颅咽管瘤组织内的炎性反应及其临床意义

Inflammatory response in craniopharyngiomas and its clinical significance

目的探究颅咽管瘤组织内的炎性反应及其临床意义。方法应用抗体芯片技术分析颅咽管瘤及鞍区其他常见肿瘤（包括下丘脑胶质瘤、垂体腺瘤、脑膜瘤）组织中炎性因子表达，通过实时荧光定量PCR（qPCR）、酶联免疫吸附实验（ELISA）进一步验证。利用颅咽管瘤原代细胞的培养上清处理小鼠脑神经元细胞株（CATH．a），流式细胞技术检测细胞凋亡状态。HE染色和免疫组化染色判断造釉型颅咽管瘤（ACP）和乳头型颅咽管瘤（PCP）的炎性反应情况。结果抗体芯片筛选结果显示ACP中3种炎性因子高表达，分别为单核细胞趋化蛋白1（MCP-1，7±1）、白介素6（IL-6，6±1）和白介素1p（IL-1β，8±2），且均高于PCP（分别为2±1、2±1、2±1）、下丘脑胶质瘤（分别为2±1、1．4-0、2±1）、垂体腺瘤（分别为14-0、2±1、2±1）和脑膜瘤（分别为1±0、1-t-O、1±0）（均P〈0．05）。qPCR结果证实ACP中MCP-1、IL-6、IL-1β的转录水平[分别为（180±24）pg／ml、（266±19）pg／ml、（243±31）pg／ml]均高于PCP[分别为（68±14）pg／ml、（98±29）pg／ml、（154±21）pg／ml]、下丘脑胶质瘤[分别为（60±14）pg／ml、（142±29）pg／ml、（149±39）pg／ml）]、垂体腺瘤[分别为（180±24）pg／ml、（266±19）Pg／ml、（134±27）pg／ml]和脑膜瘤[分别为（56±14）pg／ml、（89±19）pg／ml、（121±21）pg／ml）]（均P〈0．05）。ELISA结果显示，ACP培养上清中MCP-1、IL-6、IL-1β水平[分别为（168±19）pg／ml、（155±34）pg／ml、（228±31）pg／ml）]显著高于PCP[分别为（66m124）pg／ml、（89m119）pg／ml、（112±21）pg／ml）]（均P〈0．05）。CATH-a细胞被ACP上清处理后的凋亡率显著升高（P〈0．05），而PCP无显著促凋亡作用（P〉0．05）。ACP以巨噬细胞浸润的2、3级炎性反应为主，PCP以1、2级为主，ACP的炎性反应程度高于PCP（P〈0．05）。结论颅咽管瘤组织内存在炎性反应。ACP的炎性反应程度较PCP高，以MCP-1、IL-6和IL-1β升高为主，且对神经细胞具有毒性作用。

Objective To investigate the clinical significance of inflammatory reaction in craniopharyngiomas. Methods Antibody microarray was employed for detection of inflammatory cytokines expression in craniopharyngiomas and other common tumors in sellar region （including the pituitary adenoma, meningioma, pilocytic astrocytoma）. The methods of qPCR and ELISA were further used for verifying the expression of inflammatory factors in each group. At the same time, the mouse brain neuron cell line （ CATH. a） was treated with supernatant of the primary culture medium of craniopharyngiomas, and the apoptotic state of neurons was explored by flow cytometry. Results The craniopharyngioma was more likely to have an inflammatory response than other sellar tumors. The results of antibody microarray showed that the relative expression levels of monocyte chemotactic protein-1 （ MCP-1 ）, interleukin 6 （IL-6） and interleukin i （IL-1） in adamantinomatous craniopharyngiomas （ACP） （7 ± 1, 6 ± 1, 8 ± 2 ） were significantly higher than those in papillary craniopharyngiomas （PCP） （2 ± 1, 2 ± 1, 2 ± 1 ）, pilocytic astrocytomas （2 ± 1, 1 ± 0, 2 ± 1 ）, pituitary adenomas （ 1 ± 0, 2± 1,2 ± 1 ） and meningiomas （ 1 ± 0, 1 ± 0, 1 ± 0） （ all P 〈 0.05）. The qPCR results further verified that the transcriptional levels of 3 inflammatory factors mentioned above in ACP （ 180 ± 24 pg/ml, 266± 19 pg/ml, 243 ± 31 pg/ml） were significantly higher than those in PCP （68 ± 14 pg/ml, 98 ±29 pg/ml, 154 ±21 pg/ml）, pilocytic astrocytomas （60 ± 14 pg/ml, 142 ±29 pg/ml, 149 ±39 pg/ml） , pituitary adenomas （180±24 pg/ml, 266 ± 19 pg/ml, 134±27 pg/ml） and meningiomas （56 ± 14 pg/ml, 89 ± 19 pg/ml, 121 ±21 pg/ml） （all P 〈0. 05）, while Elisa results showed that three kinds of inflammatory factors in the ACP primary cell culture medium supernatant （168 ± 19 pg/ml, 155 ± 34 pg/ml, 228 ± 31 pg/ml） were significantly higher than those in PCP （66 ± 24 pg/ml, 89 ± 19 pg/ml, 112 ±21 pg/ml） （all P 〈 0.05）. After CATH. a cells were stimulated by ACP medium supernatant, their apoptosis increased significantly, while the promotion of apoptosis by supernatant of PCP medium was not obvious. Immunohistochemistry results showed that the inflammatory response in ACP tissue was characterized by infiltration of macrophage inflammatory cells; and activated mieroglias were observed in the front area between tumor and the third ventricle floor. Conclusions Inflammatory reaction could be found in the craniopharyngioma and the levels of MCP-1, IL-6 and IL-1 were higher in ACP than PCP. The inflammatory reaction of craniopharyngioma seems toxic to CATH. a cells.