神经激肽B受体在小鼠消化道内的定位

采用免疫组织化学ABC染色方法研究了神经激肽B受体（NK3r)在小鼠消化道的分布。MK3r样阳性的神经无胞体及神经纤维主要分布在十二指肠,空肠,回肠及结肠的粘膜下层神经丛和肌间神经丛,NK3r样阳性产物在食管,胃和直肠的神经丛中未见分布;NK3r样阳性产物大部分避限于神经细胞表面,也存在于胞和一些轴突内部,并在胞质中较细胞表面染色浅。。统计结果表明NK3r样免疫阳性神经元占肠神经系统神经元总数的0．5－1％,提示小鼠消化道内NK3r样阳性神经元可能参与消化功能的调节。     

UHRF1在结肠癌中的表达及其与细胞增殖的关系

目的:探讨UHRF1(Ubiquitin-like with plant homeodomain and ring覱nger domains 1)在结肠癌组织中的表达,及其与结肠癌细胞增殖的关系。方法:采用免疫组织化学方法检测62例结肠癌组织和癌旁正常对照组织的UHRF1表达,统计分析其表达和临床病理资料和预后关系。采用si RNA抑制UHRF1表达,分析其与细胞增殖的关系。结果:结肠癌组织中UHRF1高表达,癌旁正常组织低表达。62例结肠癌组织中33例UHRF1阳性,阳性率为53.2%。UHRF1表达与淋巴结转移、远处器官转移和Dukes'分期相关(P0.05)。高表达UHRF1结肠患者的生存时间明显较低表达者短(P0.05)。单因素分析显示淋巴结转移、远处器官转移、UHRF1和Dukes'分期与患者累积生存率相关(P0.05)。多因素分析显示UHRF1可作为结肠癌患者预后的独立危险因子(P0.05)。采用si RNA转染HCT116后,UHRF1表达显著降低,细胞增殖被抑制。结论:UHRF1参与结肠癌细胞增殖,与结肠癌患者预后密切相关。     

交感神经节细胞内P物质受体与5—羟色胺受体的共存及其联系

应用离体神经节细胞内记录,观察到在１３３个鼠腹腔神经节细胞及１２９个肠系膜下社会节细胞中,分别有６６个细胞及４７个细胞对Ｐ物质及５－羟色胺同时敏感,分别有４０个细胞及６５个细胞仅对其中一种递质敏感,另有２７个细胞及１７个细胞对其中一种递质敏感。     

P物质受体在大鼠纹状体边缘区内的表达

我们以前的工作观察到纹状体边缘区内有密集的P物质纤维及终末分布,本用原位杂交和免疫组织化学方法研究了大鼠纹状体边缘区内P物质受体（SPR）的表达及分布,原位杂交结果发现P物质mRNA阳性杂交信号在纹状体内的分布不均匀,尾壳核内只有少量中等大小的阳性胞体,苍白球内只有少量较大的阳性胸体,而在尾壳核和苍白球之间的边缘区部位则可见许多中等大小的梭形阳性神经元胞体,并呈现密集的带关分布。免疫组织化学结果观察到P物质阳性神经元胞体在纹状体内的分布与原位杂交结果一致。推测大鼠纹体边缘区内可以合成P物质受体,具有接受和整合P物质神经递质的功能,推测边缘区内SPR神经元可能对SP递质的接受、调节有重要作用。     

神经激肽-1(NK-1)受体介导福尔马林诱发的大鼠脊髓c-fos表达

本文用Ｆｏｓ作为背角伤害性反应神经元活动的一个标志物,结合免疫细胞化学和神经药理学方法,观察了速激肽受体拮抗剂对福尔马林诱发的脊髓ｃ－ｆｏｓ原癌基因表达的影响。一侧大鼠后肢跖部皮下注射福尔马林,仅在同侧脊髓背角有ｃ－ｆｏｓ表达。Ｆｏｓ阳性神经元最密集分布于Ｉ层和Ⅱ层背侧的内侧部,中等量分布于Ⅳ层和Ｖ型,少量定位于Ⅱ层腹侧部、Ⅲ、Ⅵ和Ⅹ层。若预先在一侧大鼠后肢跖部皮下注射福尔马林前,鞘内给予神经激肽     

P63和TTF-1在肺癌组织中的表达及意义

目的研究P63和TTF-1在肺癌各种类型组织中的表达及其意义。方法随机收集原发性肺癌组织标本53例（其中鳞癌16例,腺癌16例,小细胞癌14例,大细胞癌7例,均为中低分化程度癌组织）,采用免疫组织化学S-P法分别检测P63蛋白和TTF-1蛋白在各种类型肺癌组织中的表达并结合二者表达的结果进行分析。结果二者的表达在肺癌细胞中定位于细胞核,呈棕黄色。在肺鳞癌、肺腺癌、肺小细胞癌和肺大细胞癌中,P63的阳性表达率分别为93.8%（15/16）、37.5%（6/16）、21.4%（3/14）、28.6%（2/7）;TTF-1的阳性表达率分别为18.8%（3/16）、75%（12/16）、78.6%（11/14）、0%（0/7）。结论①P63在肺鳞癌中的表达水平较高,可以作为鉴别低分化鳞癌与低分化腺癌,小细胞癌的指标;②TTF-1在低分化腺癌和小细胞癌中的表达水平较高,对于肺癌组织类型和非癌组织的鉴别具有一定意义;③根据P63和TTF-1在肺癌组织的特异性表达,将二者联合起来有助于对低分化鳞癌和低分化腺癌以及低分化鳞癌和小细胞癌的鉴别诊断。     

PBRM1 和P21 蛋白在子宫内膜癌的表达及意义

目的：探讨PBRM1 和P21 蛋白在子宫内膜癌中的表达和临床意义。方法：应用免疫组化方法对105 例子宫内膜癌组织进 行PBRM1和P21 蛋白检测。结果：子宫内膜癌中PBRM1 和P21 的阳性表达率分别为62.86%和80%;癌旁组织PBRM1 和P21 阳性表达率分别为45.71%和34.2%,子宫内膜癌组织表达高于癌旁组织（P＜0.05）;PBRM1 和P53 阳性表达与组织学类型、浸润 深度、淋巴结转移、组织学分级、临床TNM分期相关（P＜0.05）,与患者年龄无关（P＞0.05）。PBRM1 和P21 的表达呈正相关（P＜ 0.05）。结论：PBRM1 和P21 表达水平与子宫内膜癌组织的病理分级和临床分期的增高而增高,对其进行监测对子宫内膜癌的诊 断、治疗及预后有指导意义。     

Involvement of substance P and the NK-1 receptor in cancer progression

Many data suggest the deep involvement of the substance P (SP)/neurokinin (NK)-1 receptor system in cancer: (1) Tumor cells express SP, NK-1 receptors and mRNA for the tachykinin NK-1 receptor; (2) Several isoforms of the NK-1 receptor are expressed in tumor cells; (3) the NK-1 receptor is involved in the viability of tumor cells; (4) NK-1 receptors are overexpressed in tumor cells in comparison with normal ones and malignant tissues express more NK-1 receptors than benign tissues; (5) Tumor cells expressing the most malignant phenotypes show an increased percentage of NK-1 receptor expression; (6) The expression of preprotachykinin A is increased in tumor cells in comparison with the levels found in normal cells; (7) SP induces the proliferation and migration of tumor cells and stimulates angiogenesis by increasing the proliferation of endothelial cells; (8) NK-1 receptor antagonists elicit the inhibition of tumor cell growth; (9) The specific antitumor action of NK-1 receptor antagonists on tumor cells occurs through the NK-1 receptor; (10) Tumor cell death is due to apoptosis; (11) NK-1 receptor antagonists inhibit the migration of tumor cells and neoangiogenesis. The NK-1 receptor is a therapeutic target in cancer and NK-1 receptor antagonists could be considered as broad-spectrum antitumor drugs for the treatment of cancer. It seems that a common mechanism for cancer cell proliferation mediated by SP and the NK-1 receptor is triggered, as well as a common mechanism exerted by NK-1 receptor antagonists on tumor cells, i.e. apoptosis.     

Presence of substance P and the neurokinin-1 receptor in tenocytes of the human Achilles tendon

Nerve signal substances, such as the tachykinin substance P (SP), may be involved in the changes that occur in response to tendinopathy (tendinosis). It is previously known that the level of SP innervation within tendon tissue is limited, but results of experimental studies have suggested that SP may have stimulatory, angiogenetic and healing effects in injured tendons. Therefore, it would be of interest to know if there is a local SP-supply in tendon tissue. In the present study, the patterns of expression of SP and its preferred receptor, the neurokinin-1 receptor (NK-1 R), in normal and tendinosis human Achilles tendons were analyzed by use of both immunohistochemistry and in situ hybridization. We found that there was expression of SP mRNA in tenocytes, and that tenocytes showed expression of NK-1 R at protein as well as mRNA levels. The observations concerning both SP and NK-1 R were most evident for tenocytes in tendinosis tendons. Our findings suggest that SP is produced in tendinosis tendons, and furthermore that SP has marked effects on the tenocytes via the NK-1 R. It cannot be excluded that the SP effects are of importance concerning the processes of reorganization and healing that occur for tendon tissue in tendinosis. In conclusion, it appears as if SPergic autocrine/paracrine effects occur in tendon tissue during the processes of tendinosis, hitherto unknown effects for human tendons.     

A fragment of substance P with specific central activity: SP(1-7)

Amino-terminal fragments of substance P (SP), SP(1-7) and SP(1-8), were found to produce naloxone-reversible antinociception in the mouse similar to that produced by SP. Similar to SP, these peptides produce antinociception only within a narrow dose range. They have no activity on smooth muscle or blood pressure. These results suggest that contrary to peripheral effects of SP, which are mediated by receptors which recognize the carboxy-terminal part of the SP molecule, certain central actions of SP are mediated by receptors which recognize the amino-terminal part of the SP molecule. SP may be metabolized to this active fragment prior to its action at these receptors.     

The substance P/neurokinin-1 receptor system in lung cancer: Focus on the antitumor action of neurokinin-1 receptor antagonists

The last decades have seen no significant progress in extending the survival of lung cancer patients and there is an urgent need to improve current therapies. The substance P (SP)/neurokinin-1 receptor (NK-1R) system plays an important role in the development of cancer: SP and NK-1R antagonists respectively induce cell proliferation and inhibition in human cancer cell lines. No study of the involvement of this system in non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) cells has been carried out in depth. Here, we demonstrate the involvement of the SP/NK-1R system in human H-69 (SCLC) and COR-L23 (NSCLC) cell lines: (1) they express isoforms of the NK-1R and mRNA for the NK-1R; (2) they overexpress the tachykinin 1 gene; (3) the NK-1R is involved in their viability; (4) SP induces their proliferation; (5) NK-1R antagonists (Aprepitant (Emend), L-733,060, L-732,138) inhibit the growth of both cell lines in a concentration-dependent manner; (6) the specific antitumor action of these antagonists against such cells occurs through the NK-1R; and (7) lung cancer cell death is due to apoptosis. We also demonstrate the presence of NK-1Rs and SP in all the human SCLC and NSCLC samples studied. Our findings indicate that the NK-1R may be a promising new target in the treatment of lung cancer and that NK-1R antagonists could be new candidate antitumor drugs in the treatment of SCLC and NSCLC.     

Substance P and substance K receptor binding sites in the human gastrointestinal tract: Localization by autoradiography

Quantitative receptor autoradiography was used to localize and quantify the distribution of binding sites for ¹²⁵I-radiolabeled substance P (SP), substance K (SK) and neuromedin K (NK) in the human GI tract using histologically normal tissue obtained from uninvolved margins of resections for carcinoma. The distribution of SP and SK binding sites is different for each gastrointestinal (GI) segment examined. Specific SP binding sites are expressed by arterioles and venules, myenteric plexus, external circular muscle, external longitudinal muscle, muscularis mucosa, epithelial cells of the mucosa, and the germinal centers of lymph nodules. SK binding sites are distributed in a pattern distinct from SP binding sites and are localized to the external circular muscle, external longitudinal muscle, and the muscularis mucosa. Binding sites for NK were not detected in any part of the human GI tract. These results demonstrate that: 1) surgical specimens from the human GI tract can be effectively processed for quantitative receptor autoradiography; 2) of the three mammalian tachykinins tested, SP and SK, but not NK binding sites are expressed in detectable levels in the human GI tract; 3) whereas SK receptor binding sites are expressed almost exclusively by smooth muscle, SP binding sites are expressed by smooth muscle cells, arterioles, venules, epithelial cells of the mucosa and cells associated with lymph nodules; and 4) both SP and SK binding sites expressed by smooth muscle are more stable than SP binding sites expressed by blood vessels, lymph nodules, and mucosal cells.     

O-GlcNAcylation is a novel regulator of lung and colon cancer malignancy

O-GlcNAc is a monosaccharide attached to serine or threonine hydroxyl moieties on numerous nuclear and cytoplasmic proteins; O-GlcNAcylation is dynamically regulated by O-GlcNAc transferase (OGT) and O-GlcNAcase (OGA). Although recent studies have shown that O-GlcNAcylation plays essential roles in breast cancer progression, it is also necessary to know whether O-GlcNAcylation is involved in other types of human cancer. In this study, O-GlcNAcylation levels and the expressions of OGT and OGA in human lung and colon cancer tissues were examined by immunohistochemistry analysis. We found that O-GlcNAcylation as well as OGT expression was significantly elevated in the cancer tissues compared with that in the corresponding adjacent tissues. Additionally, the roles of O-GlcNAcylation in the malignancy of lung and colon cancer were investigated in vitro. The results showed that O-GlcNAcylation markedly enhanced the anchorage-independent growth of lung and colon cancer cells; O-GlcNAcylation could also enhance lung and colon cancer invasion in a context-dependent manner. All together, this study suggests that O-GlcNAcylation might play important roles in lung and colon cancer formation and progression, and may be a valuable target for diagnosis and therapy of cancer.     

长爪沙鼠肠道生长抑素和P物质细胞密度的年龄变化

胃肠激素对调节小哺乳动物的消化功能具有重要作用。本文应用卵白素- 生物素- 过氧化物酶复合物(Avidin-biotin-peroxidase complex,ABC)免疫组织化学法,对冬季幼年、成年和老年长爪沙鼠肠道生长抑素(Somtostatin,SS)和P 物质(Substance P,SP)细胞进行了定位和比较。结果显示:不同年龄组两种内分泌细胞的形态学特征相似,呈圆形、椭圆形、梭形、锥形或不规则形。SS 细胞随年龄增加而分布范围缩小,主要分布于小肠和十二指肠,老年鼠的结肠和直肠无分布;盲肠段老年鼠高于幼年鼠和成年鼠,其余各段均无年龄差异。幼年鼠、成年鼠和老年鼠SP 细胞分别以结肠、盲肠和直肠密度为最高,成年鼠结肠和直肠无分布;肠道各段的密度均有年龄差异。这些结果表明,长爪沙鼠肠道SS 和SP 细胞的分布模式和发育特点有年龄差异,这可能与其生存环境的食物质量和两种内分泌细胞相互拮抗的消化生理功能有关。     

The NK-1 Receptor and a Calcium-Phospholipid Pathway: Inositol Trisphosphate Production and Calcium Movements Induced by Selective Agonists of Neurokinin Receptors in Rat Parotid Glands

In the rat parotid gland, substance P has been shown to induce a phosphatidylinositol bisphosphate breakdown resulting in an inositol trisphosphate production. These data suggested that substance P activated a phospholipase C and thus mediated its effects through the calcium-phospholipid pathway. To determine which neurokinin (NK) receptor was involved in the substance P response, we have used selective agonists of the different NK receptors and examined their effects on both inositol trisphosphate production and calcium movements. A selective NK-1 receptor agonist, [Sar9Met(O2)11]-substance P, evoked an [3H]inositol trisphosphate production and a rapid and transient 45Ca2+ efflux. On the other hand, selective NK-2 and NK-3 receptor agonists, [beta-Ala8]-NKA(4-10) and [MePhe7]-NKB, respectively, were without effect. We conclude that, in the rat parotid glands, only the NK-1 receptors are coupled to the calcium-phospholipid pathway. The C-terminal part of substance P appeared to be sufficient to stimulate this route because the C-terminal octapeptide, substance P(4-11), mimicked substance P effects on both inositol trisphosphate production and calcium movements. The NK-2 and NK-3 receptors, if present in the rat parotid glands, are not associated with the calcium-phospholipid pathway.