The expression of cyclooxygenase-1 and -2 in proliferative endometrium and endometrial adenocarcinoma

BACKGROUND: The activity of cyclooxygenase-2 (COX-2) is increased in inflammation and in several cancer types. We investigated the expression of COX-2, cyclooxygenase-1 (COX-1), nitric oxide synthase-2 (NOS-2) and nitric oxide synthase-3 (NOS-3) in normal proliferative and secretory human endometrium, and in endometrial adenocarcinoma. METHODS: Human endometrium was collected at hysterectomy. Seven samples were in proliferative and 11 samples in secretory stage. Twelve specimens from endometrial carcinoma were collected, as well. Immunohistochemistry was used to investigate the expression of COX-1, COX-2, NOS-2 and NOS-3. RESULTS: COX-2 immunostaining was detected in most specimens of normal proliferative glandular epithelium (86%) and of endometrial carcinomas (92%). COX-2 staining was often detected in cancer cells on the border areas of the tumour and on the areas of invasive growth. Staining for COX-2 was seen in proliferative glands usually only in the basal layer of the endometrium. NOS-2 was usually absent or negligible in proliferative endometrial glands and also in the cancer cells of endometrial adenocarcinomas. No staining for either COX-2 or NOS-2 was seen in specimens of secretory glandular epithelium. The expression of the constitutive COX-1 and NOS-3 was negligible or weak in the glandular epithelium of proliferative and secretory endometrium and in endometrial cancer cells. CONCLUSIONS: The expression of the inducible COX-2 but not of COX-1 is stimulated in the glandular epithelium of proliferative endometrium and in the cancer cells of human endometrial adenocarcinoma, in particular in those in the borders of carcinoma and spreading into lymphatic vessels.     

Localization of the angiotensin II and its receptor subtype expression in human endometrium and identification of a novel high-affinity angiotensin II binding site.

Angiotensin (ANG) II is not only a potent vasoconstrictor but may also be involved in the regeneration of new blood vessels. In proliferative endometrium, ANG II-like immunoreactivity was detected in glandular epithelium and stroma with negligible staining around the vascular endothelium. In contrast, in secretory endometrium intense immunostaining was seen in the perivascular stromal cells around the endometrial spiral arterioles with negligible staining of the other cell types. Quantitative receptor autoradiography using the nonselective radioligand [125I]-ANG II and subtype selective competing compounds showed that endometrium contained predominantly AT2 receptors, with relatively low expression of AT1 receptors and a novel non-AT1/non-AT2 angiotensin II recognition site that was insensitive to AT1 or AT2 selective ligands. Levels of specific [125I]-ANG II receptor binding displayed cyclic changes during the menstrual cycle, reaching a maximum in early secretory endometrium and then decreasing in mid to late secretory endometrium to levels seen in early to mid proliferative endometrium. In situ hybridization showed AT1 receptor mRNA expression in the glands and in the endometrial blood vessels. The cyclic changes in ANG II-like immunoreactivity together with expression of both the known and the novel AT receptor subtypes imply that this octopeptide may play a dual role both in the control of the uterine vascular bed and also in the regeneration of the endometrium after endometrial shedding, acting as an angiogenic and mitogenic mediator.     

血管内皮生长因子及环氧合酶-2在子宫内膜腺癌中表达及意义

目的探讨血管内皮生长因子（vascular endothelial growth factor,VEGF）和环氧合酶-2（cyclooxygenase-2,COX-2）在子宫内膜腺癌中表达及临床意义。方法萎缩性正常子宫内膜患者12例（对照组）,子宫内膜增生症患者14例（子宫内膜增生组）,子宫内膜腺癌患者30例（子宫内膜腺癌组）,3组均手术切除子宫内膜组织标本,采用RT-PCR法检测子宫内膜组织COX-2及VEGF mRNA相对表达量。结果子宫内膜腺癌组、子宫内膜增生组、对照组中COX-2mRNA相对表达量分别为（133.58±56.96）%、（56.68±41.75）%、（8.33±5.21）%,VEGF mRNA相对表达量分别为（143.28±59.86）%、（57.33±46.78）%、（15.31±10.86）%,各组间COX-2mRNA及VEGF mRNA相对表达量比较差异有统计学意义（P〈0.01）;子宫内膜腺癌组COX-2表达与VEGF表达呈正相关（r=0.289,P=0.014）。结论COX-2及VEGF在子宫内膜腺癌中均呈高表达,其作为肿瘤恶化程度标记物可为临床诊疗工作提供参考依据。     

子宫内膜癌中Glut-1、C—erbB-2表达及其相互关系

目的探讨C—erbB-2和葡萄糖转运蛋白1（Glut-1）在不典型增生子宫内膜和子宫内膜腺癌中的表达特点及其相互关系。方法应用免疫组织化学方法检测Glut-1、C-erbB-2在30例正常增生期子宫内膜、32例不典型增生子宫内膜和35例子宫内膜腺癌中的表达,并结合临床资料进行分析。结果Glut-1、C—erbB-2在各组中的阳性表达率增生期子宫内膜均为0,不典型增生组分别为46．9％和59．4％,子宫内膜腺癌组82．9％,74．3％;二者在各试验组间均有显著差异（P〈0．005）。不典型增生组和子宫内膜腺癌组间差异有显著性（P〈0．005）。Glut-1与C-erbB-2在各组的表达呈显著正相关关系（P〈0．01）。结论Glut-1和C—erbB-2在子宫内膜腺癌的发生、发展中起着协同而独立的作用,对其进行联合检测有助于子宫内膜腺癌及癌前病变的早期诊断。     

子宫腺肌病中细胞凋亡和增殖状况及其相关基因表达研究

目的:探讨子宫腺肌病组织中细胞凋亡和增殖状况,及其相关调控基因survivin和PCNA在其中表达的意义。方法:应用免疫组化SP法,检测了30例子宫腺肌病患者的在位和异位子宫内膜及25例正常子宫内膜组织中survivin和PCNA的表达。结果:survivin蛋白在正常子宫内膜及腺肌病在位内膜分泌晚期的腺上皮细胞内有表达,而在增殖期无表达。在子宫腺肌病月经各期的异位内膜中,survivin蛋白持续表达,无周期性变化,其表达高于在位内膜及对照组内膜差异有显著性(P<0.05)。PCNA组织学评分在子宫腺肌病异位内膜,在位内膜以及对照组内膜,三组腺上皮细胞间有显著差异(P<0.05)。结论:腺肌病子宫内膜细胞凋亡减弱的同时增殖能力增强,可能是子宫腺肌病的发病机理之一。     

激活素A、抑制素A在正常子宫内膜的表达

目的：探讨激活素A（ACTA）、抑制素A（INHA）在正常子宫内膜的表达及其临床意义。方法30例患者的正常子宫内膜组织分为增生期16例（增生期组）、分泌期14例（分泌期组）。采用免疫组织化学链霉素抗生物素蛋白-过氧化物酶连结法检测两组子宫内膜ACTA、INHA的表达；电化学发光法检测血清性激素雌二醇（E2）、孕酮（P）水平；酶联免疫吸附测定法检测血清ACTA、INHA水平。结果 ACTA、INHA表达阳性细胞的着色部位主要是在子宫内膜腺体上皮细胞的细胞浆内，呈淡黄色、深棕色或褐色。分泌期组ACTA、INHA的表达均强于增生期，差异有统计学意义（t分别=-5.16、-4.19， P均＜0.05）。增生期组ACTA、INHA 表达水平与血清E2均呈正相关（r分别=0.53、0.61， P均＜0.05）；与血清P无相关性（r分别=-0.19、-0.30，P均＞0.05）；分泌期组ACTA 、INHA表达水平与血清E2、P均呈正相关（r分别=0.54、0.74；0.54、0.68，P均＜0.05）。增生期组和分泌期组ACTA、INHA 表达水平均与血清ACTA、INHA呈正相关（r分别=0.52、0.57；0.59、0.69，P均＜0.05）。结论 ACTA、INHA在正常子宫内膜增生期分泌期均有表达，但分泌期表达明显增强。 ACTA、INHA表达与血清E2、P密切相关，可能参与子宫内膜的成熟，在旁分泌/自分泌机制中起作用。     

PTEN 在子宫内膜增生过长和子宫内膜腺癌中的表达和临床意义

目的检测抑癌基因PTEN在子宫内膜腺癌中的表达,探讨其临床意义及在子宫内膜腺癌发生发展中的作用。方法应用免疫组化法检测10例增生期子宫内膜、10例单纯型增生过长、10例复杂型增生过长,10例非典型增生过长和46例子宫内膜腺癌组织中PTEN蛋白表达。结果PTEN在子宫内膜癌和不典型增生过长中的表达与单纯型 复杂型增生过长和增生期子宫内膜比较差异有显著性。46例子宫内膜样腺癌中PTEN的阴性率是52·2%,PTEN阴性表达与子宫内膜腺癌的细胞分化程度、肌层浸润有关,有统计学意义(P<0·05)。结论PTEN表达缺失在子宫内膜癌发生、发展中起一定的作用,和子宫内膜腺癌的生物学行为有关,测定PTEN的表达对评估子宫内膜癌预后有重要意义,可作为子宫内膜腺癌预后的生物学检测指标。     

子宫内膜异位症细胞增殖与凋亡探讨

目的 探讨子宫内膜异位症的细胞增殖和细胞凋亡。方法 应用免疫组化法对61例正常月经周期子宫内膜及卵巢子宫内膜异位症患者进行PCNA检测。同时用TUNEL法对其中34例标本进行凋亡检测。结果 正常位置的子宫内膜分泌期及月经期凋亡细胞较多，增生期内膜及异位内膜无凋亡。PCNA蛋白在增生期含量高，分泌期低落，月经期消失，异位内膜中高于增生期（P〈0．05）。结论 凋亡与子宫内膜脱落有关；异位子宫内膜细胞通过过度增殖且减少凋亡，延长生命周期。     

环氧合酶-2在卵巢癌组织中的表达

目的:探讨环氧合酶-2(cyclooxygenase-2,COX-2)在卵巢癌组织中的表达及意义。方法:应用免疫组化和RT-PCR技术检测正常卵巢和卵巢癌组织中COX-2的表达情况。结果:检测的38例卵巢癌组织COX-2蛋白表达阳性23例,阳性表达率61%,33例表达COX-2mRNA,阳性表达率为87%;20例正常卵巢组织COX-2蛋白与COX-2mRNA均呈阴性表达。结论:COX-2在卵巢癌组织高表达,并在其发生发展中起重要作用。     

环氧化酶-1在乳腺癌中的表达及其意义

目的:探讨环氧化酶-1(cyclooxygenase-1,COX-1)在乳腺癌中的表达与临床病理指标之间的相关性,及其对肿瘤血管生成的影响。方法:应用免疫组织化学方法检测61例乳腺癌组织标本的COX-1和血管内皮生长因子(VEGF)的表达。结果:61例乳腺癌组织标本中COX-1阳性率为60.7%,COX-1和VEGF表达明显相关(P=0.026)。在肿块<2cm组以及临床Ⅰ期组中COX-1的表达明显增强(P<0.05)。结论:乳腺癌组织中存在COX-1的高表达,COX-1在乳腺癌的早期阶段可能起着重要作用,VEGF可能是COX-1介导肿瘤新生血管形成的重要因子之一,为非甾体类抗炎药物应用于乳腺癌的预防提供了依据。     

Quantitative evaluation of the activity of nuclear Ca2+/Mg2+-dependent endonucleases in hyperplasia and cancer of the endometrium

Endometrial carcinoma is the most incident cancer of human reproductive system. There are unequivocal evidences of relationship between complex and atypical hyperplasia and development of cancer. Apoptosis plays a significant role in the maintenance of equilibrium between cell death and proliferation and contributes to prevention of tumorigenesis. Internucleosomal DNA fragmentation known as one of the most important criteria of apoptosis cannot be used for evaluating the risk of cancer development because it reflects the current level of apoptosis but is useless for evaluating the real limits of apoptosis intensity in certain types of tissue. For estimating the possibility of apoptosis development in endometrial tissues, a new method of quantitation of nuclear Ca(2+)/Mg(2+)-dependent endonuclease (NCME) activity has been developed. Fifteen samples of normal endometrial tissues at middle proliferative, secretory, premenstrual, and menstrual phases, 43 samples of hyperplastic endometrial tissues, 13 samples of endometrial polyps, and 17 samples of endometrial adenocarcinoma were collected by diagnostic curettage of the uterine cavity and by hysterectomy (carcinomas). The material was examined by 1) TUNEL method and 2) agarose gel electrophoresis of DNA cleaved by nuclear CME in isolated cell nuclei in the presence of Ca(2+) and Mg(2+) ions, followed by quantitation of CME activity. The activity of NCME was found to decrease from normal endometrium (1.1 +/- 0.12 U, without significant changes throughout the menstrual cycle) through polyps (0.9 +/- 0.15 U), cystic hyperplasia (0.45 +/- 0.06 U, p < 0.01), and adenomatous hyperplasia (0.32 +/- 0.08 U, p < 0.01) to adenocarcinoma (0.37 +/- 0.11 U, p < 0.01 for well differentiated, 0.16 +/- 0.08 U, p < 0.01 for moderately differentiated, and 0.03 +/- 0.02 U, p < 0.01 for poorly differentiated samples). The TUNEL-specific staining pattern in normal endometrium varied in a wide range during the menstrual cycle (from poorly stained individual cells in the proliferative phase to intensely stained cell clusters in the premenstrual phase). At the same time the difference between the normal endometrium in the proliferative phase and pathologically changed endometrium (hyperplasia or cancer) could not be detected by the TUNEL technique. Hence, TUNEL is useless for predicting cancer development in hyperplasia and precancer. By contrast, evaluation of NCME activity helps detect the early disorders in the proliferative processes coursing in endometrial tissues and thus prevent tumor development.     

DKK1在雌孕激素受体阴性子宫内膜癌中高表达及意义

目的 探讨DKK1在不同雌孕激素受体状态的子宫内膜癌组织中的表达及意义.方法 采用免疫组化检测DKK1在子宫内膜癌及正常子宫内膜组织中的表达及定位;比较DKK1表达水平在不同雌孕激素受体状态子宫内膜癌组织及正常子宫内膜组织的差异,并探讨其与子宫内膜癌临床病理特征之间的关系.结果 免疫组化结果显示DKK1蛋白主要表达于子宫内膜癌腺上皮上,基质中少量表达.DKK1在雌孕激素受体阴性子宫内膜癌组织中的表达水平高于雌或孕激素受体阳性组和正常子宫内膜组,差异具有统计学意义（P＜0.05）.DKK1蛋白表达水平与雌孕激素受体状态、有无淋巴结转移密切相关（P＜0.05）,而与患者年龄、病理分级、分期无关（P＞0.05）.结论 DKK1在雌孕激素受体阴性子宫内膜癌组织中高表达.     

三叶因子2和环氧合酶-2在胃癌组织中的表达

目的:研究三叶因子2(TFF2)和环氧合酶-2(COX-2)在正常胃黏膜组织、癌旁组织和胃癌组织中的表达情况,探讨TFF2表达与胃癌发生的关系。方法:采用SP免疫组化方法检测30例正常胃黏膜组织、50例癌旁组织和50例胃癌组织中TFF2和COX-2的表达。结果:正常胃黏膜组织→癌旁组织→胃癌组织中,TFF2表达呈逐渐减弱趋势(165.80±16.42,184.44±19.02和206.79±17.62,P<0.01),而COX-2的表达呈逐渐上升趋势(217.50±12.60,203.08±17.68和193.50±16.160,P<0.01),TFF2的表达强度与COX-2呈负相关(r=-0.761,P<0.01)。结论:COX-2的表达增加抑制TFF2的表达,细胞增殖与凋亡失衡,最终导致胃癌的发生。     

17 beta-Hydroxysteroid dehydrogenase type 2: chromosomal assignment and progestin regulation of gene expression in human endometrium.

The cDNAs for two separate human 17 beta-hydroxysteroid dehydrogenases (17 beta-HSD) have been isolated and sequenced. The well-studied human placental cytosolic 17 beta-HSD (also referred to as estradiol dehydrogenase) preferentially catalyzes the reduction of estrone to estradiol-17 beta and the reduction of the C-20-ketone of progesterone to 20 alpha-dihydroprogesterone. This isoform of the enzyme has been referred to as 17 beta-HSD type 1 and localized to chromosome 17. A second 17 beta-HSD isoform (referred to as type 2) is localized in the endoplasmic reticulum of human trophoblast and is characterized by the preferential oxidation of the C-17 beta-hydroxyl group of C18- and C19-steroids and the C-20 alpha-hydroxyl group of 20 alpha-dihydroprogesterone. In this study, we determined the chromosomal localization of human 17 beta-HSD type 2, the expression of this gene in human endometrium, and the tissue distribution of the mRNA. We found that the human 17 beta-HSD type 2 gene is localized on chromosome 16, 16q24. 17 beta-HSD type 2 mRNA (approximately 1.5 kb) was identified in human endometrial tissues by Northern analysis of total RNA (10 micrograms). The highest levels of 17 beta-HSD type 2 mRNA were found in endometrial tissues obtained during the mid- to late secretory phase of the ovarian cycle (i.e., during the time of high plasma levels of progesterone). 17 beta-HSD type 2 mRNA levels were much greater in glandular epithelium than in the stromal cells isolated from secretory phase endometrium. The levels of 17 beta-HSD type 2 mRNA in secretory phase endometrium were approximately one-tenth that in villous trophoblast tissue from human placenta. We did not detect 17 beta-HSD type 1 mRNA in endometrial tissue by Northern analysis of total (10 micrograms) RNA. These findings are consistent with the view that the progestin-regulated 17 beta-HSD of the glandular epithelium of the human endometrium is primarily, if not exclusively, the product of the 17 beta-HSD type 2 gene. 17 beta-HSD type 2 mRNA was present in human placenta, liver, and small intestine; much smaller amounts, barely detectable by Northern analysis of poly(A)+ RNA, were present in prostate, kidney, pancreas, and colon, but not in heart, brain, skeletal muscle, spleen, thymus, ovary, or testis.     

胃泌素、环氧合酶2在不同胃黏膜病变中的表达及意义

目的 研究不同胃黏膜病变组织中胃泌素、环氧合酶2(COX-2)和Ki-67的表达,探讨胃泌素和COX-2在胃黏膜癌变过程中的作用及其相互关系.方法 96例不同胃黏膜病变组织来自胃镜活检标本.采用免疫组化染色EnVision法检测胃泌素、COX-2和Ki-67表达.结果 胃泌素在慢性浅表性胃炎(CSG)、慢性萎缩性胃炎(CAG)、肠上皮化生(IM)、异型增生(Dy)和胃癌(GC)中表达的阳性率分别为58.8%、30.0%、57.9%、68.2%和77.8%,CAG组与GC组比较,差异有统计学意义(P<0.05).COX-2在CSG、CAG、IM、Dy和GC中表达的阳性率分别为41.2%、45.0%、68.4%、72.7%和88.9%;在GC中的表达显著高于CSG(P<0.05).从CSG→GC,Ki-67增殖指数(PI)呈逐渐递增趋势;从CAD→GC,胃泌素和COX-2的表达与PI呈正相关(P<0.05).在胃癌前期病变中,胃泌素与COX-2表达有显著相关性(γ=0.4108,P<0.01),二者共同表达时,PI显著升高.结论 CAG中胃泌素王低表达,从CAG→GC,胃泌素表达逐渐升高.在胃黏膜癌变过程中,COX-2表达呈逐渐递增趋势.在胃癌前病变阶段,胃泌素和COX-2协同促进细胞增殖,胃泌素可能参与上调COX-2的表达.     

生存素基因在子宫内膜异位症中的表达及促性腺激素释放激素激动剂、环氧合酶-2抑制剂对其表达的影响

目的 探讨生存素(survivin)基因在子宫内膜异位症(FMs)发病机制中的作用.方法 检测生存素在正常与异位子宫内膜组织中的表达;观察促性腺激素释放激素激动剂(GnRHa)、环氧合酶-2(COX-2)抑制剂对体外培养的EMs异位内膜细胞及正常子宫内膜细胞中survivin基因mRNA表达的调节作用及对体外培养的异位内膜凋亡率的影响.结果 ①EMs异位组、在位组survivin mRNA的表达明显强于对照组(均P<0.05),且无周期性变化.②GnRHa可呈浓度依赖性下调体外培养的异位内膜细胞及正常子宫内膜细胞中survivin mRNA的表达,COX-2抑制剂亦呈浓度依赖性下调体外培养的异位内膜细胞及正常子宫内膜细胞中survivln mRNA的表达,GnRHa100μL加COX-2抑制荆40 μmol/L可以促进体外培养的异位内膜细胞凋亡(P<0.05),二者无明显协同作用(P>0.05)结论 ①异位内膜细胞高表达survivin,对凋亡的敏感性低,使异位灶存活并发展;②GnRHa、COX-2抑制剂可通过抑制survivin的表达来促进体外培养的EMs异位内膜细胞的凋亡.     

Involvement of granule-mediated apoptosis in the cyclic changes of the normal human endometrium

Our objective is to investigate the involvement of granule-mediated apoptosis in the cyclic changes of the endometrium. We demonstrated the localization of CD56, perforin, granzyme B and caspase-3 in the endometrium by immunohistochemistry. We also confirmed the localization of perforin by immuno-electron microscopy, and demonstrated apoptosis in endometrial glandular cells by TdT-mediated dUTP-biotin nick end labeling (TUNEL) and electron microscopy. Uterine CD56-positive natural killer (NK) cells expressed perforin and granzyme B in its cytoplasm. Uterine NK cells increased significantly in the endometrial stroma during the secretory phase, and peaked during the late secretory phase. These cells started decreasing in number during the menstrual period. In endometrial glandular cells, caspase-3 and TUNEL-positive cells increased significantly from the late secretory phase, with apoptosis reaching a peak during the menstrual period. Using electron microscopy, we observed uterine NK cells with chromatin rich, segmented nuclei and intracytoplasmic granules in the stroma obtained from late secretory phase endometria. These cells extended projections to the lining of endometrial glandular cells and attached to form a cell-to-cell contact. In addition, nuclear chromatin was observed to have already cohered and small cytoplasmic organelles were beginning to disappear, suggesting that these endometrial glandular cells were undergoing apoptosis. Utilizing immuno-electron microscopy, intracytoplasmic granules in uterine NK cells were stained with anti-perforin antibody. The findings of this study suggest that granule-mediated apoptosis in endometrial glandular cells induced by NK cells expressing perforin and granzyme B may be associated with the onset of menstruation.     

塞来昔布对肺癌细胞的增殖抑制作用研究

目的研究非甾体类抗炎药塞来昔布对肺癌细胞的体外抑制作用。方法选用肺癌A549细胞系体外培养,在不同塞来昔布浓度作用下,MTT法检测肺癌细胞的增殖抑制,流式细胞仪测定肺癌细胞周期分布以及RTPCR法观察肺癌细胞血管内皮生长因子mRNA(VEGFmRNA)的表达水平。结果塞来昔布对肺癌细胞增殖有抑制作用,且呈剂量依赖关系,IC50为50μmol/L;塞来昔布可将肺癌细胞阻滞在G0/G1期,有剂量依赖性;RTPCR法中,对照组肺癌细胞VEGFmRNA的表达水平高于药物组(P<0.05),药物组中肺癌细胞VEGFmRNA表达水平与塞来昔布浓度相关。结论塞来昔布对肺癌细胞有明显的体外增殖抑制作用,与VEGFmRNA的表达下调和阻滞细胞周期在G/G期有关。