and[8. 23 ± 0. 63]μmol /L） respectively. But activity of apoptosis protein caspase-3 in the U87 cell was enhanced and the sorphoridin down-regulated expression of NF-κB signaling. Conclution： The sorphoridin with low toxicity could reduce activities of DNA TOP I,EGFR-TPK,APN and MMP-2

关 键 词：槐定碱 神经胶质瘤U87细胞 DNA拓扑异构酶I 表皮生长因子受体-酪氨酸激酶 基质金属蛋白酶2 氨肽酶N 

分 类 号：R739.41[医药卫生—肿瘤] R730.5[医药卫生—临床医学]