摘要
目的探讨脑胶质瘤微环境及环氧化酶2抑制剂塞来昔布对树突状细胞（dendritic cell，DC）表型和功能的影响。方法检测塞来昔布对C6细胞COX-2蛋白表达及PGE2分泌水平的影响。分离人外周血单个核细胞，分为正常诱导组、c6细胞上清组及塞来昔布＋C6细胞上清组，体外诱导培养为DC细胞。观察DC形态学变化，检测DC表面分子CD80、CD83、CD86、CD1a的表达。检测DC培养上清中IL-12的水平及其体外刺激同种异体T细胞增殖能力。结果塞来昔布可呈浓度依赖性下调c6细胞COX-2蛋白表达水平及PGE2分泌水平。各组细胞均可诱导出典型DC形态。三组DCCD1a表达率（％）为（75．56±2．40，75．09±3．67，76．03±3．43），CD83表达率（％）为（72．04±3．45，71．44±3．78，73．63±3．31），差异无统计学意义（尸〉0．05），C6上清组DCCD80、CD86表达率（％）分别为（58．41-I-3．85，58．22±3．25），低于正常诱导组（分别为70．36±2．91，69．31±4．29），差异具有统计学意义（尸〈0．01），对应其刺激T淋巴细胞增殖能力降低（P〈0．01），IL-12分泌水平（pg／ml）低于正常诱导组（分别为137．88±5．33，186．04±4．76），差异具有统计学意义（P〈0．01）。塞来昔布预处理C6细胞后可提高CD80、CD86表达水平（66．83±2．51，63．51±5．47，P〈0．01）及刺激T淋巴细胞增殖能力、IL-12分泌水平（170．31±3．46，P〈0．01），但仍低于正常诱导组（P〈0．01）。结论塞来昔布可通过抑制COX-2蛋白活性，降低肿瘤局部微环境中的PGE2水平，改善DC的表型和功能。
Objective To investigate the effects of glioma microenvironment and COX-2 inhibitor celecoxib on the phenotypes and function of dendritic cell（DC）. Methods The expression of cyclooxygenase-2 （COX-2） and prostaglandin E2（PGE2） production were detected in glioma C6 cells treated with different concentration of celecoxib. Monocytes were isolated from human peripheral blood and cultured with 200 ng/ml rhGM-CSF and 50 ng/ml rhlL-4, either C6 tumor cells supernatant （TSN） or TSN from C6 cells treated with celecoxib to generate DCs. Cell morphology was observed. Cell phenotype including CDla, CD80, CD83 and D86 were analyzed on a FACScan. Production of IL-12 in DC supernatant and the potential to stmiulate allogeneic T cell proliferation were detected. Results The expression of COX-2 and PGE2 production in C6 cells decreased after treated with celecoxib in a concentration dependant manner. Typical DCs were induced in all groups and the expression of CD1 a （（75.56 ±2.40）% ,（75.09 ±3.67）% ,（76.03 ± 3.43）% ） ,CD83（ （72.04 ±3.45）%, （71.44 ± 3.78）%, （73.63 ±3.31 ） % ） had no difference （P 〉 0.05 ）. Expression of CD80 （ （58.41±3.85 ） % ）, CD86 （ （58.22 ± 3.25） % ） in DC with TSN obviously decreased compared with normal group （ 70.36 ±2.91 ）%, （69.31± 4.29 ） %, P 〈 0.01 ） as well as the IL-12 production （ 137.88 ± 5.33 ） pg/ml, （ 186.04 ±4.76 ） pg/ml ） and the potential to stmiulate allogeneic T cell proliferation （ P 〈 0. 01 ）. Celecoxib increased the expression of CD80, CD86 （ 66.83 ± 2.51,63.51 ） 5.47, P 〈 0.01 ） in DC and the same as IL-12 production （ 170.31 ± 3.46 ） pg/ml, P 〈0. 01 ） and the potential to stmiulate allogeneic T cell proliferation （ P 〈 0.01 ）, which were lower than the normal level. Conclusion Glioma microenvironment may induce the celecoxib can inhibit the expression of COX-2 and PGE2 in gliomas ceils and improve the phenotypes and function defect of DCs.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2012年第10期880-883,共4页
Chinese Journal of Behavioral Medicine and Brain Science
基金
山东省自然科学基金项目（ZR2009CQ015）
