人正常腺上皮组织中MUC6基因的表达

应用免疫组织化学和原位杂交方法检测人正常腺上皮中MUC6基因的表达。揭示MUC6基因在正常人腺上皮组织中的分布异质性及其特点,结果显示：MUC6基因编码的核心蛋白及其mRNA主要分布于正常胃粘膜胃腺的基底部,上皮细胞无MUC6基因表达。呈细颗粒状,位于细胞核周,胃底,胃窦的表达无区别;十二指肠绒毛上皮内的表达呈弥漫性,均质状,杯状和柱状细胞的表达类似,杯状细胞的粘液滴内未测得MUC6基因产物;空肠,结肠组织中无MUC6基因的表达;胆囊上皮组织内有强阳性MUC6核心蛋白的表达。而宫颈上皮中表达较弱,实验提示;MUC6基因的表达存在异质性及器官特异性。     

人正常腺上皮组织中MUC6基因的表达

应用免疫组织化学和原位杂交方法检测人正常腺上皮中MUC6基因的表达,揭示MUC6基因在正常人腺上皮组织中的分布异质性及其特点.结果显示MUC6基因编码的核心蛋白及其mRNA主要分布于正常胃粘膜胃腺的基底部,上皮细胞无MUC6基因表达,呈细颗粒状,位于细胞核周,胃底、胃窦的表达无区别;十二指肠绒毛上皮内的表达呈弥漫性,均质状,杯状和柱状细胞的表达类似,杯状细胞的粘液滴内未测得MUC6基因产物;空肠、结肠组织中无MUC6基因的表达;胆囊上皮组织内有强阳性MUC6核心蛋白的表达,而宫颈上皮中表达较弱.实验提示MUC6基因的表达存在异质性及器官特异性.     

乳腺上皮细胞的特殊粘蛋白MUC1

乳腺上皮细胞的特殊粘蛋白ＭＵＣ１,是乳腺上皮细胞膜结合蛋白,富含唾液酸,含糖量高达５０％并能和许多植物凝集素发生特异性结合。在ＳＤＳ—ＰＡＧＥ电泳上表现为多态性,个体一般为１－２带,源自双亲两个等位基因的遗传和组织中的共同表达。ＭＵＣ１在机体怀孕中期和进入泌乳期在乳腺中水平显著提高,并受体内激素的调节。ＭＵＣ１在机体内有重要的生物学功能     

人类疱疹病毒7型体外生长特点的研究

研究了人类疱疹病毒７型（ＨＨＶ－７）南京株ＹＹ５在脐血单个核细胞（ＣＢＭＣｓ）及ＳＵＰＴ１细胞株上生长特点。观察病毒感染细胞后不同时间病变效应程度,记数死亡细胞百分率,间接免疫荧光染色记数抗原表达阳性细胞数,并用透射电镜观察病毒感染细胞超微结构变化及病毒复制不同时期的特点。结果发现：ＨＨＶ－７在ＣＢＭＣｓ及ＳＵＰＴ１上出现ＣＰＥ时间迟于ＨＨＶ－６,且ＣＰＥ程度也低于ＨＨＶ－６;ＨＨＶ－７在ＳＵＰＴ     

ICAM在着床期小鼠PBLC及EC/DC中表达的研究(英文)

细胞粘附分子（ＣＡＭ）可介导细胞间及细胞与间质之间的相互作用并传导信息,参与机体胚胎发育、免疫调节、炎症反应、组织修复及肿瘤转移等生理和病理过程。细胞间粘附分子１（ＩＣＡＭ１）是主要的ＣＡＭ分子之一,可表达于活化的细胞,内皮细胞等。人膜是母体与胚胎滋养层直接接触的特殊组织、已发现蜕膜细胞在着床过程中参与了局部免疫耐受的形成,但对着床期ＩＣＡＭ１在蜕膜细胞表达的动态研究鲜见报道。本研究采用免疫荧光、多参数流式细胞术,分别从整体和局部角度、着床过程中外周血淋巴细胞（ＰＢＬＣ）及子宫内膜／蜕膜（ＥＣ／ＤＣ）细胞ＩＣＡＭ１的不同表达特点进行了动态观察和对比性分析。结果发现,ＩＣＡＭ１在着床期ＰＢＬＣ及ＥＣ／ＤＣ中的表达均存在明显的动态变化（Ｔａｂ．１;Ｆｉｇｓ．１＆２）。ＩＣＡＭ１在ＰＢＬＣ中的表达于妊娠第一天（Ｄ１）即开始降低,Ｄ２降至最低;与此不同,ＩＣＡＭ１在ＥＣ／ＤＣ中的表达于Ｄ２开始降低,Ｄ４降至最低,Ｄ５开始恢复,但尚未恢复到对照水平。结果表明,ＩＣＡＭ１在蜕膜局部的表达调节方式不同于外周血;ＩＣＡＭ１表达阳性的外周血,淋巴细胞和蜕膜细胞均代表着活化的功能性细胞,这些细胞表面ＩＣＡＭ     

EB病毒潜伏膜蛋白1基因对上皮细胞增殖的影响

为了研究ＥＢ病毒潜伏膜蛋白１（ＬＭＰ１）基因对上皮细胞增殖的影响,探索ＬＭＰ１在上皮细胞肿瘤发生中所起的作用。用ＬＭＰ１基因真核表达质粒转染人胚肾上皮细胞,检测了转染细胞中ＬＭＰ１的表达,观察细胞在软琼脂中的集落形成能力、ＭＴＴ吸收能力以及ＰＣＮＡ的表达情况。结果显示,被ＬＭＰ１基因转染的细胞生长旺盛,能在软琼脂中形成多个集落,ＭＴＴ吸收能力增强,ＰＣＮＡ的表达水平增高。因此认为ＬＭＰ１基因能明显改变上皮细胞的生物学行为,促进细胞的生长、增殖和转化,使转染的上皮细胞获得肿瘤细胞的生长特征     

EB病毒转化Hu—TLC—SCID小鼠脾细胞的实验研究

利用ＥＢ病毒转化可产生较高水平人ＩｇＧ和特异性抗２型登革病毒人抗体的Ｈｕ－ＴＬＣ－ＳＣＩＤ小鼠脾细胞,通过免疫组化、免疫荧光和ＰＣＲ法检测转化细胞的人Ｂ细胞表面标志、ＥＢ病毒抗原和ＥＢ病毒基因。结果表明,被团体的Ｈｕ－ＴＬＣ－ＳＣＩＤ小鼠脾细胞能继续产生抗２型登革病毒的特异性人抗体,并具有人Ｂ细胞的ＣＤ２０＾＋、ＳｍＩｇＧ标志及ＥＢ病毒潜伏膜蛋白－１（ＬＭＰ－１）基因,可表达ＬＭＰ－１和ＥＢ病毒核     

EB病毒转化Hu－TLC－SCID小鼠脾细胞的实验研究

利用ＥＢ病毒转化可产生较高水平人ＩｇＧ和特异性抗２型登革病毒人抗体的Ｈｕ－ＴＬＣ－ＳＣＩＤ小鼠脾细胞,通过免疫组化、免疫荧光和ＰＣＲ法检测转化细胞的人Ｂ细胞表面标志、ＥＢ病毒抗原和ＥＢ病毒基因。结果表明,被转化的Ｈｕ－ＴＬＣ－ＳＣＩＤ小鼠脾细胞能继续产生抗２型登革病毒的特异性人抗体,并具有人Ｂ细胞的、ＳｍＩｇＧ标志及ＥＢ病毒潜伏膜蛋白－１（ＬＭＰ－１）基因,可表达ＬＭＰ－１和ＥＢ病毒核抗原（ＥＢＮＡ）。     

BRCA1基因结构及其在乳腺癌中的表达

利用ＭＰＣＲ技术从乳腺组织分离到抑癌基因ＢＲＣＡ１的ｃＤＮＡ片段,将此９１４ｂｐ的片段克隆进质粒ｐＵＣ１１８,并经全序列测定证实。序列分析表明,ＢＲＣＡＩｃＤＮＡ编码的肽链ＮＮ２－末端有一锌指结构,抑癌基因ＢＲＣＡＩ的产物可能是ＤＮＡ结合蛋白,ｃＤＮＡ序列存在两个变异位点：一个是第４０９位的Ｃ→Ａ（Ａｓｐ→Ｇｌｕ）：另一个是第８７９位的Ａ→Ｔ（ＭＩａ同义突变）。以该片段为探针,检测６例乳腺癌组织中ＢＲＣＡＩｍＩＭＡ表达,一例表达明显下降,一例没检测到表达的ｍＩＭＡ产物,说明一些乳腺癌组织的ＢＲＣＡ１基因转录水平降低     

EB病毒LMP1在鼻咽癌细胞中通过NFκB促进Igκ表达

利用已建立的受四环素调控ＬＭＰ１表达的鼻咽癌细胞系,用受ＣＭＶ启动子调控的ＮＦκＢ报道基因质粒ｐＮＦκＢ－ｌｕｃ的荧光素酶表达分析ＮＦκＢ的活性,并以核蛋白的Ｗｅｓｔｅｒｎ印迹方法观察ＬＭＰ１表达前后核内ＮＦκＢ组分ｐ６５量的改变,用全蛋白Ｗｅｓｔｅｒｎ印迹分析Ｉｇκ蛋白质的表达等方法,探讨在鼻咽癌中,ＥＢ病毒ＬＭＰ１蛋白是否通过核转录因子ＮＦκＢ促进免疫球蛋白κ轻链（Ｉｇκ）基因的表达。结果显示     

Variation of human mucin gene expression in gastric cancer cell lines and gastric mucous cell primary cultures

Human gastric mucous cells - gastric cancer cell lines mucin gene expression - TNFalpha - RT-PCR immunocytochemistry Little is known on the expression pattern of mucin genes in human gastric cancer cell lines in relation to mucin expression in normal gastric epithelial cells. Thus, the aim of this study was to compare gastric cancer cell lines and non-transformed epithelial cells in their expression of the different mucin genes, in order to use these cells as models for physiological MUC expression in human stomach. Human gastric mucous cell primary cultures which were obtained from surgical specimen by collagenase/pronase treatment and a panel of six human gastric cancer cells were screened for mRNA expression of the mucin genes MUC1, MUC2, MUC5AC, MUC5B, and MUC6. Mucin gene expression was analyzed by semi-quantitative RT-PCR, and by Western blotting and immunocytochemistry. Primary cultured human gastric mucous cells retained the stomach-specific pattern of mRNA expression found in gastric mucosal biopsies (MUC1, MUC5AC, MUC6), whereas any gastric cancer cell line exhibited an aberrant mucin gene expression. Mucin gene expression showed large variations in levels and patterns from cell line to cell line, but MUC2 was aberrantly expressed in all cancer cells. Immunocytochemistry confirmed aberrant MUC2 protein expression in cancer cells. The expression of the secretory mucin genes MUC2 and MUC5AC varied in relation to the length of cultivation of the cancer cell lines. Treatment of the gastric cancer cells with TNFalpha resulted in an enhanced mRNA expression of MUC1, MUC2, and MUC5AC (2-fold increase within 3 hours; p <0.05). In contrast, immunocytochemistry disclosed a decrease in MUC2 and MUC5AC staining intensity. Our results indicate that primary cultured human gastric mucous cells provide a physiological in vitro system for investigations of gastric mucin gene regulation. In gastric cancer cells marked changes in the mucin gene expression pattern are found with coexpression of non-gastric type mucins. Gastric mucin gene expression may be regulated by proinflammatory cytokines which could have implications in gastritis.     

IL-4 induces mucin gene expression and goblet cell metaplasia in vitro and in vivo

Goblet cell metaplasia and mucus hypersecretion are important features in the pathogenesis of asthma. The cytokine IL-4 has been shown to play a role in animal models of asthma, where it induces Th2 lymphocyte differentiation and B lymphocyte IgE class switch. IL-4 has also been implicated in the differentiation of goblet cells via effects on lymphocytes and eosinophils. In this study we hypothesized that IL-4 induces airway epithelial cell mucin gene expression and mucous glycoconjugate production by direct action on these cells. In vitro, cultured airway epithelial cells (NCI-H292) expressed IL-4R constitutively, and IL-4 (10 ng/ml) induced MUC2 gene expression and mucous glycoconjugate production. In vivo, mouse airway epithelial cells expressed IL-4R constitutively, and IL-4 (250 ng) increased MUC5 gene expression and Alcian blue/periodic acid-Schiff-positive staining at 24 h; IL-4 did not increase inflammatory cell numbers in airway tissue or in bronchoalveolar lavage. TNF-alpha and IL-1beta levels in bronchoalveolar lavage were not increased in response to IL-4 instillation. These results indicate that airway epithelial cells express IL-4R constitutively and that IL-4 directly induces the differentiation of epithelium into mucous glycoconjugate-containing goblet cells.     

Interleukin-1 beta and tumor necrosis factor-alpha induce MUC5AC overexpression through a mechanism involving ERK/p38 mitogen-activated protein kinases-MSK1-CREB activation in human airway epithelial cells

Mucin hypersecretion is commonly observed in many inflammatory diseases of the respiratory tract. MUC5AC is generally recognized to be a major airway mucin because MUC5AC is highly expressed in the goblet cells of human airway epithelium. Moreover, it is regulated by various inflammatory cytokines. However, the mechanisms by which the interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha induce MUC5AC gene expression in normal nasal epithelial cells, and the signal molecules involved, especially in the downstream signaling of mitogen-activated protein (MAP) kinases, remain unclear. Here we show that pharmacologic or genetic inhibition of either ERK or p38 MAP kinase pathway abolished IL-1beta- and TNF-alpha-induced MUC5AC gene expression in normal human nasal epithelial cells. Our results also indicate that the activation of mitogen- and stress-activated protein kinase 1 (MSK1) and cAMP-response element-binding protein and cAMP-response element signaling cascades via ERK and p38 MAP kinases are crucial aspects of the intracellular mechanisms that mediate MUC5AC gene expression. Taken together, these studies give additional insights into the molecular mechanism of IL-1beta- and TNF-alpha-induced MUC5AC gene expression and enhance our understanding on mucin hypersecretion during inflammation.     

The Human Endometrium Expresses the Glycoprotein Mucin-1 and Shows Positive Correlation for Thomsen-Friedenreich Epitope Expression and Galectin-1 Binding

Mucin 1 (MUC1) is a glycoprotein in human endometrium and is abundant at the luminal epithelial surface in the receptive phase. It has a highly glycosylated ecto-domain that contains keratan sulfate chains, that disappears at the time of implantation. In addition, the glycoforms on MUC1 differ in fertile and infertile women. Therefore the aims of this study were investigations on glycosylation of MUC1 with the Thomsen-Friedenreich (TF) epitope on normal human endometrium throughout the menstrual cycle and binding of galectin-1 on the TF epitope in the endometrium and the expression of galectin-1 on the human oocyte. Human endometrial tissue was obtained from 54 premenopausal patients and was immunohistochemically analyzed with monoclonal antibodies against MUC1, TF epitope, galectin-1, and biotinylated galectin-1. In addition, human oocytes were analyzed for TF, galectin-1 expression, and galectin-1 binding. We identified a significant upregulation of MUC1 and TF epitope and, in addition, galectin-1 binding in glandular epithelium and epithelial apical surface tissue from proliferative to secretory phase. With double staining experiments, we identified a coexpression of TF and MUC1 in the early secretory phase and galectin-1 binding to TF during the same period of time. In addition we identified TF epitope and galectin-1 expression plus binding on the human oocyte and irregularly fertilized oocytes. Upregulation of TF epitope on the glandular epithelium and epithelial apical surface tissue in the secretory phase and binding of galectin-1 at the same time show the possibility of galectin-1–mediated trophectoderm binding to the endometrium within the window of implantation. (J Histochem Cytochem 57:871–881, 2009)     

Delta-like 1 expression promotes goblet cell differentiation in Notch-inactivated human colonic epithelial cells

Notch signaling has previously been implicated in the regulation of the cell fate of intestinal epithelial cells. However, the expression and function of Notch ligands in the human intestine remain largely unknown. In the present study, we showed that Notch ligands Delta-like 1 (Dll1) and Delta-like 4 (Dll4) are expressed in a goblet cell-specific manner in human colonic tissue. Additionally, we found that Dll1 and Dll4 expression was regulated in-parallel with Atoh1 and MUC2, which are both under the control of the Notch-Hes1 signaling pathway. Because knockdown of Dll1 expression completely abrogated the acquisition of the goblet cell phenotype in Notch-inactivated colonic epithelial cells, we postulate that Dll1 might function as a cis-acting regulatory element that induces undifferentiated cells to become goblet cells. Our results suggest a link between Dll1 expression and human goblet cell differentiation that might be mediated by a function that is distinct from its role as a Notch receptor ligand.     

Differential glycosylation of MUC1 in tumors and transfected epithelial and lymphoblastoid cell lines

The membrane-bound mucin-like protein MUC1 with a specified number of tandem repeats has been expressed by transfection of the cDNAs in both the epithelial cell lines MDCK and LLC-PK1, and human lymphoblastoid cell lines T2 and C1R. The structure and glycosylation states of the MUC1 in these four lines were compared with that of the endogenous MUC1 found in the human pancreatic (HPAF) and breast (BT-20) tumor cell lines using flow cytometry and Western blot analysis with anti-MUC1 antibodies, which are either sensitive or insensitive to the glycosylation state of the tandem repeat, and pretreatment of cells with phenyl--galactosaminide, an inhibitor of mucin sialylation. A similar analysis of MUC1 expression in transfected normal and O-glycosylation defective CHO cells reveals that the addition of galactose to the core oligosaccharide structure is apparently responsible for the anomalous difference in Mr between the mature and propeptide forms of the MUC1. Both the tumor cells and the transfected lymphoblastoid cells consistently express significant steady state levels of both the heavily glycosylated mature forms and the poorly glycosylated propeptide forms of the MUC1, whereas MUC1 is found predominantly as the mature extensively glycosylated species in the transfected epithelial cells. Immunofluoresence microscopy of cross sections of the polarized epithelial cells grown on culture filter inserts reveals that the MUC1 is clearly present at the apical surface of the cells, consistent with its expression in normal tissues. Thus, the successful expression of the MUC1 by transfection of either lymphoblastoid cells or epithelial cells yields model systems both for studying the natural structure/function relationships of the protein domains within the MUC1 molecule and for further elucidating the previously reported MHC-independent T-cell recognition of the MUC1.