Autocrine stimulation by insulin-like growth factor I is involved in the growth, tumorigenicity and chemoresistance of human esophageal carcinoma cells

Insulin-like growth factor I (IGF-I) receptor (IGF-IR)-mediated signals are known to be involved in cell growth and transformation and prevention of apoptosis. In this study, we demonstrated the coexpression of IGF-I and IGF-IR in human esophageal carcinoma tissues. We also demonstrated the IGF-I autocrine system in esophageal carcinoma cell lines. Both the CE48T/VGH and CE81T/VGH cell lines showed proliferative responses to IGF-I stimulation. Autokinase activity of IGF-IR in these cells can be triggered by the exogenous addition of IGF-I. In addition, an IGF-I peptide antagonist, JB1, specifically inhibited ligand-induced receptor autophosphorylation in a dose-dependent manner. Under serum-free conditions, JB1 also reduced the degree of IGF-IR phosphorylation and cell numbers. Furthermore, the addition of JB1 decreased the number of CE81T/VGH colonies formed in methyl cellulose agar and the size and the incidence of tumors which grew in mice with severe combined immunodeficiency. These results imply that an IGF-I autocrine system in human esophageal carcinoma cells could stimulate tumor growth. Finally, we found that IGF-I prevented the apoptosis of CE81T/VGH cells induced by chemotherapeutic drugs, such as cisplatin, 5-fluorouracil and camptothecin. Thus, interruption of IGF-IR function may provide a way to retard tumor growth and increase the sensitivity of esophageal carcinoma to chemotherapy.     

An Immunofluorescent Method for Characterization of Barrett’s Esophagus Cells

Esophageal adenocarcinoma (EAC) has an overall survival rate of less than 17% and incidence of EAC has risen dramatically over the past two decades. One of the primary risk factors of EAC is Barrett’s esophagus (BE), a metaplastic change of the normal squamous esophagus in response to chronic heartburn. Despite the well-established connection between EAC and BE, interrogation of the molecular events, particularly altered signaling pathways involving progression of BE to EAC, are poorly understood. Much of this is due to the lack of suitable in vitro models available to study these diseases. Recently, immortalized BE cell lines have become commercially available allowing for in vitro studies of BE. Here, we present a method for immunofluorescent staining of immortalized BE cell lines, allowing in vitro characterization of cell signaling and structure after exposure to therapeutic compounds. Application of these techniques will help develop insight into the mechanisms involved in BE to EAC progression and provide potential avenues for treatment and prevention of EAC.     

益生菌联合肠内营养对 食管癌患者术后胃肠功能的影响

目的探讨益生菌联合肠内营养对食管癌患者术后胃肠功能的影响。方法 50例食管癌患者按随机数字表法被分为对照组(24例)和试验组(26例)。对照组患者采用常规治疗方案,并同时实施肠内营养治疗。试验组患者在对照组治疗的基础上给予鼻饲益生菌。观察并比较两组患者腹胀、腹泻、便秘、胃内容物反流、胃潴留等不良反应的发生情况;留置胃管、留置鼻肠营养管及达到肠内营养目标喂养量的时间;患者入院时和出院时体重的变化以及两组患者住院时间及住院费用。结果试验组患者腹胀、便秘、胃潴留的发生率均明显低于对照组,差异有统计学意义(P0.05)。两组患者腹泻、胃内容物反流的发生率差异无统计学意义(P0.05)。试验组患者达到肠内营养目标喂养量的时间显著短于对照组(P0.05)。试验组患者留置胃管时间和留置鼻肠营养管的时间明显短于对照组(P0.05)。对照组患者出院时体重较入院时明显降低,与试验组相比差异有统计学意义(P0.05)。试验组患者的住院时间和住院费用均低于对照组,但差异无统计学意义(P0.05)。结论肠内营养联合益生菌具有协同治疗作用,可有效降低食管癌患者术后胃肠道不良反应的发生率,促进患者肠内营养的实施,保证患者机体的营养需要。     

Morphology and immunocytochemistry of two endocrine cell types in the guinea-pig esophageal epithelium

Summary Two types of endocrine cells in the basal layer of the keratinized stratified squamous epithelium in the guinea-pig esophagus were studied with immunohistochemistry by means of a streptavidin-biotin-bridge technique and by the immunofluorescence double-labelling technique. Cell-type I exhibited immunoreactivity to chromogranin A and to nucleosidetriphosphate-adenosinediphosphate(ADP)-phosphotransferase. Ultrastructurally, this cell type was characterized by small cytoplasmic dense-core vesicles in which immunoreactive product was localized. Cell-type II contained large membrane-limited granules, which were moderately electron dense. These granules displayed somatostatin immunoreactivity. Both cell types were located in close vicinity to non-myelinated nerve fibers and small blood vessels. The results provide evidence that, independent from the type of lining epithelium, the gastro-enteropancreatic system in guinea-pig extends to the lower portion of the esophagus.Supported by the German Research Foundation, grant He 919/6-2     

三种术式重建食管治疗食管癌和贲门癌的临床研究

目的:提高食管癌和贲门癌的根治切除率和临床治愈率,预防吻合口瘘。方法:设计了Ⅰ、Ⅱ、Ⅲ三种术式:上、中段食管癌采用右胸前外侧切口,经第3或4肋间开胸,左腹直肌旁或左肋弓下斜切口开腹和右颈部切口,保留2～3cm颈段食管,食管次全切除,贲门部或部分胃切除,在颈部食管胃端侧分层吻合(Ⅰ式)。中段食管癌采用左胸前外侧切口,经第4或第5肋间开胸,腹部切口同前,左颈部切口,保留3～4cm颈段食管,食管次全切除,部分胃切除,在左颈部食管胃端侧分层吻合(Ⅱ式)。贲门癌采用左胸前外侧切口,经第5或4肋间开胸,腹部切口同前,中段食管和近半胃或纵半胃切除,在主动脉弓下食管胃端侧分层吻合(Ⅲ式)。尽可能清除区域淋巴结,吻合口均用大网膜包盖加固。结果:食管癌和贲门癌总切除率92．1%(174/189),其中根治性切除率为75．1%(142/189),探查率(未切除)为7．9%(15/189),三种术式的总吻合口瘘发生率为4．0%(7/174),无围手术期死亡。结论:三种术式可提高根治性切除率,大网膜包盖加固吻合口可减少瘘的发生率,食管胃分层吻合法可降低吻合口狭窄的发生率,临床治愈率高,围手术期死亡率低。     

犬获得性食管气管瘘模型建立及食管双瓣修补术实验研究

目的:1.建立犬食管气管瘘动物模型;2.观察食管壁双瓣修补气管缺损术局部组织病理学改变、愈合情况及该术式的治疗效果。方法:犬16只,随机分为2组,实验组(12只)建立犬食管气管瘘动物模型,行食管壁双瓣修补气管缺损术,对照组(4只)正常犬作为空白对照,术后支气管镜检查,术后1,2,8周处死动物,观察实验动物一般情况、修补局部的大体改变和组织病理学改变,测定气管狭窄指数。结果:成功建立了犬食管气管瘘动物模型;食管双瓣修补术后,实验组动物呛咳症状消失,均存活至预定时间,修补瓣和气管之间愈合良好,修补瓣血供良好,未出现气管狭窄情况,气管通畅度良好。结论:通过手术方式建立食管气管瘘模型方法可靠;食管双瓣修补术治疗TEF效果良好,值得进一步研究及推广。     

Water metabolism in the eel acclimated to sea water: from mouth to intestine

Eels seem to be a suitable model system for analysing regulatory mechanisms of drinking behavior in vertebrates, since most dipsogens and antidipsogens in mammals influence the drinking rate in the seawater eels similarly. The drinking behavior in fishes consists of swallowing alone, since they live in water and water is constantly held in the mouth for respiration. Therefore, contraction of the upper esophageal sphincter (UES) muscle limits the drinking rate in fishes. The UES of the eel was innervated by the glossopharyngeal-vagal motor complex (GVC) in the medulla oblongata (MO). The GVC neurons were immunoreactive to an antibody raised against choline acetyltransferase (ChAT), an acetylcholine (ACh) synthesizing enzyme, indicating that the eel UES muscle is controlled cholinergically by the GVC. The neuronal activity of the GVC was inhibited by adrenaline or dopamine, suggesting catecholaminergic innervation to the GVC. The AP and the commissural nucleus of Cajal (NCC) in the MO projected to the GVC and were immunoreactive to an antibody raised against tyrosine hydroxylase (TH), rate limiting enzyme to produce catecholamines from tyrosine. Therefore, it is likely that activation in the AP or the NCC may inhibit the GVC and thus relaxes the UES muscle, which allows for water to enter into the esophagus. During passing through the esophagus, the imbibed sea water (SW) was desalted to approximately 1/2 SW, which was further diluted in the stomach and arrived at the intestine as approximately 1/3 SW, almost isotonic to the plasma. Finally, from the diluted SW, the eel intestine absorbed water following the Na⁺–K⁺–2Cl⁻ cotransport (NKCC2) system. The NaCl and water absorption across the intestine was regulated by various factors, especially by peptides such as atrial natriuretic peptide (ANP) and somatostatin (SS-25 II). During desalination in the esophagus, however, excess salt enters into the blood circulation, which is liable to raise the plasma osmolarity. However, the eel heart was constricted powerfully by the hyperosmolarity, suggesting that the hyperosmolarity enhances the stroke volume to the gill, where excess salt was extruded powerfully via Na⁺–K⁺–2Cl⁻ cotransport (NKCC1) system.     

NADPH-diaphorase-positive nerve fibers associated with motor endplates in the rat esophagus: new evidence for co-innervation of striated muscle by enteric neurons

NADPH-diaphorase histochemistry was combined with demonstration of acetylcholinesterase and immunocytochemistry for calcitonin gene-related peptide to study esophageal innervation in the rat. Most of the myenteric neurons stained positively for NADPH-diaphorase, as did numerous varicose nerve fibers in the myenteric plexus, among striated muscle fibers, around arterial blood vessels, and in the muscularis mucosae. A majority of motor endplates (as demonstrated by acetylcholinesterase histochemistry or calcitonin gene-related peptide immunocytochemistry) were associated with fine varicose NADPH-diaphorase-positive nerve fibers. Analysis of brainstem nuclei, sensory vagal, spinal, and sympathetic ganglia in normal and neonatally capsaicin-treated rats, and comparison with anterogradely labeled vagal branchiomotor, preganglionic and sensory fibers led to the conclusion that NADPH-diaphorase-positive fibers on motor endplates originate in esophageal myenteric neurons. No association of NADPH-diaphorasepositive nerve fibers with motor endplates was found in other organs containing striated muscle. These results suggest extensive, presumably nitrergic, co-innervation of esophageal striated muscle fibers by enteric neurons. Thus, control of peristalsis in the esophagus of the rat may be more complex than hitherto assumed.     

Alterations to the subepithelial layers of the larval alimentary tract during metamorphosis in the sea lamprey Petromyzon marinus,with emphasis on tissue interactions

Light-microscopic histochemistry and conventional electron microscopy were used to study the changes to the subepithelial layers in the larval esophagus of the sea lamprey Petromyzon marinus during metamorphosis. During early stages of metamorphosis, smooth muscle cells of the muscularis mucosae and tunica muscularis dedifferentiate into myofibroblast-like cells, which make contact with the basal lamina of the overlying mucosal epithelium. During later stages, these myofibroblast-like cells redifferentiate into smooth muscle cells, reforming the muscularis mucosae and tunica muscularis. Alterations to the extracellular matrix occur concomitantly.     

Rat intestinal galactoside-binding lectin L-36 functions as a structural protein in the superficial squamous cells of the esophageal epithelium

Using an affinity purified antibody raised against the RI-H fragment of rat intestinal lectin L-36, the latter protein has been identified within the esophageal epithelium by means of ultracryotomy followed by immunogold labeling. The epithelium consists of 4 morphologically distinct cell-types, namely, the basal, spiny, granular and squamous cells, and each of these exhibits a different immunolabeling pattern. The basal cells form a layer on the basal lamina, and in these a diffuse cytoplasmic staining is observed. This basal cell layer is overlaid by spiny cells that extend many cell processes into wide intercellular spaces. In these cells, immunogold particles are found only on small granular inclusions consisting of an electron-lucent homogeneous substance. The granular cells from a third layer over the spiny cells, and are characterized by a number of large granular inclusions with an electron-dense core rimmed by a less electron-dense substance. Immunogold labeling is found on these granules, both on the core and peripheral region. Squamous cell-types constitute the most superficial layer of the epithelium. They are without granular inclusions, and immunogold labeling is confined to the cytoplasmic surface of the thickened plasma membrane. These findings suggest that L-36 is produced in the basal cells as free cytosolic protein, then becomes progressively aggregated into the granular inclusions of the spiny and granular cells, and is eventually transferred onto the cytoplasmic surface of the squamous cell plasma membrane where it may interact with complementary glycoconjugate(s) located at this site. The membrane lining substance thus formed may play a role in stabilizing the squamous cell membranes, thereby maintaining the structural integrity of the epithelium against mechanical stress coming from the esophageal lumen.