孕激素对绒癌细胞系Galectin-3表达的影响

目的通过体外试验研究孕激素对BeWo绒癌细胞系中Galectin-3蛋白的表达,探讨其对滋养细胞侵袭性相关基因表达的影响作用。方法体外培养BeWo绒癌细胞,分别加入不同浓度的孕激素24h后,采用免疫印迹技术、免疫荧光技术测定各种条件下的Galectin-3蛋白的表达。结果Bewo绒癌细胞系表达Galectin-3。分别用0、10-7、10-6、10-5mol/L孕激素处理24h后,10-7、10-6mol/L的孕激素可以显著提高BeWo绒癌细胞系中Galectin-3蛋白的表达(P<0.05,相反10-5mol/LP使Galectin-3表达降低(P<0.05)。结论(1)BeWo绒癌细胞系有Galectin-3的表达。(2)低浓度的孕激素可以诱导Galectin-3蛋白的表达。(3)高浓度的孕激素对Galectin-3蛋白的表达有下调作用。     

HLA expression in hepatocellular carcinoma cell lines

The present study undertook to investigate the biological significance of human leucocyte antigen expression in hepatocellular carcinoma and to elucidate the role of potential modulating agents on human leucocyte antigen expression. These studies used several hepatic tumour-derived cell lines as in vitro model systems. The cell lines included PLC/PRF/5 (Alexander cell line), Hep3B. HepG2, TONG PHC, HA22T/VGH, HA59T/VGH and Mahlavu, The cell lines K562 and Raji were used as negative and positive controls, respectively. K562, a B tymphoid-derived cell line, was shown to express negligible amounts of human leucocyte anligens, while Raji, an erythromyeloid-derived cell line, expressed both class I and class II human leucocyte anligens as well as their respective invariant chains, β₂-niicroglobulin and Ii. Using an ELISA, experiments performed on these cell lines confirmed the natural expression of class I and class II antigens by the HA22T/VGH and HA59T/VGH cell lines, whereas PLC/PRF/5 displayed class II surface antigens only. The effects of modulating agents such as interferon-gamma sodium bulyrate and clofazimine on human leucocyte antigen expression were investigated using the HA22T/VGH, HA59T/VGH and TONG PHC cell lines. These agents increased class 1 and class II human leucocyte antigen expression on HA22T/VGH and TONG PHC cells, but had no effect on the HA59T/VGH cell line. The results suggest a potential use for these agents as modulators of human leucocyte antigen expression by human heptocellular cell lines.     

EB病毒潜在膜蛋白对鼻咽癌细胞系CNE1生长及HLA表达的影响

目的研究ＥＢＶ－ＬＭＰ对高分化鼻咽癌细胞株ＣＮＥ１生长及ＨＬＡ表达的影响。方法以人高分化鼻咽癌细胞株（ＣＮＥ１）为对象，采用电穿孔基因转染技术，将重组ＥＢＶ－ＬＭＰ表达质粒ｐＣＭＶａ－ＬＭＰ转染ＣＮＥ１细胞。用细胞体外增殖试验、免疫组化和流式细胞术等方法，观察细胞生长及ＨＬＡ表达的变化。结果ＥＢＶ－ＬＭＰ在体外可明显促进ＣＮＥ１细胞的增殖，增殖吸光度（Ａ）比值试验组与空白组及阴性对照组比较有显著性差异（Ｐ＜０．０１）；实验组细胞软琼脂克隆形成率显著高于空白及阴性对照组（Ｐ＜０．０１）；细胞ＤＮＡ含量明显增高（Ｐ＜０．０１／０．０５）；ＦＣＭ法测定细胞角蛋白表达，实验组比空白及阴性对照组阳性率显著降低（Ｐ＜０．０５）；ＨＬＡ免疫组化检测结果表明，ＬＭＰ表达细胞系ＨＬＡⅠ类及Ⅱ类抗原的表达都明显下降（Ｐ＜０．０１）。结论ＥＢＶ－ＬＭＰ对ＣＮＥ１细胞生长有明显的促进作用且可明显抑制细胞分化，ＬＭＰ可致鼻咽癌细胞ＨＬＡ抗原的表达改变。     

hCG对JEG-3滋养细胞系表达细胞因子的影响

目的：揭示人类绒毛膜促性腺激素（hCG)是否通过改变细胞因子的生成而影响滋养细胞的侵袭性。方法：以永生化的滋养细胞系JEG－3为研究对象,采用逆转录多聚酶链反应（RT－PCR）方法观察了hCG对JEG－3细胞与细胞侵袭力调节有关的多种因素因子表达的影响,结果：JEG－3细胞表达HGF,IGF－II,VEGF和TGF－β3,且VEGF的表达以VEGF121和VEGF165为主,而不表达IGF,TGF－1β,TGF－β2,IL－β1,25U／mL hCG处理50h可显著降低JEG－3细胞中HGF的表达,同时强烈诱导VEGF121和VEGF165的表达,而其它基因的表达未发生明显变化,结论：HGF对滋养细胞的侵入起促进作用,而VEGF则具有抑制效应,说明,高浓度的hCG可能通过两种细胞因子的自分泌机制对滋养细胞的侵入起抑制作用。     

Human alloreactive CTL interactions with gliomas and with those having upregulated HLA expression from exogenous IFN-gamma or IFN-gamma gene modification.

By flow cytometry, a panel of 18 primary glioma cell explants exhibited high expression of class I HLA-A, B, C, but class II HLA-DR expression was absent. Freshly isolated normal brain cells displayed little or no HLA antigens. Alloreactive cytotoxic T lymphocytes (aCTL), sensitized to the HLA of the patient, were generated in a one-way mixed lymphocyte response (MLR). The specificity of aCTL was confirmed to be to target cells (patient glioma cells or lymphoblasts) expressing the relevant HLA antigens. However, nontumor patient-specific aCTL did not lyse normal brain cells. Titration of antibodies to HLA class I into cytotoxicity assays blocked lysis of gliomas by aCTL, confirming aCTL T cell receptor (TCR) interactions with the class I antigen on gliomas. Furthermore, aCTL interactions with glioma cells caused their apoptosis. Coincubations of aCTL with gliomas resulted in upregulated cytokine secretion. Importantly, dexamethasone, an immunosuppressive steroid used for brain edema, did not affect aCTL lytic function against tumor, indicating that steroid-dependent patients may benefit from the immunotherapy. We also explored the use of interferon-gamma (IFN-gamma) to increase aCTL tumor recognition. Coincubation of gliomas with exogenous IFN-gamma (500 U/ml, 48 h) caused a 3-fold upregulation of HLA class I and a slight induction of class II antigen expression. Gene-modified glioma cells producing IFN-gamma similarly displayed upregulated HLA expression. Glioma cells incubated with exogenous IFN-gamma or IFN-gamma-transduced glioma cells were more susceptible to lysis by aCTL than their parental counterparts, thus supporting the concept of combining IFN-gamma cytokine gene therapy with adoptive aCTL immunotherapy for brain tumor treatment.     

CoCl2对胰腺癌PC-3细胞系VEGF表达的影响

资料表明,多种肿瘤中存在着VEGF的过度表达,并且其与多种肿瘤的恶性程度及预后不良密切相关。肿瘤细胞的迅速增殖所造成的局部氧分压和pH值的改变,可能对促进VEGF等血管生成因子的分泌,启动新生血管生成发挥着重要作用。本实验的目的就在于通过CoCl2诱导肿瘤细胞低氧环境,观寨缺氧对细胞VEGF表达的影响。     

阻断VEGFR-3表达对肿瘤细胞诱导的淋巴管内皮细胞增殖的影响

目的观察阻断VEGFR-3(F lt 4)表达对肿瘤细胞(前列腺癌细胞株PC3)诱导的淋巴管内皮细胞增殖的影响。方法实验分4组,第1组为对照组。每组有6孔,每孔具有相同细胞数2×105/m l,实验组每孔各加入试剂100 m l/L。第1组(对照组)加入淋巴管内皮细胞完全条件培养液(LEC)1 m l,第2组加入LEC 1 m l+兔血清100μl,第3组加入LEC 1 m l+PC3细胞上清100μl。第4组加入LEC+抗F lt 4抗体100μl。并于24、48、72、96 h观察各组淋巴管内皮细胞生长情况。各时间段计数第1~3组细胞数,第4组于72 h计数后,去除抗体,重新加入LEC+PC3细胞上清继续培养至120 h;除96 h外,其余各时间段均计数细胞数。比较各组细胞增殖情况。结果第1、2组淋巴管内皮细胞在加试剂后各时间段两组细胞数及形态无明显差别,第3组加入PC3细胞上清后,细胞数明显多于第1、2组。第4组加试剂后24、48、72 h细胞计数均少于前24 h。清除抗体后加入PC3细胞上清48 h计数细胞,细胞数仅略见增加。结论VEGF-C高表达的PC3细胞上清能显著刺激淋巴管内皮细胞增殖,阻断F lt4表达,可在一定程度上阻断PC3细胞上清促淋巴管内皮细胞增殖作用。     

人绒毛膜促性腺激素诱导JEG-3细胞TGF-β3的表达

目的:探讨人绒毛膜促性腺激素（hCG）对绒癌细胞系表达转化生长因子（TGF-β3）的影响。方法:以绒癌细胞来源的JEG-3细胞系为研究对象,采用半定量逆转录多聚酶链反应（RT-PCR）方法,观察HCG对JEG-3细胞表达TGF-β3　mRNA的影响响。结果:分别用0、50、500、5000、25000 mU/ml　hCG处理48h后,JEG-3细胞中TGF-β3　mRNA的含量逐渐被诱导增多,并表现出一定的浓度效应——随hCG作用浓度增高而显著;另外,分别检测受25 000mU/ml hCG处理0、6、12、24、30、36、48h的JEG-3细胞中TGF-β3 mRNA的表达情况,发现TGF-β3表达受hCG的诱导在30h时最强,之后逐渐减弱。结论:hCG可能通过调节TGF-β3在滋养细胞或滋养细胞来源的绒癌细胞中的表达,进而影响细胞的浸润转移。     

Effect of 5-azacytidine on ribosomal gene expression in african green monkey ramt cell line

Institute of Medical Genetics, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. P. Bochkov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 108, No. 11, pp. 606–608, November, 1989.     

Effect of HLA phenotype and gender on expression of various forms of class I HLA in plasma.

To understand the complexity of plasma HLA antigens, the distribution of different molecular weight forms of class I HLA in plasma was investigated in 44 HLA-phenotyped and unrelated individuals. Plasma class I HLA were immunoprecipitated by using the W6/32 anti-HLA monoclonal antibody, separated by SDS-polyacrylamide gel electrophoresis and characterized by immunoblotting with the HC-10 monoclonal antibody. Four different forms of HLA heavy chains (HLA-HC) with relative molecular masses of 44, 39, 36, and 34 kd were detected. Plasma samples from all individuals contained 44 and 36 kd HLA-HC, but varied as to the presence of 39 and 34 kd HLA-HC. Eighteen percent of the individuals did not have any detectable class I HLA with 39-kd heavy chains in their plasma and 61% did not have plasma class I HLA with 34-kd heavy chains. Thus, four different distribution patterns were identified for plasma class I HLA among all individuals included in our study. The distribution patterns in four different individuals were evaluated quarterly and remained unchanged during 1 year follow-up. A significant association of absence of 39-kd plasma class I HLA-HC with female gender (p less than 0.05) and HLA-B7 phenotype (p less than 0.00015) was also found. Further pedigree analyses of four families of HLA-B7-positive and 39-kd HLA-HC-negative probands indicated that genetic factor(s) other than those associated with HLA-B7 allele and female gender is involved in regulating the expression of the plasma class I HLA with 39-kd heavy chains.     

HLA antigen expression in enteropathy associated T cell lymphoma.

AIMS: To investigate the occurrence of abnormal patterns of HLA-ABC and HLA-DR expression in enteropathy associated T cell lymphoma and to relate such abnormalities to the Epstein Barr virus (EBV) status of the tumours. METHODS: Eleven enteropathy associated T cell lymphomas were immunostained with HC10 (HLA-ABC heavy chain) and TAL 1B5 (HLA-DR alpha chain) monoclonal antibodies and polyclonal anti-beta 2 microglobulin (beta 2m, the HLA-ABC light chain) antibodies. In situ hybridisation for EBV using EBER probes was performed on all cases. RESULTS: Tumour cells of two of 11 patients were EBER positive. One of these showed partial, and the other, complete loss of beta 2m. HLA-DR expression was undetectable in both patients. Of the remaining nine EBER negative tumours, two were HLA-ABC heavy chain negative or showed only occasional positive cells and five of nine showed partial or complete loss of the HLA-ABC light chain, beta 2m. Seven of the nine cases were either negative for HLA-DR or showed weak expression in a proportion of tumour cells. CONCLUSIONS: These data show that low or absent HLA-ABC and HLA-DR antigen expression occurs commonly in enteropathy associated T cell lymphoma. These abnormal patterns of HLA expression may be associated with escape from immune attack which, in a minority of patients, could be directed against EBV antigens.     

IFN-gamma AU-rich element removal promotes chronic IFN-gamma expression and autoimmunity in mice

We generated a mouse model with a 162 nt AU-rich element (ARE) region deletion in the 3′ untranslated region (3′UTR) of the interferon-gamma (IFN-γ) gene that results in chronic circulating serum IFN-γ levels. Mice homozygous for the ARE deletion (ARE-Del) ^−/− present both serologic and cellular abnormalities typical of patients with systemic lupus erythematosus (SLE). ARE-Del^−/− mice display increased numbers of pDCs in bone marrow and spleen. Addition of IFN-γ to Flt3-ligand (Flt3L) treated in vitro bone marrow cultures results in a 2-fold increase in pDCs with concurrent increases in IRF8 expression. Marginal zone B (MZB) cells and marginal zone macrophages (MZMs) are absent in ARE-Del^−/− mice. ARE-Del^+/− mice retain both MZB cells and MZMs and develop no or mild autoimmunity. However, low dose clodronate treatment in ARE-Del^+/− mice specifically eliminates MZMs and promotes anti-DNA antibody development and glomerulonephritis. Our findings demonstrate the consequences of a chronic IFN-γ milieu on B220⁺ cell types and in particular the impact of MZB cell loss on MZM function in autoimmunity. Furthermore, similarities between disease states in ARE-Del^−/− mice and SLE patients suggest that IFN-γ may not only be a product of SLE but may be critical for disease onset and progression.     

shRNA干扰Jurkat细胞株MDM2表达对细胞生物特性的影响

目的研究MDM2基因与白血病细胞生物学特性的关系。方法应用pSilencer4.1CMVneo构建MDM2shRNA表达载体pMDM2shRNA,采用脂质体将载体导入Jurkat细胞,利用G418筛选出稳定表达shRNA细胞后,行RTPCR和Westernblot等技术检测细胞MDM2表达水平、基础生长曲线、细胞周期变化以及UV照射后细胞生长变化。结果发现转染pMDM2shRNA的细胞MDM2mRNA及蛋白质的表达均下降>50%,细胞的生长减慢,出现一定程度的G1～S期阻滞,DNA损伤后细胞恢复较慢。结论MDM2基因表达下降后,可导致细胞增殖减慢以及降低损伤修复的能力;细胞内表达shRNA可长期敲低靶基因的表达,可作为较理想的基因功能研究细胞模型。     

枸橼醛在急性早幼粒细胞白血病细胞系K562中的作用

目的探讨枸橼醛(citral)在急性早幼粒细胞白血病细胞系K562中的作用。方法 Citral作用于体外培养K562细胞系,分别用Wright-Giemsa染色观察细胞凋亡形态;MTT法计算细胞增殖能力;DNA电泳分析DNA碎片;Annexin V-FITC/PI分析细胞凋亡变化;RT-PCR法检测凋亡通路蛋白Bcl、Bax的表达水平变化;流式细胞术分析线粒体膜电位、Caspase-3和核因子(NF)-κB水平变化。结果 Citral以时间和剂量依赖性方式诱导K562细胞凋亡,citral能够诱导线粒体膜电位减少。证实citral诱发细胞凋亡是依赖线粒体途径。结论 Citral对白血病有潜在的治疗效果,有一定的临床应用前景。     

Effect of growth factors on the expression of proto-oncogenes c-fos and c-myc in FRTL-5 cell line.

This study was performed to prove the hypothesis that oncogene expressions would have the same patterns with those of cellular growth to growth factors in FRTL-5 cells. Ribonucleic acids of FRTL-5 were extracted at 15'', 30'', 60'' and 120'' after administration of growth factors to quiescent FRTL-5, and blotted to the nitrocellulose membrane. They were hybridized with radiolabelled c-fos, c-myc and beta-actin probes. Hybridized dot blots were autoradiographed and the amount of radioactivity was measured by densitometry. Densitometric readings were used as the indices of oncogene expressions. Expressions of c-fos and c-myc were more prominent in combined administrations of TSH (10 mU/ml) and IGF-I (100 ng/ml) or IgG of Graves'' disease (Graves'' IgG; 1 mg/ml) and IGF-I than in combined administration of TSH and Graves'' IgG. IgG of primary myxedema suppressed oncogene expressions by TSH or Graves'' IgG, but not by IGF-I. From the above results, it was suggested that expressions of c-fos and c-myc to growth factors would have similar patterns with those of cell growth to growth factors in FRTL-5, and the actions of TSH and Graves'' IgG would be manifested through same signal transduction system, but IGF-I would be manifested by its own.