目的 探讨姜黄素对人脑胶质瘤U251细胞侵袭与迁移的抑制作用。方法 采用四甲基偶氮唑蓝法筛选姜黄素实验浓度;采用细胞划痕实验观察姜黄素作用0、6、12和24 h后胶质瘤U251细胞的迁移情况,并且测量24 h后迁移的距离;采用Transwell实验观察穿膜细胞数,以判断细胞的侵袭能力;采用Western blotting实验检测胶质瘤U251细胞中侵袭相关蛋白基质金属蛋白酶2(matrix metalloproteinase-2, MMP2)和基质金属蛋白酶9的表达。结果 四甲基偶氮唑蓝法筛选的姜黄素浓度为10和20 μmolL-1;与空白对照组相比,在姜黄素作用24 h后,胶质瘤U251细胞迁移的距离和穿膜细胞数皆明显减少,且与药物浓度呈正相关(P＜0.01);与空白对照组相比,姜黄素组胶质瘤U251细胞内基质金属蛋白酶2和基质金属蛋白酶9蛋白的表达明显下降(P＜0.01),在姜黄素10和20 μmolL-1组基质金属蛋白酶2和基质金属蛋白酶9蛋白的表达差异明显(P＜0.05)。结论 姜黄素能够抑制人脑胶质瘤U251细胞的迁移与侵袭,可能是通过下调细胞内基质金属蛋白酶2和基质金属蛋白酶9蛋白的表达引起的。
Abstract
OBJECTIVE To investigate the inhibitory effect of curcumin on invasion and migration of human glioma U251 cells. METHODS Curcumin experimental concentration was screened by MTT. Through cell scratch assay, migration distance of glioma U251 cells was observed after using curcumin for 0, 6,12 and 24 h, migration distance of cells was measured after treatment with curcumin for 24 h . Through transwell assay, transmembrane cell number was recorded and to judge invasive ability of cells. Through Western blotting assay, invasion related proteins matrix metalloproteinase 2 (MMP2) and MMP9 were detected in the glioma U251 cells. RESULTS Screening curcumin concentration was 10 and 20 μmolL-1 by MTT. Compared with the control group, the migration distance and transmembrane cell number of glioma U251 cells decreased significantly after culturing cells for 24 h with curcumin, which were positively correlated with the concentration of drug(P＜0.01). Compared with the control group,the expression of MMP2 and MMP9 proteins of glioma U251 cells decreased significantly in curcumin groups(P＜0.01). Compared with the curcumin 10 μmolL-1 group,the expression of MMP2 and MMP9 proteins decreased significantly in the curcumin 20 μmolL-1 group(P＜0.05). CONCLUSION Curcumin can inhibit the migration and invasion of human glioma U251 cells, and it is possible that the result is relevant to the down-regulation of the expression of MMP2 and MMP9 proteins.
关键词
胶质瘤 /
姜黄素 /
细胞迁移 /
细胞侵袭 /
基质金属蛋白酶
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Key words
glioma /
curcumin /
cell migration /
cell invasion /
matrix metalloproteinase
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中图分类号：
R739.4
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参考文献
[1] FISHER J L, SCHWARTZBAUM J A, WRENSCH M, et al. Epidemiology of brain tumors[J]. Neurol Clin, 2007,25(4):867-890.[2] JEMAL A, SIEGEL R, WARD E, et al. Cancer statistics, 2007[J]. CA Cancer J Clin, 2007,57(1):43-66.[3] ZALUTSKY M R. Current status of therapy of solid tumors: Brain tumor therapy[J]. J Nucl Med, 2005,46(suppl 1):151-156.[4] SANAI N, BERGER M S. Operative techniques for gliomas and the value of extent of resection[J]. Neurotherapeutics, 2009,6(3):478-486. [5] PANG B C, WAN W H, LEE C K, et al. The role of surgery in high-grade glioma--is surgical resection justified A review of the current knowledge[J].Ann Acad Med Singapore, 2007,36(5):358-363.[6] SHUMAN MOSS L A, JENSEN-TAUBMAN S, STETLER-STEVENSON W G.Matrix metalloproteinases:Changing roles in tumor progression and metastasis[J].Am J Pathol, 2012,181(6):1895-1899.[7] KESSENBROCK K, PLAKS V, WERB Z. Matrix metalloproteinases: Regulators of the tumor microenvironment[J]. Cell, 2010,141(1):52-67.[8] KUTTAN R, BHANUMATHY P, NIRMALA K, et al. Potential anticancer activity of turmeric(Curcuma longa)[J]. Cancer Lett, 1985,29(2):197-202.[9] AGGARWAL B B, HARIKUMAR K B. Potential therapeutic effects of curcumin, the anti-inflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases[J]. Int J Biochem Cell Biol, 2009,41(1):40-59.[10] LIN S S, HUANG H P, YANG J S, et al. DNA damage and endoplasmic reticulum stress mediated curcumin-induced cell cycle arrest and apoptosis in human lung carcinoma A-549 cells through the activation caspases cascade- and mitochondrial-dependent pathway[J]. Cancer Lett, 2008,272(1):77-90.[11] LIN S S, LAI K C, HSU S C, et al. Curcumin inhibits the migration and invasion of human A549 lung cancer cells through the inhibition of matrix metalloproteinase-2 and -9 and Vascular Endothelial Growth Factor (VEGF)[J]. Cancer Lett, 2009,285(2):127-133.[12] DUVOIX A, BLASIUS R, DELHALLE S, et al. Chemopreventive and therapeutic effects of curcumin[J]. Cancer Lett, 2005,223(2):181-190.[13] WILKEN R, VEENA M S, WANG M B, et al. Curcumin: A review of anti-cancer properties and therapeutic activity in head and neck squamous cell carcinoma[J].Mol Cancer, 2011,10:12.[14] PERRY M C, DEMEULE M, RGINA A, et al. Curcumin inhibits tumor growth and angiogenesis in glioblastoma xenografts[J]. Mol Nutr Food Res, 2010,54(8):1192-1201.[15] WU Z M, YUAN X H, JIANG P C, et al. Curcum in induces U251 cell apoptosis via alteration of expression of FAK and activation of caspase [J]. Chin Pharmacol Bull(中国药理学通报),2006,22(4):484-487.[16] BALDUCCI M, D′AGOSTINO G R, MANFRIDA S, et al. Radiotherapy and concomitant temozolomide during the first and last weeks in high grade gliomas: Long-term analysis of a phase II study[J]. J Neurooncol, 2010,97(1):95-100.[17] GILBERT M R, GONZALEZ J, HUNTER K, et al. A phase I factorial design study of dose-dense temozolomide alone and in combination with thalidomide, isotretinoin, and/or celecoxib as postchemoradiation adjuvant therapy for newly diagnosed glioblastoma[J]. Neuro Oncol, 2010,12(11):1167-1172.[18] SHUMAN MOSS L A, JENSEN-TAUBMAN S, STETLER-STEVENSON W G . Matrix metalloproteinases: Changing roles in tumor progression and metastasis[J].Am J Pathol, 2012,181(16):1895-1899.[19] WATANABE H. Extracellular matrix-regulation of cancer invasion and metastasis[J]. Gan To Kagaku Ryoho, 2010,37(11):2058-2061.[20] RAVINDRAN J, PRASAD S, AGGARWAL B B. Curcumin and cancer cells: How many ways can curry kill tumor cells selectively[J]. AAPS J, 2009,11(3):495-510.[21] SHAO Z M, SHEN Z Z, LIU C H, et al. Curcumin exerts multiple suppressive effects on human breast carcinoma cells[J]. Int J Cancer, 2002,98(2):234-240.[22] SU C C, CHEN G W, LIN J G, et al. Curcumin inhibits cell migration of human colon cancer colo 205 cells through the inhibition of nuclear factor kappa B/p65 and down-regulates cyclooxygenase-2 and matrix metalloproteinase-2 expressions[J]. Anticancer Res, 2006,26(2A):1281-1288.
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脚注
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基金
国家自然科学基金资助项目(81270142)
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