IL-6和IL-6R在脑胶质瘤发病中的作用初探

目的：初步探讨ＩＬ－６和ＩＬ－６Ｒ与脑胶质瘤发生发展的关系。方法：采用逆转录聚合酶链反应（ＲＴ－ＰＣＲ）方法检测白细胞介素６（ＩＬ－６）和ＩＬ－６受体（ＩＬ－６Ｒ）基因在脑胶质瘤组织标本和正常人脑胶质细胞中的表达。结果：３０例脑胶质瘤组织标本中有２４例（８０．０％）表达ＩＬ－６、２６例（８６．７％）表达ＩＬ－６Ｒ、２２例（７３．３％）同时表达ＩＬ－６和ＩＬ－６Ｒ。人脑正常胶质细胞仅有ＩＬ－６基因弱表达,而玩ＩＬ－６Ｒ表达。结论：脑胶质瘤可能存在与肿瘤的恶性增殖有关的ＩＬ－６／ＩＬ－６Ｒ自分泌或旁分泌环路。     

IL—6在脑胶质瘤发生发展中的作用

脑胶质瘤是神经系统肿瘤中发病率和复发率最高的肿瘤，但其病因尚不十分清楚。国内外学者先后从培养的人脑胶质瘤细胞株和胚胎脑胶质细胞中观察到ＩＬ－６基因的持续表达和ＩＬ－６的持续分泌，并可在其它细胞因子的刺激下提高ＩＬ－６基因表达量，初步推测脑胶质瘤的发生发展可能与这些细胞因子基因异常表达及分泌有关。     

IL—6R与脑胶质瘤的恶性增殖

白细胞介素－６（ＩＬ－６）与ＩＬ－６受体（ＩＬ－６Ｒ）的自分泌或旁分泌环路在肿瘤发生发展中的作用越来越受到重视。随着对ＩＬ－６Ｒ结构和功能及其与脑胶质瘤关系的研究不断深入，初步推测脑胶质瘤细胞及组织中也存在ＩＬ－６－ＩＬ－６Ｒ自分泌或旁分泌环路，这为脑胶质瘤的免疫治疗提供了新的思路。     

Th2类细胞因子优势表达在人脑胶质瘤发生及发展中的作用

目的 探讨Th1／Th2类细胞因子基因在人脑胶质瘤中的分泌及表达情况,以及它们在脑胶质瘤发生及发展中的作用。方法 以IL—2、IFNγ代表Th1类细胞因子,IL—4、IL—6及IL—10代表Th2类细胞因子,采用ELISA法检测脑胶质瘤细胞株培养上清中细胞因子的活性,采用逆转录聚合酶链反应方法(RT—PCR)检测Th1／Th2类细胞因子基因在脑胶质瘤细胞、肿瘤浸润淋巴细胞及细胞株中的表达情况。结果 人脑胶质瘤细胞、肿瘤浸润淋巴细胞及脑胶质瘤细胞株均呈现明显的Th2类细胞因子基因优势表达,而正常人脑组织中则无这种表达趋势。结论 Th2类细胞因子在人脑胶质瘤中的优势表达可能是导致脑胶质瘤患者细胞免疫功能低下的原因之一,有可能在脑胶质瘤的发生及发展中起一定作用。     

人脑胶质瘤细胞中IL—6mRNA的表达及意义

目的 观察白细胞介素－６（ＩＬ－６）ｍＲＮＡ在脑胶质瘤细胞中的表达。方法 采用逆转聚合酶链反应（ＲＴ－ＰＣＲ）方法对２０例新鲜人脑胶持瘤标本进行了检测。结果 ２０例标本中有１７例表达ＩＬ－６ｍＲＮＡ，恶性程度高的肿瘤中ＩＬ－６ｍＲＮＡ的表达量明显高于恶性程度低的肿瘤。结论脑胶质瘤细胞中ＩＬ－６基因的表达与脑胶质瘤的发生发展有关。     

miR-129-5p在人脑胶质瘤组织中表达下调的表观遗传学机制

目的:检测miR-129成熟体miR-129-5p在人脑胶质瘤组织中的表达特点,探讨人脑胶质瘤组织中miR-129-2表达异常是否与miR-129-2启动子甲基化有关.方法:收集湘雅医院神经外科2013年10-12月收治的胶质瘤患者肿瘤组织21例、脑外伤患者非肿瘤脑组织标本21例.Real-time PCR检测脑胶质瘤组织和非肿瘤脑组织、脑胶质瘤细胞株和正常脑胶质细胞株中miR-129-5p的表达;甲基化特异性PCR(methylation-specific PCR,MSP)检测胶质瘤组织中miR-129-2基因启动子甲基化的情况;亚硫酸氢盐硫化测序PCR(bisulfite-sequencing PCR,BSP)分析经甲基化转移酶抑制剂5-氮杂-2'-脱氧胞苷(5-aza-2'-deoxycitydine,5-Aza-dC)处理后的胶质瘤细胞miR-129-2基因启动子甲基化水平的变化,同时Real-time PCR检测miR-129-5p的表达变化.结果:miR-129-5p在人脑胶质瘤组织和细胞中表达显著低于非肿瘤脑组织和正常脑胶质细胞(P＜0.05),胶质瘤组织中miR-129-2基因的启动子甲基化率显著高于正常脑组织(P＜0.05),使用5-Aza-dC去甲基化处理脑胶质瘤细胞后miR-129-2基因的启动子甲基化程度降低、miR-129-5p表达显著升高(P＜0.05).结论:miR-129-5p在脑胶质瘤中显著低表达,miR-129-2启动子甲基化是导致其低表达的机制之一.     

沙利度胺对骨髓瘤细胞IL-6及其传导途径的影响

目的　探讨沙利度胺治疗难治复发性多发性骨髓瘤 (MM)的有效机制。方法　用双抗夹心法 (ELISA)动态检测MM患者血清白细胞介素 6 (IL 6 )水平 ,流式细胞仪检测瘤细胞表面IL 6受体 (IL 6R)的表达 ,RT PCR半定量法检测IL 6Rβ亚单位mRNA的表达。 结果　口服 2 0 0mg/d沙利度胺前 ,MM患者血清IL 6水平为 5 6 4.8± 319.4ng/L ,瘤细胞表面IL 6R阳性率为 33.6 % ;口服 2 0 0mg/d沙利度胺后第 14天 ,IL 6水平为 5 6 0 .3± 414.8ng/L ,瘤细胞表面IL 6R阳性率为 31.8% ,分别与口服沙利度胺前比较 ,差异无显著性 (P >均 0 .0 5 )。口服 40 0mg/d沙利度胺第 14,2 8,42 ,5 6 ,84天 ,IL 6水平分别为 5 16 .7± 131.9、42 6 .7± 180 .4、387.9± 187.4、35 0 .1± 85 .5和 2 12 .3± 92 .5ng/L ,瘤细胞表面IL 6R阳性率分别为 2 8.5 %、2 4.3%、2 1.3%、12 .6 %和 10 .1% ,均分别低于口服 2 0 0mg/d沙利度胺前IL 6水平或瘤细胞表面IL 6R阳性率 (P <0 .0 5或P <0 .0 1) ;口服 2 0 0mg/d沙利度胺前及口服第 14天的IL Rβ亚单位mRNA比值分别为 7.8和 6 .9,二者差异无显著性 (P >0 .0 5 ) ;口服沙利度胺 40 0mg/d第 14,2 8天IL Rβ亚单位mRNA的比值分别为 5 .3和 2 .7,与口服 2 0 0mg/d沙利度胺前的比较 ,差异有显著性 (P     

恶性胶质瘤中IL-13Rα2和PCNA的表达及其与病理特征和预后的关系

目的：检测人脑恶性胶质瘤组织中IL13Rα2和增殖细胞核抗原（proliferating cell nuclear antigen, PCNA）的表达强度,分析其与患者临床病理特点和预后之间的关系。方法：选取20022006年温州医学院附属第一医院手术治疗的43例恶性胶质瘤标本;采用免疫组织化学法检测并用图像分析系统分析IL13Rα2和PCNA在恶性胶质瘤组织中的表达,分析两者间的相关性;分析它们与临床特征和预后的关系。结果：(1) 43例恶性胶质瘤标本有40例(93%)IL13Rα2阳性表达和36例(84%)PCNA阳性表达;(2) IL13Rα2和PCNA表达强度与肿瘤部位和肿瘤大小无相关性(P>0.05);在WHOⅢ级与Ⅳ级间两者表达强度差异均有统计学意义（P=0.031, P=0.002）;在生存时间≤6个月与>6个月患者间IL13Rα2 表达强度差异有统计学意义(P=0.028)。(3)IL13Rα2和PCNA的表达强度呈显著的正相关(r=0.653,P=0.000)。结论：人恶性胶质瘤组织中IL13Rα2和PCNA均高表达,前者表达强度与肿瘤恶性分级和患者预后密切相关,在恶性胶质瘤的诊断和预后判断中具有潜在的临床应用价值。     

联合IL-2基因和B7-1基因转染G422细胞体外生物学特性

目的　通过研究联合基因转染后胶质母细胞瘤G42 2细胞生物学特性的变化 ,探讨脑胶质瘤细胞因子基因治疗的新途径。方法　通过重组腺病毒作载体 ,将IL 2基因和B7 1基因转染至小鼠G42 2细胞 ,通过CTLL 2细胞增殖的MTT法检测IL 2的分泌水平 ,FACS检测B7 1和ICAM 1的表达 ,MTT法检测转基因后G42 2细胞的增殖活性 ,双层琼脂糖培养法检测其克隆形成能力。结果　IL 2基因转染 4小时后即可检测到IL 2的分泌 ,2 4小时达高峰 ( 4 83U/ml) ,一周时表达明显下降 ,B7 1高表达 ,ICAM 1表达增加 ,增殖能力和克隆形成能力无变化。结论　重组腺病毒作载体 ,IL 2基因及B7 1基因共转染G42 2细胞 ,能得到目的基因的高效表达 ,同时促进了粘附分子ICAM 1的表达 ,体外增殖活性不受影响。     

细胞因子基因表达与人脑胶质瘤关系的研究

目的 探讨细胞因了在脑胶质瘤发生及发展中的作用。方法 采用逆转录聚合酶链反应(RT—PCR)方法测定五种细胞因子基因在人脑胶质瘤中的表达。结果 表明,介导体液免疫的细胞因子(IL—4、IL—6、IL—10)基因呈优势表达,而介导细胞免疫的细胞因子(IL—2、IFNγ)基因呈劣势表达。结论 两类细胞因子在人脑胶质瘤中的漂移可能是造成脑胶质瘤患者细胞免疫功能低下的原因之一。     

自泌性白细胞介素—6对人肺腺癌细胞系PAa生长的影响

目的　探讨白细胞介素 - 6( interleukin- 6,IL- 6)对人肺癌细胞体外生长的影响。方法　应用逆转录 PCR技术检测人肺腺癌细胞系 PAa IL- 6受体 m RNA、Northern杂交技术检测 PAa细胞IL- 6m RNA;用生物活性检测法测定 PAa细胞所分泌的 IL- 6,并用抗体中和法检测内源性 IL- 6对PAa细胞体外生长的影响。结果　PAa细胞表达 IL- 6受体 m RNA,重组人 IL- 6刺激 PAa细胞生长 ;PAa细胞表达 IL- 6m RNA且分泌有生物活性的 IL- 6,抗 IL- 6抗体抑制 PAa细胞生长。结论　 IL- 6对 PAa细胞生长具有调控作用 ,IL- 6为 PAa细胞自分泌生长刺激因子。     

A dialog between glioma and microglia that promotes tumor invasiveness through the CCL2/CCR2/interleukin-6 axis

Glioma cells in situ are surrounded by microglia, suggesting the potential of glioma-microglia interactions to produce various outcomes. As chemokines are important mediators of cell-cell communication, we sought first to identify commonly expressed chemokines in 16 human glioma lines. We found CCL2 (macrophage chemoattractant protein-1) messenger RNA to be expressed by the majority of glioma lines. However, these lines did not express the CCL2 receptor, CCR2, which was found on microglia. Next, we overexpressed CCL2 in the U87 glioma line, which has low basal level of CCL2, to investigate the hypothesis that glioma-secreted CCL2 interacts with microglia to affect glioma growth. Stable clones with 10- to 12-fold elevation of CCL2 have similar growth rate and invasive capacity as vector controls when cultured in isolation. However, in coculture with microglia in a three-dimensional collagen gel matrix, the invasiveness of CCL2-overexpressing clones was increased. Gene array analyses were then undertaken and they revealed that interleukin (IL)-6 was consistently increased in the coculture. Recombinant IL-6 enhanced the invasiveness of glioma cells when these were cultured alone, whereas a neutralizing antibody to IL-6 attenuated the microglia-stimulated glioma invasiveness. Finally, we found that human glioma specimens in situ contained IL-6 immunoreactivity that was expressed on CD68+ cells. This study has uncovered a mechanism by which glioma cells exploit microglia for increased invasiveness. Specifically, glioma-derived CCL2 acts upon CCR2-bearing microglia, which then produces IL-6 to stimulate gliomas. The CCL2/CCR2/IL-6 loop is a potential therapeutic target for the currently incurable malignant gliomas.     

Autocrine glutamate signaling promotes glioma cell invasion

Malignant gliomas have been shown to release glutamate, which kills surrounding brain cells, creating room for tumor expansion. This glutamate release occurs primarily via system xC, a Na+-independent cystine-glutamate exchanger. We show here, in addition, that the released glutamate acts as an essential autocrine/paracrine signal that promotes cell invasion. Specifically, chemotactic invasion and scrape motility assays each show dose-dependent inhibition of cell migration when glutamate release was inhibited using either S-(4)-CPG or sulfasalazine, both potent blockers of system xC. This inhibition could be overcome by the addition of exogenous glutamate (100 micromol/L) in the continued presence of the inhibitors. Migration/invasion was also inhibited when Ca2+-permeable alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptors (AMPA-R) were blocked using GYKI or Joro spider toxin, whereas CNQX was ineffective. Ca2+ imaging experiments show that the released glutamate activates Ca2+-permeable AMPA-R and induces intracellular Ca2+ oscillations that are essential for cell migration. Importantly, glioma cells release glutamate in sufficient quantities to activate AMPA-Rs on themselves or neighboring cells, thus acting in an autocrine and/or paracrine fashion. System xC and the appropriate AMPA-R subunits are expressed in all glioma cell lines, patient-derived glioma cells, and acute patient biopsies investigated. Furthermore, animal studies in which human gliomas were xenographed into scid mice show that chronic inhibition of system xC-mediated glutamate release leads to smaller and less invasive tumors compared with saline-treated controls. These data suggest that glioma invasion is effectively disrupted by inhibiting an autocrine glutamate signaling loop with a clinically approved candidate drug, sulfasalazine, already in hand.     

白介素-6 对肺癌诊断及预后评价的临床价值

 目的　探讨血清白介素 - 6(IL- 6)水平测定对肺癌诊断以及远道转移之间关系。方法　采用放免方法测定 44例不同组织类型肺癌与 34例健康对照组血清中白介素 - 6水平。结果　肺癌组患者 IL- 6水平显著高于对照组 (P<0 .0 1 )。不同组织来源肺癌组之间 IL- 6变化无显著性差异(P>0 .0 5) ,有远道转移组 (M1组 )较无转移组 (M0 组 ) IL- 6水平显著下降 (P<0 .0 5)。结论　测定血清中 IL- 6水平对肺癌的诊断 ,有无远道转移及预后评估均有重要临床意义。     

IN VIVO GROWTH OF CEREBRAL C6 GLIOMA CELLS TRANSFECTED AND TREATED WITH CX43 GENE

Objective: To study the role of connexin gene (Cx43) on the development of glioma and the feasibility of using Cx43cDNA as a target of gene therapy of gliomas.Methods: Parental rat C6 cells and C6 cells transfected with Cx43cDNA were implanted into right caudate nucleus of SD rats as control and transfected group.Rats bearing cerebral C6 gliomas were treated with Cx43cDNA and empty vector as treated group and empty vector group. The general manifestation, survival time, MRI dynamic scanning and histopathological changes of all rats were observed. In situ hybridization and immunohisto- chemistry were used for examination of Cx43mRNA and its protein in gliomas. Average number of AgNOR staining was used for detection of cell proliferation activity, and TUNEL method for determination of cell apoptosis. Results: All rats in control and empty vector group died of cerebral gliomas within 3 weeks after implantation of C6 cells. Six out of nine rats in the transfected group and eight out of ten rats in treated group kept alive beyond 120 days with totally disappearing of the tumor foci, except one treated rat having a little residue of tumor. In gliomas of transfected and treated groups Cx43 gene expression was upregulated, proliferation activity was lowered,However, the apoptotic cells did not increase.Conclusion: The present study indicates that Cx43 gene is of crucial importance in the development of malignant glioma. It can be an effective target for gene therapy of gliomas.