IFNγ和IL-6或IL-1联合应用对人肝癌细胞系C1抑制物合成的作用

为研究细胞因子联合作用时对人肝癌细胞系Ｃ１抑制物（Ｃ１ＩＮＨ）合成的影响，用ＥＬＩＳＡ 双夹心法测定了经细胞因子联合作用后ＨｅｐＧ２和ＳＭＭＣ７７１２细胞培养上清中Ｃ１ＩＮＨ的含量。结果表明：ＩＦＮγ分别与ＩＬ－６和ＩＬ－１共同作用时，能显著增加人肝癌细胞系Ｃ１ＩＮＨ的合成，其作用强于同剂量ＩＦＮγ的单独作用。ＩＬ－１本身不能增加ＨｅｐＧ２细胞Ｃ１ＩＮＨ的合成。ＩＬ－１对ＩＬ－６的上调没有增强作用，也无抑制作用。     

IFNγ和IL—6对人肝癌细胞系C1抑制物合成的调控

为研究ＩＦＮγ或ＩＬ－６对肝癌细胞系Ｃ１抑制物合成的调控作用。方法用ＥＬＩＳＡ双夹心法检测了经其刺激后ＨｅｐＧ２和ＳＭＭＣ７７１２细胞培养上清中Ｃ１ＩＮＨ的含量。结论此结果提示有可能用ＩＦＮγ和ＩＬ－６治疗Ｃ１ＩＮＨ缺乏病。     

IFN—γ抑制血管成形术后兔血管平滑肌细胞增殖和PDGF—A的表达

目的和方法：采用体外细胞培养，进行氚标胸腺嘧啶核苷（３Ｈ－ＴｄＲ）掺入和原位杂交。结果：发现γ－干扰素（ＩＦＮ－γ）能抑制体外培养新西兰兔血管平滑肌细胞（ＶＳＭＣ）的ＤＮＡ合成和血小板衍化生长因子Ａ链（ＰＤＧＦ－Ａ）的基因表达；整体实验中，用４ＦＦｏｇａｒｔｙ导管气囊拉伤新西兰兔的腹主动脉内皮后，随即连续肌注ＩＦＮ－γ（１６５万Ｕ·ｋｇ１·ｄ１）３天，结果发现肌注ＩＦＮ－γ能明显抑制受损部位ＶＳＭＣ的３Ｈ－ＴｄＲ的标记率及ＰＤＧＦ－Ａ的表达。结论：ＩＦＮ－γ对血管成形术引起的血管挛缩有一定的舒张作用。     

人中性粒细胞抑制吞噬细胞生成肿瘤坏死因子机制的研究

目的：初步探讨人中性粒细胞（ＰＭＮ）抑制Ｕ９３７细胞肿瘤坏死因子－α（ＴＮＦ－α）生成的机制。方法：将人ＰＭＮ与经ＰＭＡ刺激分化为单核细胞样细胞的Ｕ９３７细胞在不同条件下共同培养,取上清测定ＴＮＦ－α含量。结果Ｌ：ＰＭＮ与Ｕ９３７细胞共同培养可抑制Ｕ９３７细胞生成ＴＮＦα。ＰＭＮ的抑制作用可被代剂量叠氮钠逆转达４５％。该剂量叠氮钠并不影响Ｕ９３７细胞产生ＴＮＦ－α。叠氮钠的作用机制并非抑制了ＰＭＮ     

dt2—cAMP,IFN—γ和PMA对U937胞浆Ca^2＋和酪氨酸激？…

目的 探讨ｄｔ２－ｃＡＭＰ，ＰＭＡ和ＩＦＮ－γ对Ｕ９３７细胞Ｃ５ａＲ表达的影响，以及ｒｈＣ５ａ刺激受ｄｔ２－ｃＡＭＰ，ＰＭＡ和ＩＦＮ－γ分化的Ｕ９３７细胞后，其胞浆Ｃａ＾２＋浓度的变化情况。方法 用流式细胞仪分析胞膜Ｃ５ａＲ和细浆蛋白酪氨酸激酶的表达；荧光发光法测定胞浆Ｃａ＾２＋浓度的变化。结果 ｄｔ２－ｃＡＭＰ，ＰＭＡ和ＩＦＮ－γ等可不同程度地上调Ｃ５ａＲ表达。用０．５ｍｍｏｌ／Ｌ的ｄｔ２－ｃＡ     

rTNF_(-α)、IFN_(γ)对人脐静脉内皮细胞表达粘附分子的影响

为了研究细胞因子ｒＴＮＦ α、ＩＦＮγ对内皮细胞表达粘附分子的影响,及ｒＩＦＮγ对ＴＮＦ α的协同效应。采用ｒＴＮＦ α、ＩＦＮγ诱导培养的人脐静脉内皮细胞（ＨＵＶＥＣ）,以Ｃｅｌ ＥＬＩＳＡ法检测粘附分子ＩＣＡＭ １、ＥＬＡＭ １和ＶＣＡＭ １的表达。结果表明,ｒＴＮＦ α以浓度和时间依赖的方式诱导ＨＵＶＥＣ表达ＩＣＡＭ １、ＥＬＡＭ １和ＶＣＡＭ １,ＩＦＮγ促进ｒＴＮＦ α诱导的ＶＣＡＭ １表达。ＩＦＮ α对ｒＴＮＦ α诱导效应无影响。提示ｒＴＮＦ α可诱导ＨＵＶＥＣ表达粘附分子,而ＩＦＮγ可促进ｒＴＮＦ α诱导的ＨＵＶＥＣ表达ＶＣＡＭ １。     

γ－干扰素在淋巴细胞介导的子宫颈角化上皮细胞毒作用中的意义

本研究以子宫颈角化上皮为靶细胞，用试管培养技术及非同位素细胞毒性试验，探讨了干扰素（ＩＦＮ－γ）对淋巴细胞介导的对宫颈角化上皮细胞毒作用的影响及细胞间吸附分子１（ＩＣＡＭ－１）与白细胞功能相关抗原１（ＬＦＡ－１）在细胞毒作用中的意义。结果显示ＣａＳｋｉ、ＳｉＨａ、ＨｅＬａ、Ｗ１２及ＮＣｘ均为ＮＫ抵抗、ＬＡＫ敏感细胞。ＩＦＮ－γ预处理靶细胞明显增加ＬＡＫ细胞的杀伤活性，但对ＮＫ活性影响不大；ＩＦＮ－γ预处理效应细胞可增强ＬＡＫ细胞对上述靶细胞的溶解作用，但并不改变ＮＫ对靶细胞的选择性；抗ＩＣＡＭ－１与ＬＦＡ－１单克隆抗体能有效地降低ＬＡＫ细胞对靶细胞的杀伤作用，而抗ＨＬＡＡＢＣ及ＨＬＡＤＲ则无此作用。提示ＩＦＮ－γ仅对ＬＡＫ细胞介导的对宫颈角化上皮的细胞毒作用有明显增强作用，而对ＮＫ细胞无明显影响；ＩＣＡＭ－１－ＬＦＡ－１通路为ＬＡＫ细胞结合并溶解靶细胞的主要通路，且这种细胞毒作用是非ＭＨＣ限制的。     

γ—干扰素在淋巴细胞介导的子宫颈角化上皮细胞毒…

本研究以子宫颈角化上皮为靶细胞，用试管培养技术及非同位素细胞毒性试验，探讨了干扰素（ＩＦＮ－γ）对淋巴细胞介导的对宫颈角化上皮细胞毒作用的影响及细胞间吸附分子１（ＩＣＡＭ－１）与白细胞功能相关抗原１（ＬＦＡ－１）在细胞毒作用中的意义。结果显示ＣａＳｋｉ、Ｓｉｈａ、ＨｅＬａ、Ｗ１２及ＮＣｘ均为ＮＫ抵抗、ＬＡＫ敏感细胞。ＩＦＮ－γ预处理靶细胞明显增加ＬＡＫ细胞的杀伤活性，但对ＮＫ活性影响不大；ＩＦＮ－     

DEFINITION AND MECHANISM RESEARCH OF NK CELLS AC-TIVATION THROUGH THE CD45 PATHWAY

为鉴定介导ＮＫ细胞激活的表面分子及其作用机制，本研究用一系列粘附分子单抗及配体体外刺激ＮＫ细胞，定量检测ＮＫ细胞ＩＦＮ－γ的分泌；检测了ＮＫ细胞经ＩＬ－２诱导后其异构体的变化及其不同异构体单抗对ＮＫ细胞的刺激作用；以ＣＤ２２α、ＣＤ２２β基因导入Ｂ７８Ｈ１细胞后观察转染细胞对ＮＫ细胞的粘附和刺激效应。结果表明：ＩｇＧ２ａ和ＩｇＧ１亚类ＣＤ４５单抗及其Ｆ（ａｂ）２均可独立刺激ＮＫ细胞释放ＩＦＮ－γ；ＮＫ细胞经ＩＬ－２培养诱导后其表面ＣＤ４５分子异构体的表达呈ＲＯ逐渐增加，而ＲＡ逐渐减少的特征；ＣＤ４５ＲＯ单抗可刺激ＮＫ细胞释放ＩＦＮ－γ；ＣＤ２２α、ＣＤ２２β转染细胞可粘附，但不能刺激ＮＫ细胞激活。结果提示：ＮＫ细胞可经ＣＤ４５ＲＯ途径激活并释放ＩＦＮ－γ，该途径与ＣＤ１６分子无关，ＣＤ４５分子的特异性配体并非既往所报道的ＣＤ２２，尚待进一步鉴定     

人C_1q分子对Raji、U_（937）细胞产生IL－1活性的抑制调节

为探讨Ｃ_１ｑ与免疫细胞产生的ＩＬ－１活性的关系，采用Ｒａｊｉ、Ｕ_（９３７）及对照ＭｏｌＴ_４三株培养细胞与Ｃ_１ｑ共同孵育２０ｈ后收集细胞上清，以３~Ｈ－ＴｄＲ掺入法检测ＩＬ－１活性，结果显示Ｃ_１ｑ能明显抑制Ｒａｊｉ及Ｕ_（９３７）细胞ＩＬ－１活性，对ＭｏｌＴ_４细胞无此作用，Ｆ（ａｂ＇）_２ａｎｔｉＣ_１ｑ可阻断此抑制作用；初步探讨其作用机制，推测可能Ｃ_１ｑ通过Ｃ_１ｑ受体刺激该细胞产生ＩＬ－１ＩＮＨ而抑制ＩＬ－１活性。     

人食管癌相关核基质蛋白的初步研究

目的 从核基质蛋白中探寻人食管癌肿瘤标志物。方法 用高盐抽提法制备核基质,免疫动物获取抗血清,Western Blot检测特异性。结果 食管癌细胞中出现一条阳性条带,其代表的蛋白约为126000u,除食管癌原发的贲门癌外,这一阳性条带均未在正常食管和所检测的其它几种病细胞中出现。结论 食管癌细胞核基质中存在1个或1组表观分子量为126000u的相关多肽,可能与食管癌的发生发展有关,并且有一定的诊断价值。     

核基质蛋白在肿瘤研究中的应用

核基质蛋白(nuclear matrix proteins,NMPs)是一种参与细胞核构建、染色质/体构成、DNA复制和转录的具有组织特异性和肿瘤相关性的重要蛋白质。目前在膀胱癌、前列腺癌、食管癌、白血病、肝癌等肿瘤组织细胞中均发现了NMPs的特异性表达和变化。近年来,NMPs在肿瘤发生、发展中的作用,在肿瘤的诊断、治疗中的价值成为人们研究的热点。     

核基质与肿瘤研究进展

核基质又称核骨架,是指真核细胞的核外周核纤层蛋白、核孔复合物、内部核蛋白网络以及残余的核仁等核内网架结构。核基质与DNA的复制、RNA的转录与修饰及染色体的组装等许多重要生命活动相关。在多种肿瘤中相继发现肿瘤相关性核基质蛋白。本文将对核基质在肿瘤的发生、演进和转移中的作用,及其在肿瘤的诊断、治疗中的价值作一综述。     

The Aberrant Expressions of Nuclear Matrix Proteins During the Apoptosis of Human Osteosarcoma Cells

The objective of this study was to investigate altered expressions of nuclear matrix proteins (NMPs) of human osteosarcoma (OS) MG‐63 cells during curcumin‐induced apoptosis of human OS MG‐63 cells. MG‐63 cells were cultured with curcumin (7.5 mg/L) for 72 hr. Morphological alterations of cells were captured using light microscopy and transmission electron microscopy, and cell cycle distribution was estimated by flow cytometry. NMPs were selectively extracted and subjected to two‐dimensional gel electrophoresis (2‐DE) analysis. Western blots were performed to determine changes in the expression levels of specific NMPs. The results demonstrated that typical characteristics of apoptosis were observed. Cellular chromatin agglutinated, cell nuclei condensed, and apoptotic bodies were formed after treatment with curcumin. The 2‐DE results displayed 27 NMPs, 21 of which were identified to have change in expression levels significantly during apoptosis. The altered expressions of three of these NMPs (nucleophosmin, prohibitin, and vimentin) were further confirmed by immunoblotting. These findings indicated that the apoptosis of MG‐63 cells was accompanied by the expression alteration of NMPs. Our results might help to reveal the relationship between NMPs and the regulation of gene expression in the process of apoptosis, as well as provide the basic concepts for future studies on the mechanisms of apoptosis and the therapy for bone diseases. Anat Rec, 2010. © 2010 Wiley‐Liss, Inc.     

伪狂犬病病毒Ea株诱导牛肾细胞凋亡及核基质蛋白的变化

目的研究伪狂犬病病毒(PrV)是否具有诱导组织培养细胞发生细胞凋亡的功能及探讨凋亡细胞核基质蛋白表达的变化。方法利用锥虫蓝(台盼蓝)染色绘制PrV在不同细胞上的增殖曲线;荧光染色观察凋亡细胞核的形态特征;琼脂糖凝胶电泳分析细胞染色体DNA的片段化;高分辨率双向电泳分析细胞凋亡前后核基质蛋白的表达差异。结果膜通透性的DNA特异性结合染料Hoechst 33342染色显示,PrV感染后36h,牛肾(MDBK)细胞核染色质开始固缩、凝聚,细胞核碎裂;DNA提取及琼脂糖凝胶电泳分析表明,细胞染色体DNA发生片段化,形成“DNA梯状”条带(DNA ladder);尽管PrV可以诱导MDBK细胞发生细胞凋亡,但不能诱导IBRS-2,BHK-21及PK-15细胞发生凋亡。MDBK细胞发生凋亡前后核基质蛋白的表达发生改变,存在表达上调、下调、诱导表达及表达遏制等情况。结论PrV诱导细胞发生细胞凋亡是其致细胞死亡的形式之一,间接反映了PrV与宿主细胞间复杂的相互作用。细胞凋亡前后核基质蛋白表达存在的差异说明PrV致MDBK细胞发生凋亡是自身基因产物或诱导或抑制宿主细胞基因表达的结果。     

姜黄素诱导人食管癌EC9706细胞凋亡过程中核磷蛋白的表达与定位变化

目的探讨核磷蛋白NPM在癌细胞诱导凋亡过程中在细胞内、细胞核基质上的定位与表达变化,以及NPM与凋亡调控相关蛋白的关系,探索其在凋亡调控中的作用。方法在姜黄素诱导人食管癌EC9706细胞凋亡的基础上,以亚细胞蛋白质组学方法分析NPM在核基质中的存在与变化,并以免疫印迹法杂交实验进行确证;激光扫描共焦显微镜观察NPM在EC9706细胞凋亡过程中的定位与变化,以及NPM与Bax、Bcl-2等基因产物的共定位关系。结果 NPM存在于EC9706细胞核基质蛋白组分中,并在姜黄素处理后表达下调。NPM在EC9706细胞凋亡过程中发生显著的胞质-核之间的穿梭定位变化,并与Bax、Bcl-2等蛋白具有共定位关系,且共定位区域发生了变化。结论 NPM是一种核基质结合蛋白,在EC9706细胞凋亡中的表达与定位变化,及其与凋亡调控蛋白的共定位关系提示,它在EC9706细胞凋亡调控中具有重要作用。     

人食管癌细胞核基质的研究

目的　研究人食管癌细胞株 EC1和 EC18的核基质形态及蛋白成分。　方法　应用细胞的选择性抽提、DGD包埋 -去包埋剂电镜制样观察核基质形态 ,SDS-聚丙烯酰胺凝胶电泳、双向电泳和免疫印迹等分析核基质蛋白成分。　结果　抽提后 ,两种细胞内均存在精细的核基质 -核纤层 -中间丝网架。低分化的 EC1细胞核基质较高分化的 EC18更细密。核基质蛋白电泳显示 ,两株细胞有数种相同的核基质蛋白成分 ,也有各自的特异性核基质蛋白。在免疫印迹分析中 ,使用抗人 Nu MA单克隆抗体检测到两株食管癌细胞中都存在 Nu MA蛋白。　结论　在食管癌细胞中 ,存在特异的核基质蛋白 ,核基质 -核纤层 -中间丝形成一个连续的体系 ,对维持细胞核的完整性和细胞功能起重要作用。     

Morphological and biochemical analysis of anti-nuclear matrix protein antibodies in human sera

Autoimmune sera have been used in the diagnosis of autoimmune diseases as well as the analysis of nuclear substructures. In an attempt to study the biological characteristics of the nuclear matrix, we screened human sera using immunofluorescent staining and immunoblot. We detected antibodies against nuclear matrix (NM), a remnant nonchromatin protein compartment after the treatment of detergent, salt and nuclease, in 212 out of 284 tested sera (74.6%) by immunoblot. Peptides with molecular weights of 70 kDa, 50 kDa and 25 kDa were detected in the order of frequency. Clinical informations of 198 out of 212 cases were available and went as follows: 38 cases were autoimmune diseases, such as systemic lupus erythematosus and rheumatoid arthritis; 132 non-autoimmune and non-neoplastic diseases; 16 neoplastic diseases and 12 cases unclassified. The immunofluorescent staining intensity by anti-nuclear matrix protein (NMP) antibodies decreased variably, but fibrillogranular, speckled and nucleolar immunolocalization patterns were retained after in situ fractionation. Ku70 and La protein were detected by anti-NMP antibodies. Immunolocalization by anti-NMP antibodies indicates that the NMPs constitute a variety of characteristic nuclear substructures and may serve as autoantigens in diverse human diseases. In addition, the presence of Ku70 and La protein as NMPs suggests that the NM can be functionally active in association with DNA or RNA.     

Anti-nuclear matrix antibodies in mixed connective tissue disease

Purified serum antibodies of patients suffering from mixed connective tissue disease were tested for their immunological specificity against nuclear constituents of HeLa S3 cells. In the indirect immunofluorescent staining technique, using cells and nuclei as targets, a typical speckled intranuclear staining pattern was obtained, that persisted after degradation and extraction of all nucleic acids and their associated proteins. This treatment of nuclei with detergents, DNase, RNase and high salt concentrations leave intact only the so-called nuclear matrix which is an intranuclear proteinaceous network. Further proof that nuclear matrix proteins were targets of the autoimmune reaction was obtained after separation of these proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis and electrophoretic transfer to nitrocellulose (blotting). A specific number of blot-transferred matrix proteins reacted with purified serum antibodies of 10 patients with mixed connective tissue disease, whereas this reaction was negative with normal healthy individuals. IgG preparations of 7 patients with systemic lupus erythematosus showed a weak, if any, reaction with matrix constituents. Obviously, in some connective tissue diseases serum antibodies are expressed which are directed to specific nuclear matrix antigens.