X线照射对U87胶质瘤多细胞球体MMP-2活性及Cho/Cr的影响

doi:10.6040/j.issn.1671-7554.0.2015.1000

摘要/Abstract

摘要： 目的构建胶质瘤细胞系U87的多细胞球体(MCS)模型,探讨不同剂量的X线对胶质瘤多细胞球体上清液基质金属蛋白酶2(MMP-2)活性和水溶性代谢物的影响。方法采用琼脂糖培养法建立U87多细胞球体模型;用0、1、5、10 Gy的X线分别照射多细胞球体,0 Gy组作为对照组,利用高氯酸提取细胞中水溶性代谢物［胆碱(Cho)、肌酸(Cr)、N-乙酰天门冬氨酸(NAA)］,采用14.7T高场强磁共振仪获取水溶性代谢物波谱;采用明胶酶谱法测量U87多细胞球体上清中MMP-2活性。结果成功建立了U87多细胞球体模型;多细胞球体在1、5、10 Gy X线照射下的胆碱与肌酸比值(Cho/Cr)分别为2.74±0.02、4.32±0.17、4.92±0.07,与对照组(2.10±0.29)比较,细胞内水溶性代谢物比值Cho/Cr升高(P=0.033, P=0.000, P=0.000);多细胞球体在0~10 Gy X线照射范围内,随着照射剂量的增加,细胞MMP-2活性增加。与对照组比较,差异有统计学意义(P=0.000)。结论亚致死剂量的X线可导致U87胶质瘤多细胞球体MMP-2活性增高,Cho/Cr升高。

关键词: 基质金属蛋白酶2, 氢质子磁共振波谱, 胆碱/肌酸, 胶质瘤, 多细胞球体

Abstract: Objective To establish the U87 glioma multicellular spheroid(MCS)model and to explore the effect of various dosages of X-ray on the activity of MMP-2 and water-soluble metabolites of U87 glioma multicellular spheroids. Methods The U87 glioma MCS model was established and irradiated by X-ray of 0, 1, 5 and 10 Gy. The 0 Gg group served as the control group. Perchloric acid extracting(PCA)method was used to extract water-soluble metabolites, including choline(Cho), creatine(Cr), and N-acetylaspartate(NAA), which were observed using in vitro 14.7T high resolution ¹H-MRS. The MMP-2 expression in U87 MCS model was detected with gelatin zymography. Results U87 glioma MCS model was established successfully. With the increase of X-ray doses, the Cho/Cr ratio rose from 2.74±0.02(1 Gy), 4.32±0.17(5 Gy)to 4.92±0.07(10 Gy); compared with the control group(2.10±0.29), the Cho/Cr ratio increased in the other groups［P=0.033(1 Gy), P=0.000(5 Gy), P=0.000(10 Gy)］. Compared with the control group, the activity of MMP-2 also increased(P=0.000). Conclusion Sublethal irradiation promotes the activity of MMP-2 of U87 MCS and elevates the Cho/Cr ratio.

Key words: Glioma, Multicellular spheroid, Proton magnetic resonance spectroscopy, Matrix metalloproteinase 2, Cho/Cr

中图分类号:

R445.2

徐延杰,崔谊,李红霞,史文琦,李福艳,王建震,曾庆师. X线照射对U87胶质瘤多细胞球体MMP-2活性及Cho/Cr的影响[J]. 山东大学学报（医学版）, 2016, 54(4): 6-10.

XU Yanjie, CUI Yi, LI Hongxia, SHI Wenqi, LI Fuyan, WANG Jianzhen, ZENG Qingshi. Effect of X-ray on the activity of MMP-2 and Cho/Cr ratio of U87 glioma multicellular spheroid[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2016, 54(4): 6-10.

参考文献

[1] Chinot OL, Wick W, Mason W, et al. Bevacizumab plus radiotherapy-temozolomide for newly diagnosed glioblastoma[J]. N Engl J Med, 2014, 370(8): 709-722.
[2] Badiga AV, Chetty C, Kesanakurti D, et al. Mmp-2 sirna inhibits radiation-enhanced invasiveness in glioma cells[J]. PloS One, 2011, 6(6): e20614. doi: 10.1371/journal.pone.0020614.
[3] Pei J, Park IH, Ryu HH, et al. Sublethal dose of irradiation enhances invasion of malignant glioma cells through p53-mmp 2 pathway in U87MG mouse brain tumor model[J]. Radiat Oncol, 2015, 10: 11. doi: 10.1186/s13014-015-0475-8.
[4] 高峰. 制备和培养多细胞肿瘤球体的简易方法[J]. 黑龙江医学, 2001, 25(7): 481-482. GAO Feng. Simple method for preparation and culture of multi -cellular tumor spheroid[J]. Hei Long Jiang Medical Journal, 2001, 25(7): 481-482.
[5] Xu ZF, Pan AZ, Yong F, et al. Human umbilical mesenchymal stem cell and its adipogenic differentiation: Profiling by nuclear magnetic resonance spectroscopy[J]. World J Stem Cells, 2012, 4(4): 21-27.
[6] Cao Z, Wu LP, Li YX, et al. Change of choline compounds in sodium selenite-induced apoptosis of rats used as quantitative analysis by in vitro 9.4t mr spectroscopy[J]. World J Gastroenterol, 2008, 14(24): 3891-3896.
[7] Jagtap JC, Parveen D, Shah RD, et al. Secretory prostate apoptosis response(par)-4 sensitizes multicellular spheroids(mcs)of glioblastoma multiforme cells to tamoxifen-induced cell death[J]. FEBS Open Bio, 2015, 5: 8-19. doi: 10.1016/j.fob.2014.11.005.
[8] Carlsson J. Proliferation gradient in three dimensional colonies of cultured human glioma cells[J]. Int J Cancer, 1977, 20(1): 129-136.
[9] Freyer JP, Sutheriand RM. Proliferative and clonogeneity of cells from EMT6/RO multicellular spheroids induced by the glucose and oxygen supply[J]. Cancer Res, 1986, 46(7): 3513-3520.
[10] Sainan C, Qiang L, Yubo L, et al. Correlation of fractional anisotropy and metabolite concentrations measured using 1h-mrs of cerebral white matter in healthy adults[J]. Biomed Mater Eng, 2014, 24: 3017-3024.
[11] Zhang Z, Zeng Q, Liu Y, et al. Assessment of the intrinsic radiosensitivity of glioma cells and monitoring of metabolite ratio changes after irradiation by 14.7-T high-resolution 1H MRS[J]. NMR Biomed, 2014, 27(5): 547-552.
[12] Doblas S, He T, Saunders D, et al. In vivo characterization of several rodent glioma models by 1H MRS[J]. NMR Biomed, 2012, 25(4): 685-694.
[13] Guan L, Li ZY, Zhang YL, et al. Early biomarkers in 1h nuclear magnetic resonance spectroscopy of striatal pathological mechanisms after acute carbon monoxide poisoning in rats[J]. Biomed Environ Sci, 2015, 28(10): 728-737.
[14] Kinoshita Y, Yokota A. Absolute concentrations of metabolites in human brain tumors using in vitro proton magnetic resonance spectroscopy[J]. NMR Biomed, 1997, 10(1): 2-12.
[15] Jarskog LF, Dong Z, Kangarlu A, et al. Effects of davunetide on N-acetylaspartate and choline in dorsolateral prefrontal cortex in patients with schizophrenia[J]. Neuropsychopharmacology, 2013, 38(7): 1245-1252.
[16] Tarnawski R, Sokol M, Pieniazek P, et al. H-1-mrs in vivo predicts the early treatment outcome of postoperative radiotherapy for malignant gliomas[J]. Int J Radiat Oncol Biol Phys, 2002, 52(5): 1271-1276.
[17] 刘伟, 王长坤, 只达石. MMP-2、MMP-9在人脑胶质瘤及其浸润组织中的表达和临床意义[J].中风与神经疾病杂志, 2009, 26(6): 689-692. LIU Wei, WANG Changkun, ZHI Dashi. The expression and clinical significance of the MMP-2 and MMP-9 in human glioma and gliomas invasive tissue[J]. J Apoplexy and Nervous Diseases, 2009, 26(6): 689-692.
[18] Wild-Bode C, Weller M, Rimner A, et al. Sublethal irradiation promotes migration and invasiveness of glioma cells: implications for radiotherapy of human glioblastoma[J]. Cancer Res, 2001, 61(6): 2744-2750.

相关文章 15

[1]	王琳琳孙美丽孙玉萍张楠刘传勇. 中心体α-微管蛋白、γ-微管蛋白在脑胶质瘤中的表达及其与Survivin表达的相关性研究[J]. 山东大学学报(医学版), 2209, 47(6): 103-.
[2]	李刚,薛皓,邱伟,赵荣荣. 脑胶质瘤抑制性免疫微环境形成机制及研究进展[J]. 山东大学学报 (医学版), 2020, 1(8): 67-73.
[3]	吴强,何泽鲲,刘琚,崔晓萌,孙双,石伟. 基于机器学习的脑胶质瘤多模态影像分析[J]. 山东大学学报 (医学版), 2020, 1(8): 81-87.
[4]	陈安静,张训. 靶向小类泛素化修饰的胶质瘤治疗新策略[J]. 山东大学学报 (医学版), 2020, 1(8): 88-94.
[5]	江涛. 类脑智能在脑科学的前沿应用[J]. 山东大学学报 (医学版), 2020, 1(8): 10-13.
[6]	徐继禧,陈伟健. 髓内弥漫性中线胶质瘤伴H3 K27M突变1例[J]. 山东大学学报 (医学版), 2020, 1(7): 96-101.
[7]	王超超,田海龙,姜慧峰,郭文强,王志刚. 荧光素钠“黄荧光”导航辅助高级别胶质瘤手术治疗的临床意义[J]. 山东大学学报（医学版）, 2017, 55(11): 32-37.
[8]	巨媛媛,任满意,李睿,赵萌萌,隋树建. TWEAK通过ERK1/2通路促进大鼠心肌成纤维细胞MMP2与Ⅰ型胶原表达[J]. 山东大学学报（医学版）, 2016, 54(5): 23-28.
[9]	葛晓燕,刘颖,李怡,王翠兰. 富亮氨酸胶质瘤失活1蛋白抗体阳性相关的边缘性脑炎1例[J]. 山东大学学报（医学版）, 2016, 54(10): 92-94.
[10]	姚众, 陈为亮, 徐洋洋, 何影, 曲迅, 李新钢. 半乳凝素-9抑制胶质瘤细胞增殖及迁移[J]. 山东大学学报（医学版）, 2015, 53(4): 1-5.
[11]	朱晓东, 夏同良, 姚庆宇, 李昊元, 王本琳, 倪石磊, 王建刚, 李新钢, 苏万东. SN-38缓释系统的制作及其抗胶质瘤作用的体外实验研究[J]. 山东大学学报（医学版）, 2015, 53(4): 6-11.
[12]	邢德广, 马二猛, 王谋龙, 孙中正, 樊明德, 臧贻征, 王成伟. 紫杉醇协同肿瘤坏死因子相关凋亡诱导配体抑制胶质瘤细胞活性的探讨[J]. 山东大学学报（医学版）, 2015, 53(4): 12-15.
[13]	杨慧, 李万湖, 陈月芹, 郭沐洁, 徐亮, 吴玉芬. 3.0T¹H-MRS鉴别高级别胶质瘤与单发性脑转移瘤的价值[J]. 山东大学学报（医学版）, 2015, 53(4): 65-70.
[14]	胡芳志, 张正军, 耿厚法, 梁秋华, 孙琳. 2型糖尿病患者尿微量白蛋白与脑组织代谢物变化的关系[J]. 山东大学学报（医学版）, 2015, 53(2): 43-47.
[15]	张照涛1,张华2,何敬振1,朱建辉3,刘芸1,王建震1,李传福1,曾庆师1. 高场强磁共振波谱观察脑胶质瘤细胞代谢特征[J]. 山东大学学报（医学版）, 2014, 52(3): 19-22.

多维度评价

Viewed

Full text

Abstract

Cited

Shared

Discussed