姜黄素对人多发性骨髓瘤ARH-77细胞外源性凋亡通路的影响  

Effect of curcumin on extrinsic apoptosis pathway in human multiple myeloma cell ARH-77

作　　者：曲佳[1] 陈玲珍[1] 詹昱[1] 冯可欣[1] 杨郁青[1] 巫进明[1] 余卫[1] 

机构地区：[1]广州市第十二人民医院血液科,广东广州510282

出　　处：《中国肿瘤生物治疗杂志》2014年第1期25-30,共6页Chinese Journal of Cancer Biotherapy

基　　金：广州市医药卫生科技项目(No.20121A011102)~~

摘　　要：目的:探讨姜黄素(curcumin,Cur)对人多发性骨髓瘤ARH-77细胞外源性凋亡通路的影响。方法:ARH-77细胞经6.25、12.5、25、50、100、200μmol/L Cur处理12、24、48 h,MTT法检测Cur对ARH-77细胞增殖的抑制作用,Hoechst 33258染色法观察Cur处理24 h后ARH-77细胞凋亡的形态学改变,流式细胞术检测ARH-77细胞周期和Fas/FasL、TRAIL/TRAIL-R的表达,分光光度法检测ARH-77细胞caspase-8的活性。结果:Cur对ARH-77细胞的增殖有时间和剂量依赖的抑制作用。25μmol/L Cur处理ARH-77细胞可观察到凋亡小体,Cur阻滞细胞周期于G0/G1期,并且有凋亡峰。其促凋亡作用呈浓度依赖性,6.25、12.5、25μmol/L Cur作用24 h后,ARH-77细胞凋亡率均显著高于对照组[(10.35±0.35)%、(14.35±1.34)%、(36.65±1.06)%vs(3.83±0.32)%,F=500.432,P=0.000];实验组细胞内caspase 8的活化程度均显著高于对照组[(0.223±0.018)、(0.263±0.019)、(0.240±0.035)vs(0.154±0.007);F=9.059,P=20.03]。12.5μmol/L Cur作用24 h后,ARH-77细胞表面Fas[(99.05±0.49)%vs(92.10±0.70)%,t=15.404,P=0.000]、FasL[(9.05±0.78)%vs(1.73±1.19)%,t=9.487,P=0.008]、TRAIL[(1.35±0.07)%vs(0.55±0.07)%,t=-11.317,P=0.008]、DR4、DcR1和DcR2的表达均显著升高,DR5表达显著降低[(0.95±0.07)%vs(7.70±0.29)%,t=32.742,P=0.001];进一步提升Cur浓度至25μmol/L,却降低了DcR1[(4.35±1.20)%vs(14.25±0.21)%;t=5.692,P=0.008]及DcR2[(0.75±0.21)%vs(1.65±0.71)%;t=11.470,P=0.03]的表达。结论:Cur能明显抑制人多发性骨髓瘤ARH-77细胞的增殖,其机制可能与激活外源性凋亡通路从而诱导细胞凋亡有关。Objective：To investigate the influence of curcumin （Cur） on the extrinsic apoptosis pathway of human multiple myeloma cell line ARH-77. Methods： ARH-77 cells were treated with Cur at 6.25, 12.5, 25, 50, 100 and 200 μmol/L. At 12, 24 and 48 h after treatment, cell viability was analyzed by MTT assay and growth inhibition was accordingly calculated. At 24 h after treatment, changes in the cell morphology were assessed by Hoechst 33258 staining, cell cycle progression and levels of Fas/Fasl and TRAIL/TRAIL-R were analyzed by flow cytometry, and the activity of caspase 8 was determined by colorimetry. Results： Cur significantly inhibited the growth of ARH-77 cells in a time- and dose-dependent manner. At 24 h after treatment, Cur induced apoptosis in ARH-77 cells in a dose-dependent manner; the percentage of apoptotic cells was （10.35±0.35）% at 6.25 μmol/L, （1435±1.34）% at 12.5 μmol/L and （36.65±106）% at 25 μmol/L, significantly higher than that in untreated control cells （[3.83±0.32]%, P 〈0.01）. Apoptotic bodies and cell cycle arrest at the G0/G1 phase were seen in ARH-77cells treated with 25 μmol/L Cur. Caspase 8 activity was significantly higher in ARH-77 cells treated with Cur at 6.26 μmol/L（0.223±0.018）, 12.5 μmol/L （0.263±0.019）, or 25.0 μmol/L （0.240±0.035） than in untreated control cells （0.154±0.007） （ P 〈0.05）. Compared with the non-treatment control, 24 h Cur treatment at 6.25 μmol/L significantly increased the protein levels of Fas （[99.05±0.49]% vs [92.10±0.7]%, P ＝0.000）, FasL （[9.05±0.78]% vs [1.73±1.19]%, P ＝0008）, TRAIL （[1.35±0.07]% vs [0.55±0.07]%, P ＝0.008）, DR4, DcR1 and DcR2 but significantly decreased DR5 （[0.95±0.07]% vs [7.70±0.29]%, P ＝0.001）. The effect of Cur on DcR1 （[4.35±1.20]% vs [14.25±0.21]%, P ＝0.008） and DcR2 （[0.75±0.21]% vs [1.65±0.71]%, P ＝0.03） were more pronounced at 25.0 μmol/L than at 12.5 μmol/L. Conclusion： Cur is

关 键 词：姜黄素 多发性骨髓瘤 凋亡通路 FAS TRAIL 

分 类 号：R733.3[医药卫生—肿瘤]