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Regulation of tissue factor gene expression in human endometrium by transcription factors Sp1 and Sp3

MOLECULAR ENDOCRINOLOGY 2000年第3期14卷 393-400页

作者： Krikun, G Schatz, F Mackman, N Guller, S Demopoulos, R Lockwood, CJ NYU Sch Med Dept Obstet & Gynecol New York NY 10016 USA NYU Sch Med Dept Biochem New York NY 10016 USA NYU Sch Med Dept Pathol New York NY 10016 USA Scripps Res Inst Dept Immunol La Jolla CA 92037 USA Scripps Res Inst Dept Vasc Biol La Jolla CA 92037 USA

Prior studies indicate that tissue factor (TF), the primary cellular initiator of hemostasis, is persistently up-regulated in human endometrial stromal cells (HESCs) undergoing progestin-induced decidualization in vivo and in vitro. The mechanism underlying progestin enhancement of TF mRNA and protein levels in these cells involves transcriptional activation of the TF gene. Transient transfections of HESCs with the truncated TF promoters driving the luciferase reporter gene have demonstrated that the region spanning -111 to +14 bp retained differential progestin-enhancing effects. We now demonstrate that RU486 displays inhibitory effects on the progestin-induced TF promoter activity, confirming the involvement of the progesterone receptor. Since the TF minimal promoter (pTF 111 spanning -111 to +14 bp) contains three overlapping Sp1 and three Egr-1 sites, the present study determined whether Sp1 and/or Egr-1 were required for progestin-regulated TF expression. The results indicate that the three Sp1 sites are primarily responsible for both the constitutive and progestational activity of the pTF 111 promoter, whereas the Egr-1 sites have only a minor involvement in both activities. Overexpression of the Sp1 protein resulted in greater than a 6-fold induction in TF promoter activity. In contrast, no enhancement was observed when the Sp3 protein was overexpressed. The concomitant overexpression of Sp1 and Sp3 demonstrated that Sp3 completely blocked the induction of TF promoter activity by Sp1. Moreover, the addition of 10 nM mithramycin, a concentration that inhibits Sp1 binding to target DNA, blocked the progestational induction of TF mRNA expression. Immunohistochemical studies demonstrated increased Sp1 levels in perivascular stromal cells in secretory phase compared with proliferative phase endometria. In contrast, Sp3 expression was greatly decreased in stromal cells of secretory, compared with proliferative phase tissues. The levels of Egr-1 were low in both pro

关键词：碱基序列细胞培养的/药物作用 DNA结合蛋白质类/生理学早期生长反应蛋白质1 子宫内膜/药物作用子宫内膜/代谢子宫内膜/分泌雌二醇/药理学基因表达调控/药物作用激素拮抗药/药理学即早蛋白质类甲羟孕酮17-乙酸酯/药理学月经周期米非司酮/药理学分子序列数据孕激素类/药理学启动区(遗传学) RNA 信使/生物合成重组融合蛋白质类/生理学 Sp1转录因子/生理学 Sp3转录因子间质细胞/药物作用凝血致活酶/生物合成凝血致活酶/遗传学转录因子/生理学转录遗传/药物作用女(雌)性人类

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Effects of matrix components on aromatase activity in breast stromal cells in culture

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JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY 1999年第4-6期70卷 249-256页

作者： Quinn, AL Burak, WE Brueggemeier, RW Ohio State Univ Ohio State Biochem Program Columbus OH 43210 USA Ohio State Univ Coll Med Dept Surg Columbus OH 43210 USA Ohio State Univ Ctr Comprehens Canc Columbus OH 43210 USA Ohio State Univ Coll Pharm Div Med Chem & Pharmacognosy Columbus OH 43210 USA

Local estradiol production within breast tissue is maintained by the aromatase cytochrome P450(arom) complex, which has been localized primarily to the stromal component of tumors but also has been detected in the breast epithelial cells. Paracrine interactions between stromal and epithelial components of the breast are critical to the sustained growth and progression of breast tumors. Maintenance of the differentiated state, including hormone and growth factor responsiveness, requires extracellular matrix proteins as substrata for cells. This research has focused on developing a cell culture system that more closely mimics in vivo interactions in order to dissect actual paracrine signaling between these two cell types. Human fibroblasts were isolated from breast tissue and were maintained in a cell culture system grown on plastic support or on a collagen I support matrix. The collagen I matrix model supports cell maintenance and subsequent differentiation on collagen rather than maximal proliferation, therefore allowing for a more accurate environment for the study of hormonal control and cellular communication. Initial experiments compared aromatase activity of patient fibroblasts grown on plastic versus collagen I using the tritiated water release method. Constitutive aromatase activity was found to be lower when cells were grown on a collagen gel for 4-7 days (7.7 fold lower) using DMEM/F12 containing 10% dextran coated charcoal stripped serum. However, fibroblasts grown on collagen I appeared to be significantly more responsive to stimulation by 100 nM dexamethasone (plastic: 6.0 fold induction, collagen: 33.2 fold induction) when pretreated for 12 h prior to measurement of aromatase activity. In an effort to examine paracrine interactions between the stromal and epithelial cells in breast tissue, experiments using conditioned media from fibroblast cultures were performed. Testosterone administration to fibroblasts results in the production of estradiol into th

关键词：腺癌/酶学芳香酶/代谢乳房/细胞学乳房/药物作用乳房/酶学乳腺肿瘤/酶学细胞分裂细胞培养的胶原/药理学地塞米松/药理学上皮细胞/生理学细胞外基质基因表达调控/药物作用 RNA 信使/遗传学间质细胞/药物作用间质细胞/酶学睾酮/药理学胸苷/代谢肿瘤细胞培养的波形蛋白/遗传学女(雌)性人类

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Interleukin-7 stimulates osteoclast formation by up-regulating the T-cell production of soluble osteoclastogenic cytokines

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BLOOD 2000年第5期96卷 1873-1878页

作者： Weitzmann, MN Cenci, S Rifas, L Brown, C Pacifici, R Barnes Jewish Hosp Div Bone & Mineral Dis St Louis MO 63110 USA Washington Univ Sch Med Div Bone & Mineral Dis St Louis MO USA Univ Calgary Div Hematol Calgary AB Canada

In unstimulated conditions osteoclast renewal occurs as a result of the stromal cell production of the key osteoclastogenic factors, receptor activator of NFkB ligand (RANKL) and macrophage colony-stimulating factor (M-CSF), Inflammation is known to cause increased osteoclastogenesis;however, the mechanisms responsible for this phenomenon are poorly understood. We now show that interleukin-1 (II-1) and tumor necrosis factor alpha (TNF alpha), cytokines typically produced in inflammatory conditions, increase the stromal cell production of IL-7, This factor, in turn, up-regulates production of osteoclastogenic cytokines by T cells leading to stimulation of osteoclast (OC) formation. Although T cells were found to produce soluble forms of both RANKL and M-CSF, saturating concentrations of osteoprotegerin failed to inhibit approximately 40% of the OC formation, suggesting that IL-7 acts via both RANKL-dependent and RANKL-independent pathways, Despite the identification of T-cell-secreted M-CSF, this cytokine was not essential for either RANKL-dependent or -independent OC formation, suggesting that T cells secrete other cytokines capable of substituting for M-CSF action, On the basis of our data, we propose a novel mechanism for inflammatory bone loss in which induction of IL-7 from stromal cells by IL-l and TNF alpha leads to the production of soluble osteoclastogenic cytokines by T cells. Thus, the mechanism by which IL-7 causes bone resorption involves the activation of T cells and the T-cell-dependent augmentation of osteoclastogenesis, (C) 2000 by The American Society of Hematology.

关键词：酸性磷酸酶/药物作用酸性磷酸酶/代谢载体蛋白质类/药物作用载体蛋白质类/遗传学载体蛋白质类/代谢细胞培养的细胞因子类/药物作用细胞因子类/代谢剂量效应关系药物造血干细胞/细胞学造血干细胞/药物作用白细胞介素1/药理学白细胞介素7/代谢白细胞介素7/药理学同工酶类/药物作用同工酶类/代谢膜糖蛋白类/药物作用膜糖蛋白类/遗传学膜糖蛋白类/代谢成骨细胞/细胞学成骨细胞/药物作用成骨细胞/代谢 RANK配体 RNA 信使/药物作用 RNA 信使/遗传学 RNA 信使/代谢核因子κB受体活化因子重组蛋白质类/代谢重组蛋白质类/药理学间质细胞/细胞学间质细胞/药物作用间质细胞/代谢 T淋巴细胞/细胞学 T淋巴细胞/药物作用 T淋巴细胞/代谢肿瘤坏死因子α/药理学增量调节人类

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Enhancement of osteogenesis in vitro and in vivo by a novel osteoblast differentiation promoting compound, TAK-778

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 1999年第3期290卷 1054-1064页

作者： Notoya, K Nagai, H Oda, T Gotoh, M Hoshino, T Muranishi, H Taketomi, S Sohda, T Makino, H Takeda Chem Ind Ltd Pharmaceut Res Labs 4 Div Pharmaceut Res Yodogawa Ku Osaka 5328686 Japan Takeda Chem Ind Ltd Drug Delivery Syst Res Labs Div Pharmaceut Res Osaka 5328686 Japan Takeda Chem Ind Ltd Pharmacol Labs Div Pharmaceut Res Osaka 5328686 Japan Takeda Chem Ind Ltd Drug Safety Res Labs Pharmaceut Dev Div Osaka 532 Japan

TAK-778 [(2R,4S)-(-)-N-(4-diethoxyphosphorylmethylphenyl)- 1,2,4,5-tetrahydro-4-methyl-7,8-methylenedioxy-5-oxo-3-benzothiepin -2-carboxyamide;mw 505.53], a novel osteoblast differentiation promoting compound, was characterized in vitro and in vivo models. TAK-778 at doses of 10(-6) M and higher promoted potently bone-like nodule formation in the presence of dexamethasone in rat bone marrow stromal cell culture. This was accompanied by increases in cellular alkaline phosphatase activity, soluble collagen release, and osteocalcin secretion. Under the culture conditions, TAK-778 also stimulated the secretion of transforming growth factor-p and insulin-like growth factor-I, indicating that TAK-778 may exert regulatory effects on osteoblast differentiation via autocrine/paracrine mechanisms. Furthermore, the in vivo osteogenic potential of TAK-778 was studied in bony defect and osteotomy animal models, using sustained release microcapsules consisted of a biodegradable polymer, poly (dl-lactic/glycolic) acid (PLGA). Single local injection of TAK-778/PLGA-microcapsules (PLGA-MC) (0.2-5 mg/site) to rat skull defects resulted in a dose-dependent increase in new bone area within the defects after 4 weeks. When the pellet containing TAK-778/PLGA-MC (4 mg/pellet) was packed into place to fill the tibial segmental defect in rabbit, this pellet induced osseous union within 2 months, whereas the placebo pellet did not. In addition, single local application of TAK-778/PLGA-MC (10 mg/site) to rabbit tibial osteotomy site enhanced callus formation accompanied by an increase in breaking force after 30 days. These results reveal for the first time that a nonendogenous chemical compound promotes potently osteogenesis in vitro and enhances new bone formation during skeletal regeneration and bone repair in vivo and should be useful for the stimulation of fracture healing.

关键词：苯硫|艹卓|类/投药和剂量苯硫|艹卓|类/药理学生物相容性材料/投药和剂量骨髓细胞/细胞学骨髓细胞/药物作用骨再生/药物作用细胞分化/药物作用细胞培养的乳酸/投药和剂量小鼠近交C3H 成骨细胞/细胞学成骨细胞/药物作用成骨细胞/酶学骨生成/药物作用聚乙醇酸/投药和剂量聚合物/投药和剂量大鼠 Sprague-Dawley 颅骨/药物作用颅骨/损伤间质细胞/细胞学间质细胞/药物作用胫骨骨折/药物疗法动物男(雄)性小鼠兔大鼠

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Interleukin-1 (IL-1) inhibits growth of cytomegalovirus in human marrow stromal cells: Inhibition is reversed upon removal of IL-1

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BLOOD 1999年第2期94卷 572-578页

作者： Iwata, M Vieira, J Byrne, M Horton, H Torok-Storb, B Fred Hutchinson Canc Res Ctr Div Clin Res Seattle WA 98109 USA Genet Inst Cambridge MA 02140 USA

A Toledo strain cytomegalovirus (CMV) containing the gene for green fluorescent protein (GFP) under the control of elongation factor-1 promoter was used to study infection of human marrow stromal cells. Two stromal cell lines were used: HS-5, which secretes copious amounts of known cytokines and interleukins;and HS-27a, which does not secrete these activities. CMV growth and spread was monitored by counting green plaques and quantitating GFP intensity. Initial studies indicated that, whereas HS-5 and 27a have similar susceptibilities to infection, as evidenced by the same number of GFP(+) cells at day 2, HS-5 appears more resistant to growth and spread of CMV. Furthermore, conditioned media from HS-5 (HS-5 CM) inhibited CMV plaque formation in HS-27a, suggesting that factors secreted by HS-5 are responsible for limiting CMV growth. Neutralizing antibodies against interleukin-1 alpha (IL-1 alpha) and IL-1 beta completely blocked the ability of HS-5 CM to limit viral growth, suggesting that IL-1, which is known to be present in HS-5 CM, is responsible for this effect. When exogenous IL-1 beta was added to CMV-infected HS-27a, both the number of plaques and the intensity of GFP was significantly reduced in IL-1-treated HS 27a compared with untreated HS-27a (the number of plaques by day 18 was 20 +/- 3 v 151 +/- 12/well, respectively;GFP intensity was 535 +/- 165 v 6,516 +/- 652/well, respectively, in 4 separate experiments). At day 21, when IL-1 beta-treated, CMV-infected cultures were passaged and then cultured in the absence of IL-1 beta, CMV growth progressed with the kinetics of the original untreated culture, indicating that the IL-1 beta effect is reversible. Because HS-27a expresses the type I IL-l receptor, we speculate that the antiviral effects are mediated through IL-1-induced changes in cellular gene expression. DNA chip analysis of mRNA from IL-1 beta-treated and nontreated HS-27a cells has identified some candidate molecules, (C) 1999 by The American Soci

关键词：骨髓细胞/药物作用骨髓细胞/病毒学细胞培养的巨细胞病毒/药物作用巨细胞病毒/生长和发育巨细胞病毒/生理学基因报告绿色荧光蛋白质类白细胞介素1/药理学发光蛋白质类/生物合成发光蛋白质类/遗传学肽延伸因子1 肽延伸因子/遗传学启动区(遗传学) 间质细胞/药物作用间质细胞/病毒学病毒复制/药物作用人类

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Adenomyosis - A result of disordered stromal differentiation

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AMERICAN JOURNAL OF PATHOLOGY 2001年第2期159卷 623-630页

作者： Parrott, E Butterworth, M Green, A White, INH Greaves, P MRC Toxicol Unit Leicester LE1 9HN Leics England

Adenomyosis is a fairly frequent disorder in adult women characterized by the haphazard location of endometrial glands and stroma deep within the myometrium of the uterus. This study compared the effects on uterine development of the selective estrogen receptor modulators, tamoxifen, toremifene, and raloxifene with estradiol when given orally to female mice on days 2 to 5 after birth. Uterine adenomyosis was found in all (14 of 14) mice dosed with tamoxifen and most mice (12 of 14) treated with toremifene, but in none of the vehicle-dosed controls, in only one animal treated with raloxifene at 42 and 90 days after dosing and in none of the mice treated with estradiol at 42 days. At 6 days, the uterus in the groups that developed a high incidence of adenomyosis showed histological evidence of disturbed differentiation of the myometrium. Gene-expression XY-scatterplots using Clontech mouse 1.2 Atlas mouse cDNA expression arrays analyzing total uterine RNA showed nerve growth factor-alpha, preadipocyte factor-1, and insulinlike growth factor-2 were key genes differentially modified by tamoxifen or toremifene treatment, relative to the controls. As these genes may play an important role in regulating differentiation and development of the myometrium, these data suggest that adenomyosis may be caused primarily by defects in the formation of the myometrium.

关键词：投药口服衰老动物新生体重/药物作用细胞分化/药物作用子宫内膜异位症/病理学雌二醇/投药和剂量雌二醇/药理学基因表达调控/药物作用生长抑制物/遗传学胰岛素样生长因子Ⅱ/遗传学胞间信号肽类和蛋白质类膜蛋白质类/遗传学子宫肌层/细胞学子宫肌层/药物作用子宫肌层/病理学神经生长因子/遗传学寡核苷酸序列分析器官测量/药物作用雷洛昔芬/药理学阻遏蛋白质类/遗传学选择性雌激素受体调节剂/药理学间质细胞/细胞学间质细胞/药物作用间质细胞/病理学他莫昔芬/药理学托瑞米芬/药理学子宫/细胞学子宫/药物作用子宫/生长和发育动物女(雌)性小鼠

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Characterization of human intestinal stromal cell lines: Response to cytokines and interactions with epithelial cells

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EXPERIMENTAL CELL RESEARCH 1999年第2期248卷 391-406页

作者： Fritsch, C Orian-Rousseaul, V Lefebvre, O Simon-Assmann, P Reimund, JM Duclos, B Kedinger, M INSERM U381 F-67200 Strasbourg France Univ Hosp Hautepierre Dept Gastroenterol Strasbourg France

The maintenance of the physiological homeostasis of the gut mucosa characterized by continuous proliferation and differentiation processes results from epithelial-mesenchymal cell cross-talk. To set out stable and homogeneous models for the study of the (dys)regulation of various morphofunctional aspects, we established and characterized three clonal cell lines (C9, C11, and C20) derived from human duodenal mucosal connective tissue. We defined the expression of (i) cytoskeletal proteins;(ii) basement membrane molecules (laminins, collagen IV, nidogen) which have been shown formerly to be deposited at the epithelial/mesenchymal interface in situ by the mesenchymal compartment;and (iii) soluble factors, HGF, and TGF beta 1. The three cell lines display common but also specific proliferative responses to cytokines (IL1 beta, IL2, IL8, TNF alpha, IFN gamma, TGF beta 1, and HGF). When cocultured with embryonic intestinal endoderms or with human colonic Caco2 or HT29 cancer cells, C9 versus C11 and C20 cell lines induced limited versus extensive growth of the associated epithelial cells. In addition C20 cells allowed spreading of HT29 cells with the formation of a basement membrane at the heterologous interface. Morphogenesis obtained by intracoelomic grafts of associations comprising the mesenchymal cell lines and intestinal endoderms was also different among those composed of C9 cells or of C11 or C20 cells. In conclusion, these data indicate that the mucosal connective tissue is heterogeneous and comprises several phenotypically different mesenchyme-derived cells whose equilibrium may be important in the gut homeostasis. These cells can now be used to define tissue-specific factors which may be involved in the physiopathology of the intestinal epithelium. (C) 1999 Academic Press.

关键词：基膜/化学 Caco-2细胞细胞间通讯细胞系共同培养技术胶原/分离和提纯细胞因子类/药理学十二指肠/细胞学十二指肠/药物作用十二指肠/生理学内胚层/移植上皮细胞/细胞学上皮细胞/药物作用上皮细胞/生理学 HT29细胞内环境稳定肠黏膜/细胞学肠黏膜/生理学层黏连蛋白/分离和提纯中胚层/细胞学中胚层/药物作用中胚层/生理学中胚层/移植形态发生间质细胞/细胞学间质细胞/药物作用间质细胞/生理学动物鸡胚人类大鼠

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Interleukin-1α is a paracrine inducer of FGF7, a key epithelial growth factor in benign prostatic hyperplasia

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AMERICAN JOURNAL OF PATHOLOGY 2000年第1期157卷 249-255页

作者： Giri, D Ittmann, M Houston Dept Vet Affairs Med Ctr Res Serv 151 Houston TX 77030 USA Baylor Coll Med Dept Pathol Houston TX 77030 USA

Benign prostatic hyperplasia (BPN) is an extremely common disease of older men In which there is benign overgrowth of the prostatic transition zone, leading to obstruction of urine outflow, FGF7, a potent growth factor for prostatic epithelial cells, is increased by threefold in BPH and is correlated with increased epithelial proliferation in this condition. Immunohistochemistry of normal and hyperplastic prostate revealed that FGF7-expressing fibroblastic cells were present in higher numbers near the epithelial acini, implying that epithelial cells may express a factor that induces FGF7 expression by stromal cells. Conditioned medium (CM) from primary cultures of prostatic epithelial cells was capable of inducing a two- to sixfold increase in time expression of FGF7 by primary stromal cultures. Blocking experiments with neutralizing anti-interleukin-1 alpha(I1-1 alpha) antibodies and IL-1Ra, an I1-1 alpha receptor antagonist, show that this inducing activity was due to the presence of I1-1 alpha in the epithelial CM. Analysis of normal prostatic peripheral zone and BPH tissue by enzyme-linked immunoabsorption assay reveal that I1-1 alpha is present at increased levels in hyperplastic prostate and that levels of I1-1 alpha correlate strongly with tissue FGF7 concentration In BPH, Therefore I1-1 alpha is produced by prostatic epithelial cells and can induce FGF7, a potent epithelial growth factor, which can in turn lead to further epithelial growth and increased I1-1 alpha secretion, thus establishing a double paracrine loop that is functionally equivalent to an autocrine growth loop. This double paracrine loop may play a key role in the abnormal proliferation of the transition zone, which is critical to the pathogenesis of BPH.

关键词：细胞培养的培养基条件性/药理学剂量效应关系药物上皮细胞/细胞学上皮细胞/药物作用上皮细胞/代谢成纤维细胞生长因子10 成纤维细胞生长因子7 成纤维细胞生长因子生长物质/代谢白细胞介素1/代谢白细胞介素1/药理学旁分泌细胞细胞间通讯前列腺增生/代谢前列腺增生/病理学间质细胞/细胞学间质细胞/药物作用间质细胞/代谢时间因素人类男(雄)性

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CCAAT/enhancer-binding proteins are mediators in the protein kinase A-dependent activation of the decidual prolactin promoter

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JOURNAL OF BIOLOGICAL CHEMISTRY 1999年第35期274卷 24808-24818页

作者： Pohnke, Y Kempf, R Gellersen, B Univ Hamburg IHF Inst Hormone & Fertil Res D-252529 Hamburg Germany

In the course of decidualization, human endometrial stromal cells (ESC) activate the alternative upstream promoter of the decidual prolactin (dPRL) gene. The dPRL promoter is induced by the protein kinase A pathway in a delayed fashion via the region -332/-270 which contains two overlapping consensus binding sequences, B and D, for CCAAT/enhancer-binding proteins (C/EBP). Here we show that sites B and D both bind C/EBP beta and -delta from ESC nuclear extracts. When decidualization of cultured ESC was induced by treatment with 8-Br-cAMP, complex formation on sites B and D was enhanced. Western blot analysis revealed an elevation of both C/EBP beta isoforms, liver-enriched activator protein and liver-enriched inhibitory protein, with a delayed onset between 8 and 24 h of cAMP treatment, while C/EBP delta expression remained unaffected. Cyclic AMP-mediated activation of dPRL promoter construct dPRL-332/luc3 was abrogated by mutation of sites B and D at -310/-285. An expression vector for liver-enriched activator protein potently induced transcription of dPRL-332/luc3 and further enhanced cAMP-mediated induction, while liver-enriched inhibitory protein expression vector abolished the cAMP response, implying that C/EBPs serve as mediators in the delayed cAMP signal transduction to the dPRL promoter. The ratio between activating and repressing isoforms is likely to dictate the transcriptional output.

关键词： 8-溴环腺苷一磷酸/药理学碱基序列 CCAAT增强子结合蛋白β CCAAT增强子结合蛋白类 COS细胞环AMP依赖性蛋白激酶类/代谢 DNA结合蛋白质类/代谢蜕膜增强子元件遗传学基因表达调控基因报告分子序列数据核蛋白质类/代谢催乳素/遗传学启动区遗传蛋白质亚型/代谢阻遏蛋白质类/代谢间质细胞/药物作用间质细胞/代谢转录因子/遗传学动物女(雌)性人类

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Regulated on activation, normal T-cell-expressed and -secreted mRNA expression in normal endometrium and endometriotic implants -: Assessment of autocrine/paracrine regulation by in situ hybridization

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AMERICAN JOURNAL OF PATHOLOGY 2001年第6期158卷 1949-1954页

作者： Hornung, D Klingel, K Dohrn, K Kandolf, R Wallwiener, D Taylor, RN Univ Calif San Francisco Ctr Reprod Sci San Francisco CA 94143 USA Univ Tubingen Dept Obstet & Gynecol D-7400 Tubingen Germany Univ Tubingen Dept Mol Pathol D-7400 Tubingen Germany

Chemoattraction of macrophages and T cells into the normal endometrium and inflammatory sites within endometriotic foci is mediated by chemokine gene expression. mRNA transcripts encoding regulated on activation, normal T-cell-expressed and -secreted (RANTES), a monocyte and T-cell chemokine, were demonstrated in the stroma of normal endometrium and endometriotic implants using in situ mRNA hybridization, Epithelial glands failed to express RANTES mRNA. In histological serial sections, we observed CD68-positive macrophages in the stroma of endometriotic implants adjacent to regions with prominent RANTES mRNA hybridization, In adjacent sections, monoclonal antibodies against tumor necrosis factor (TNF)-alpha showed this cytokine to be localized to stromal and epithelial compartments of the endometriotic implant with weak staining in unaffected ovarian tissue, Subconfluent monolayers of endometriotic stromal cells were tested for RANTES gene expression in situ, but we could only detect RANTES mRNA in isolated stromal cells after treatment with TNF-alpha. No RANTES mRNA was observed in unstimulated stromal cells or TNF-alpha stimulated or unstimulated epithelial cells, The data are consistent with a model in which proinflammatory cytokines (eg, TNF-alpha) induce RANTES gene expression limited to specific cells within endometrial and endometriotic stroma, Production of this chemokine, in turn, stimulates recruitment of CD68-positive macrophages into these tissues.

关键词：抗原 CD/分析抗原分化髓单核细胞/分析自身分泌性细胞间通讯细胞培养的趋化因子CCL5/生物合成趋化因子CCL5/遗传学趋化因子CCL5/免疫学子宫内膜异位症/代谢子宫内膜/代谢上皮细胞/药物作用上皮细胞/代谢免疫组织化学原位杂交巨噬细胞/化学旁分泌细胞细胞间通讯 RNA 信使/生物合成间质细胞/药物作用间质细胞/代谢肿瘤坏死因子α/药理学女(雌)性人类

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