Serum neuron-specific enolase:A promising biomarker of silicosis

BACKGROUND Silicosis is a type of chronic pulmonary fibrosis caused by long-term inhalation of silica dust particles.There has been no ideal biomarker for the diagnosis and differential diagnosis of silicosis until now.Studies have found that elevated neuron-specific enolase(NSE)concentration in the serum of silicosis patients is helpful for diagnosis and severity assessment of the disease.However,the number of cases in these studies was not enough to arouse attention.AIM To investigate the clinical significance of serum NSE in the diagnosis and staging of silicosis.METHODS From January 2017 to June 2019,326 cases of silicosis confirmed in Quanzhou First Hospital Affiliated to Fujian Medical University were included in the silicosis group.A total of 328 healthy individuals or medical patients without silicosis were included in the control group.Serum NSE concentrations of all subjects were determined by electrochemical luminescence.RESULTS There were no significant differences in sex,age,smoking index and complications between the silicosis and control groups.The mean serum NSE concentration was 26.57±20.95 ng/mL in the silicosis group and 12.42±2.68 ng/mL in the control group.The difference between the two groups was significant(U=15187,P=0.000).Among the 326 patients with silicosis,103 had stage I silicosis,and the mean serum NSE concentration was 15.55±6.23 ng/mL.The mean serum NSE concentration was 21.85±12.05 ng/mL in 70 patients with stage II silicosis.The mean serum NSE concentration was 36.14±25.72 ng/mL in 153 patients with stage III silicosis.Kruskal-Wallis H test suggested that the difference in serum NSE concentration in silicosis patients in the three groups was significant(H=130.196,P=0.000).Receiver operating characteristic curve analysis indicated that the area under the curve was 0.858(95%confidence interval:0.828-0.888;P=0.000).When the NSE concentration was 15.82 ng/mL,the Jorden index was the largest,the sensitivity was 72%,and the specificity was 90%.CONCLUSION Serum NSE concentration may be a promising biomarker for the diagnosis and assessment of severity of silicosis.

Neuron-specific enolase expression in a rat model of radiation-induced brain injury following vascular endothelial growth factor-modified neural stem cell transplantation

BACKGROUND： Previous studies have shown that transplantation of vascular endothelial growth factor （VEGF）-modified neural stem cells （NSC） provides better outcomes, compared with neural stem cells, in the treatment of brain damage. OBJECTIVE： To compare the effects of VEGF-modified NSC transplantation and NSC transplantation on radiation-induced brain injury, and to determine neuron-specific enolase （NSE） expression in the brain. DESIGN, TIME, AND SETTING： The randomized, controlled study was performed at the Linbaixin Experimental Center, Second Affiliated Hospital, Sun Yat-sen University, China from November 2007 to October 2008. MATERIALS： VEGF-modified C17.2 NSCs were supplied by Harvard Medical School, USA. Streptavidin-biotin-peroxidase-complex kit （Boster, China） and 5, 6-carboxyfluorescein diacetate succinimidyl ester （Fluka, USA） were used in this study. METHODS： A total of 84 Sprague Dawley rats were randomly assigned to a blank control group （n = 20）, model group （n = 20）, NSC group （n = 20）, and a VEGF-modified NSC group （n = 24）. Rat models of radiation-induced brain injury were established in the model, NSC, and VEGF-modified NSC groups. At 1 week following model induction, 10 pL （5 ×10^4 cells/μL） VEGF-modified NSCs or NSCs were respectively infused into the striatum and cerebral cortex of rats from the VEGF-modified NSC and NSC groups. A total of 10μL saline was injected into rats from the blank control and model groups. MAIN OUTCOME MEASURES： NSE expression in the brain was detected by immunohistochemistry following VEGF-modified NSC transplantation. RESULTS： NSE expression was significantly decreased in the brains of radiation-induced brain injury rats （P 〈 0.05）. The number of NSE-positive neurons significantly increased in the NSC and VEGF-modified NSC groups, compared with the model group （P 〈 0.05）. NSE expression significantly increased in the VEGF-modified NSC group, compared with the NSC group, at 6 weeks following transplantation （P 〈 0.05）. CONCLUSION： VEGF-modified NSC transplantation increased NSE expression in rats with radiation-induced brain injury, and the outcomes were superior to NSC transplantation.

Flunarizine and lamotrigine prophylaxis effects on neuron-specific enolase, S-100, and brain-specific creatine kinase in a fetal rat model of hypoxic-ischemic brain damage

BACKGROUND： Calcium antagonists may act as neuroprotectants, diminishing the influx of calcium ions through voltage-sensitive calcium channels. When administered prophylactically, they display neuroprotective effects against hypoxic-ischemic brain damage in newborn rats. OBJECTIVE： To investigate the neuroprotective effects of flunarizine （FNZ）, lamotrigine （LTG） and the combination of both drugs, on hypoxic-ischemic brain damage in fetal rats. DESIGN AND SETTING： This randomized, complete block design was performed at the Department of Pediatrics, Shenzhen Fourth People＇s Hospital, Guangdong Medical College. MATERIALS： Forty pregnant Wistar rats, at gestational day 20, were selected for the experiment and were randomly divided into FNZ, LTG, FNZ ＋ LTG, and model groups, with 10 rats in each group. METHODS： Rats in the FNZ, LTG, and FNZ ＋ LTG groups received intragastric injections of FNZ （0.5 mg/kg/d）, LTG （10 mg/kg/d）, and FNZ （0.5 mg/kg/d） ＋ LTG （10 mg/kg/d）, respectively. Drugs were administered once a day for 3 days prior to induction of hypoxia-ischemia. Rats in the model group were not administered any drugs. Three hours after the final administration, eight pregnant rats from each group underwent model establishment hypoxia-ischemia brain damage to the fetal rats. Cesareans were performed at 6, 12, 24, and 48 hours later; and 5 fetal rats were removed from each mother and kept warm. Two fetuses without model establishment were removed by planned cesarean at the same time and served as controls. A total of 0.3 mL serum was collected from fetal rats at 6, 12, 24, and 48 hours, respectively, following birth. MAIN OUTCOME MEASURES： Serum protein concentrations of neuron-specific enolase and S-100 were measured by ELISA. Serum concentrations of brain-specific creatine kinase were measured using an electrogenerated chemiluminescence method. RESULTS： Serum concentrations of neuron-specific enolase, S-100, and brain-specific creatine kinase were significantly higher in the hypoxic-ischemic fetal rats, compared with the non-hypoxic-ischemic group. Serum concentrations of neuron-specific enolase, S-100, and brain-specific creatine kinase were significantly less in the FNZ, LTG, and FNZ ＋ LTG groups following ischemia, compared with the model group （P 〈 0.01）. However, these values were significantly greater in the FNZ and LTG groups, compared with the FNZ ＋ LTG group, following ischemia （P 〈 0.01）. CONCLUSION： Preventive antenatal use of oral FNZ and LTG has positive neuroprotective effects on intrauterine hypoxic-ischemic brain damage. The combined effect of these two drugs is superior.

Electrophoretic Determination of Aqueous and Serum Neuron-specific Enolase in the Diagnosis of Retinoblastoma

Purpose: Neuron-specific enolase (NSE) of containing γ-enolase is considered valuable in the diagnosis of tumours of neuroectodermal origin.Method : We used rapid electrophoretic method on cellulose acetate plate to determine the pattern of enolase isoenzymes in 21 aqueous humor and 23 serum specimens from retinoblastoma (Rb) and 21 aqueous and 25 serum specimens from 25 control cases to evaluate NSE in the diagnosis of Rb. The assay allowed assessment of all three major isoenzymes (aa,aγ and γγ),and NSE relative activity and its percentage in the total relative activity of the three enolase isoenzymes were assessed by means of fluorometer.Result: Aqueous from all patients with Rb contained aa,ar and rr isoenzymes and presented strong postitive, the positive rate of NSE being 100% and its relative activity accounting for 45 ± 9% of the total relative activity of the 3 enolase isoenzymes; No enolase was detectable in control aqueous with cataract, glaucoma and Coats's diseases (4 cases),but in two

NEURON-SPECIFIC ENOLASE IN PATIENTS WITH ACUTE ISCHEMIC STROKE AND RELATED DEMENTIA

Neuron-specific enolase (NSE) levels of cerebrospinal fluid (CSF) were measured in 39 patients with ischemic stroke and 15 controls. There was a significant increase of CSF NSE in acute ischemic stroke patients as compared with the controls. The altered CSF NSE levels correlated well with the infarct size in CT scan. The CSF NSE levels were higher in 6-multiinfarct dementia (MID) patients who were diagnosed after 6-month follow-up than those in 22 non-MID patients of this series. Our research supports the view that CSF NSE can be a useful biochemical marker for brain ischemia. The importance of CSF NSE in the study of dementia related to ischemic stroke is worth further studies.

Choice of serum tumor markers in patients with small cell lung cancer:progastrin-releasing peptide,neuron-specific enolase,and carcinoembryonic antigen

Lung cancer is a leading cause of cancer-related deaths worldwide.It mainly consists of 2 histological types:small cell lung cancer(SCLC)and non-small cell lung cancer(NSCLC,including squamous cell carcinoma and adenocarcinoma).The present study aimed to assess the role of serum progastrin-releasing peptide(ProGRP),neuron-specific enolase(NSE),and carcinoembryonic antigen(CEA)and their combinations in the histological diagnosis of lung cancer(specially SCLC),which is of great importance for the initiation of treatment and prognostic implications.Serum ProGRP,NSE,and CEA were determined by the electrochemiluminescence immunoassay(ECLIA)in 66 patients with SCLC,73 with adenocarcinoma,44 with squamous cell carcinoma,45 with non-malignant pulmonary diseases,and 50 healthy controls.Receiver operating characteristic curves were constructed to compare the predictive ability of each biochemical marker and their combined detection models to discriminate among the patients with lung cancers of different histological groups,benign pulmonary diseases and healthy individuals.In the ECLIA detection system,ProGRP showed the sensitivity and specificity for SCLC diagnosis were 71.2%and 91.1%to 93.2%,respectively.Among the markers,the largest area under the ROCs was for ProGRP in discriminating SCLC from benign pulmonary diseases,squamous cell carcinoma and adenocarcinoma(0.815,0.859,and 0.835,respectively),which indicated that ProGRP was the most efficient marker for identifying SCLC.Besides,ProGRP and NSE exhibited almost equivalent diagnostic performance in discriminating SCLC from benign diseases.As for squamous cell carcinoma,we recommended proGRP,while for adenocarcinoma,the combination of proGRP and CEA was preferred.Remarkably,when ProGRP≤66pg/mL,CEA was of great value in diagnosing SCLC and adenocarcinoma.If CEA≤5ng/mL,the patient was at higher risk for SCLC,whereas the patient was more likely to be diagnosed with adenocarcinoma.Our study provided promising information about the diagnostic values of serum ProGRP,NSE,CEA in distinguishing SCLC from benign pulmonary diseases and NSCLC,which was of crucial clinical significance in the early diagnosis and therapy of SCLC.

Use of neuron-specific enolase to predict mild brain injury in motorcycle crash patients with maxillofacial fractures: A pilot study

Purpose: Mild traumatic brain injury (TBI) is common but accurate diagnosis and its clinical consequences have been a problem. Maxillofacial trauma does have an association with TBI. Neuron-specific enolase (NSE) has been developed to evaluate neuronl damage. The objective of this study was to investigate the accuracy of NSE serum levels to detect mild brain injury of patients with sustained maxillofacial fractures during motor vehicle accidents. Methods: Blood samples were drawn from 40 healthy people (control group) and 48 trauma patients who has sustained isolated maxillofacial fractures and mild brain injury in motor vehicle accidents. Brain injuries were graded by Glasgow Coma Scale. In the trauma group, correlations between the NSE serum value and different facial fracture sites were also assessed. Results: The NSE serum level (mean ± SD, ng/ml) in the 48 patients with maxillofacial fractures and mild TBI was 13.12 ± 9.68, significantly higher than that measured in the healthy control group (7.72 ± 1.82, p < 0.001). The mean NSE serum level (ng/ml) in the lower part of the facial skeleton (15.44 with SD 15.34) was higher than that in the upper facial part (12.42 with SD 7.68);and the mean NSE level (ng/ml) in the middle-and lower part (11.97 with SD 5.63) was higher than in the middle part (7.88 with SD 2.64). Conclusion: An increase in NSE serum levels can be observed in patients sustained maxillofacial fractures and mild brain injury.

Significance of serum neuron-specific enolase in patients with acute traumatic brain iniurv

Objective: To study the association between serum neuron-specific enolase (NSE) and the extent of brain damage and the outcome after acute traumatic brain injury (TBI). Methods: The release patterns of serum NSE in 78 patients after acute TBI were analyzed by using the enzyme linked immunosobent assay. The levels of NSE were compared with Glasgow coma scale, the category of brain injury and the outcome after 6 months of injury. Results: There were different NSE values in patients with minor (12.96 μg/L±2.39 μg/L), moderate (23.44 μg/L±5.33 μg/L) and severe brain injury (42.68 μg/L±4.57 μg/L). After severe TBI, the concentration of NSE in patients with epidural hematomas was 13.38 μg/L±4.01 μg/L, 24.03 μg/L±2.85 μg/L in brain contusion without surgical intervention group, 55.20 μg/L±6.35 μg/L in brain contusion with surgical intervention group, and 83.85 μg/L±15.82 μg/L in diffuse brain swelling group. There were close correlations between NSE values and Glasgow coma scale (r=-0.608, P<0.01) and the extent of brain injury (r=0.75, P<0.01). Patients with poor outcome had significantly higher initial and peak NSE values than those with good outcome (66.40 μg/L±9.46 μg/L, 94.24 μg/L±13.75 μg/L vs 32.16 μg/L±4.21 μg/L, 34.08 μg/L±4.40 μg/L, P<0.01, respectively). Initial NSE values were negatively related to the outcome (r=-0.501, P<0.01). Most patients with poor outcomes had persisting or secondary elevated NSE values. Conclusions: Serum NSE is one of the valuable neurobiochemical markers for assessment of the severity of brain injury and outcome prediction.

鼻咽癌高癌家系气虚质癌患者α-enolase的表达活性

目的探讨高癌家系气虚质鼻咽癌初诊患癌组织中α-enolase的表达活性水平及其临床病理意义。方法分别收集高癌家系中气虚质鼻咽癌初诊患者、鼻咽黏膜慢性炎症患者各9例,非高癌家系气虚质鼻咽癌初诊患者12例的鼻咽黏膜组织,Real Time PCR分别检测ENO1 mRNA表达活性,Western blotting检测ENO1蛋白表达水平,比较分析其组间差异及其临床病理意义。结果高癌家系组、非高癌家系气虚质鼻咽癌患者组、健康人鼻咽组织中ENO1 mRNA的△Ct分别为2.45±0.42,3.47±0.28,4.49±0.51;2^(-△△Ct)值分别为4.09±1.27,1.97±0.38,1.00±0.30;各组ENO1蛋白相对表达量依次为2.94±0.81、1.73±0.53、1.27±0.25;ENO1 mRNA及其蛋白表达水平组间差异均具有统计学意义(P<0.05)。结论高癌家系气虚质初诊鼻咽癌患者ENO1 mRNA及蛋白表达水平的明显上调具有显著的临床病理意义,对高癌家系气虚质鼻咽癌患者早期筛查及诊断可能具有较好的临床应用价值。

Regulation of enolase activation to promote neural protection and regeneration in spinal cord injury

Spinal cord injury(SCI)is a debilitating condition characterized by damage to the spinal cord resulting in loss of function,mobility,and sensation with no U.S.Food and Drug Administration-approved cure.Enolase,a multifunctional glycolytic enzyme upregulated after SCI,promotes pro-and anti-inflammatory events and regulates functional recovery in SCI.Enolase is normally expressed in the cytosol,but the expression is upregulated at the cell surface following cellular injury,promoting glial cell activation and signal transduction pathway activation.SCI-induced microglia activation triggers pro-inflammatory mediators at the injury site,activating other immune cells and metabolic events,i.e.,Rho-associated kinase,contributing to the neuroinflammation found in SCI.Enolase surface expression also activates cathepsin X,resulting in cleavage of the C-terminal end of neuron-specific enolase(NSE)and non-neuronal enolase(NNE).Fully functional enolase is necessary as NSE/NNE C-terminal proteins activate many neurotrophic processes,i.e.,the plasminogen activation system,phosphatidylinositol-4,5-bisphosphate 3-kinase/protein kinase B,and mitogen-activated protein kinase/extracellular signal-regulated kinase.Studies here suggest an enolase inhibitor,ENOblock,attenuates the activation of Rho-associated kinase,which may decrease glial cell activation and promote functional recovery following SCI.Also,ENOblock inhibits cathepsin X,which may help prevent the cleavage of the neurotrophic C-terminal protein allowing full plasminogen activation and phosphatidylinositol-4,5-bisphosphate 3-kinase/mitogen-activated protein kinase activity.The combined NSE/cathepsin X inhibition may serve as a potential therapeutic strategy for preventing neuroinflammation/degeneration and promoting neural cell regeneration and recovery following SCI.The role of cell membrane-expressed enolase and associated metabolic events should be investigated to determine if the same strategies can be applied to other neurodegenerative diseases.Hence,this review discusses the importance of enolase activation and inhibition as a potential therapeutic target following SCI to promote neuronal survival and regeneration.

微小牛蜱Enolase基因在昆虫细胞中的表达及鉴定

为对微小牛蜱烯醇酶(Enolase)基因进行真核表达及免疫反应原性分析,本研究采用RT-PCR方法扩增微小牛蜱Enolase基因的开放阅读框(ORF)序列,将其克隆至pFastBac1载体中构建重组质粒pFastBac1-Enolase,随后转化至大肠杆菌DH10Bac细胞中制备了重组杆粒Bacmid-Enolase,转染至SF9细胞以获得重组Enolase蛋白。通过双酶切、PCR及测序鉴定显示Enolase基因正确插入。利用SDS-PAGE分析表达产物,结果显示目的蛋白大小约为48 ku。利用western blot对重组蛋白进行分析,结果表明目的蛋白能被微小牛蜱抗原免疫兔阳性血清识别,具有良好的免疫反应原性。凝血酶时间(TT)的测定结果显示,Enolase重组蛋白能显著延长TT,具有抗凝血活性。本研究首次采用Bac-to-Bac杆状病毒表达系统对微小牛蜱Enolase基因进行真核表达,并鉴定了重组蛋白的免疫反应原性,为开展Enolase免疫原性分析及功能研究奠定了基础。

宫颈癌中α-enolase表达及其与HPV感染的关系

目的探讨烯醇化酶-α(α-enolase)蛋白在宫颈鳞癌中的表达及其与HPV感染的关系。方法应用免疫组化PV-9000两步法检测30例慢性宫颈炎、61例宫颈上皮内瘤变(cervical intraepithelial neoplasia,CIN)和70例宫颈鳞癌组织中α-enolase蛋白的表达,同时应用基因芯片技术检测70例宫颈鳞癌中HPV感染情况。结果 (1)α-enolase蛋白表达于细胞质和(或)胞核中。在慢性宫颈炎、CIN和宫颈癌中,α-enolase蛋白在胞质表达的阳性率分别为4.17%(1/24)、18.5%(10/54)和54.3%(38/70),表达依次增强(P=0.000)。(2)宫颈癌中HPV总感染率为97.1%(68/70),共检出8种HPV基因型,分别为HPV16、18、58、31、52、59、66、68,构成比为80.0%、14.3%、4.3%、4.3%、2.9%、2.9%、1.43%、1.43%。双重感染10例,占14.3%。HPV16、18为主要致病基因型。(3)宫颈癌中α-enolase蛋白表达的定位与HPV16/18感染呈正相关(r=0.340,P=0.012)。结论宫颈鳞癌中α-enolase蛋白表达与HPV16/18感染密切相关,二者在宫颈鳞癌的发生过程中可能起着协同作用。

enolase-α蛋白在鼻咽癌组织中的表达及意义

目的探讨烯醇化酶α(enolase-α)在人鼻咽癌(NPC)组织中的表达及其与NPC发生发展的关系。方法采用组织微阵列技术结合免疫组化法检测18例正常鼻咽黏膜组织、24例癌旁鼻咽上皮组织和90例NPC组织标本中enolase-α的表达,并分析其与临床参数的关系。结果enolase-α在正常鼻咽黏膜上皮、癌旁鼻咽上皮和NPC组织中的表达分别为94.4%、91.7%和52.2%,NPC组织表达enolase-α低于正常鼻咽黏膜上皮和癌旁鼻咽上皮(P<0.01)。enolase-α在NPC组织细胞核的表达水平低于正常鼻咽黏膜上皮和癌旁鼻咽上皮组织(P<0.01)。NPC患者中enolase-α表达与患者性别、组织学分类、淋巴结转移及临床TNM分期无关(P>0.05)。结论enolase-α蛋白表达下调与NPC的发生可能相关。

α-enolase蛋白在口腔鳞状细胞癌组织中的表达及其临床意义

目的探讨α-enolase蛋白在口腔鳞状细胞癌组织中的表达及其与口腔鳞癌发生发展的关系。方法采用ELISA法对40例口腔鳞癌组织、10例口腔正常粘膜组织中α-enolase蛋白的表达情况进行检测。结果 1.口腔鳞状细胞癌组织中α-enolase蛋白表达明显高于正常粘膜组织(P<0.01)。2.α-enolase蛋白在低、中、高分化口腔鳞状细胞癌组织中的表达均有差异(P<0.05),差异具有统计学意义,高分化鳞癌的α-enolase浓度>中分化鳞癌>低分化鳞癌;3.有淋巴结转移、TNM分期为III+IV期者α-enolase的表达分别明显高于无淋巴结转移、TNM分期为I+II期者(P<0.05)。4.α-enolase在口腔鳞状细胞癌中的表达与淋巴结转移、TNM分期、肿瘤分化程度密切相关(P<0.05),而与患者的年龄、性别无关(P>0.05)。结论口腔鳞状细胞癌组织中α-enolase蛋白表达上调,α-enolase可能在口腔鳞状细胞癌的发生、发展中起重要作用。

STAT-3与Enolase-1在乳腺癌组织中的表达及意义

目的探讨乳腺癌组织中信号转导及转录活化因子3(STAT-3)和糖酵解酶烯醇化酶(Enolase-1)的表达与雌孕激素受体、淋巴结转移、临床分期、病理分级等的关系。方法免疫组织化学SP法检测126例乳腺癌和26例乳腺良性病变组织中STAT-3与Enolase-1的表达情况,并分析他们与临床病理参数之间的关系。结果乳腺癌组织中STAT-3和Enolase-1的阳性表达率分别为82.5%和77.8%,明显高于乳腺良性病变组织(11.5%和7.7%),差异均有统计学意义(χ2=42.416,P<0.05;χ2=57.211,P<0.05);在有淋巴结转移的乳腺癌组织中阳性表达率分别为85.7%和90.5%,高于无淋巴结转移组,差异均有统计学意义(χ2=9.184,P<0.05;χ2=11.014,P<0.05)。相关分析表明STAT-3和Enolase-1的表达呈正相关。在雌激素受体阳性(ER+)和/或孕激素受体阳性(PR+)或表皮生长因子受体2阳性(HER-2+)乳腺癌组织中,有淋巴结转移组中STAT-3和Enolase-1的表达水平均明显高于无淋巴结转移组,差异均有统计学意义(P<0.05)。结论 STAT-3与Enolase-1的表达与乳腺癌的发生发展及侵袭转移相关,且二者有协同作用(r=0.379,P<0.05)。在ER+和(或)PR+或HER-2+乳腺癌组织中,STAT-3和Enolase-1的高表达与淋巴结转移相关。在高表达STAT-3的乳腺癌组织中Enolase-1表达也较高,其联合检测可作为判断乳腺癌恶性程度、评价预后及指导治疗的一项评估指标。

凡纳滨对虾烯醇酶基因Enolase的克隆及表达分析

以生长性状分离的凡纳滨对虾家系的肌肉组织,构建凡纳滨对虾生长性状差减cDNA文库,斑点杂交筛选获得Enolase基因,克隆获得全长编码区序列。序列分析表明,序列包含1 305 bp的开放阅读框,推测编码的蛋白质含434个氨基酸,预测的分子量为47.4 ku,等电点为5.67。通过荧光定量PCR法研究了Enolase基因在凡纳滨对虾的不同组织和不同生长期的肌肉组织中的表达情况,结果显示,Enolase基因在凡纳滨对虾的心脏、肝胰腺、肌肉、胃、肠、眼柄、鳃和血等组织或器官中都有表达;在肌肉组织中表达量最高,在肝胰腺中表达量最低;在日龄为125 d的对虾肌肉组织中表达量最高,在日龄为80 d的对虾肌肉组织中表达量最低。

α-enolase在宫颈癌中的表达及其意义

目的探讨α-烯醇化酶(enolase)在宫颈癌组织中的表达及其意义。方法应用免疫组织化学方法检测30例慢性宫颈炎、61例宫颈上皮内瘤样病变(CIN)和99例宫颈癌组织中α-enolase的表达。结果α-enolase蛋白在慢性宫颈炎、CIN和宫颈癌中阳性表达逐渐增强(P=0.000);蛋白定位则由胞质胞核逐渐转移至胞质(P=0.000)。宫颈癌中,α-enolase蛋白表达与患者年龄、肿瘤分级、淋巴结转移无关(P>0.05),与组织学分型有关(P=0.020);α-enolase蛋白定位与患者年龄、有无淋巴结转移及组织学分型无关(P>0.05),与肿瘤分级有关。α-enolase蛋白随肿瘤级别的增加而趋向出现在胞质中(P=0.019)。结论α-enolase蛋白的异常表达可能与宫颈癌的发生有关,有望成为宫颈组织早期癌变的有用标记物。

表面蛋白烯醇化酶Enolase在S.suis 2感染中的角色

目的通过克隆表达,在获得具有酶活性的猪链球菌2型(S.suis2)05ZYH33重组烯醇化酶Enolase蛋白的基础上,旨在继续探索其在细菌粘附和引发免疫下调作用中的角色。方法流式细胞术(FCM)、Hep2细胞粘附竞争试验、免疫空斑试验。结果流式细胞术FCM的细胞定位显示Enolase可以部分存在S.suis205ZYH33细菌的表面;Hep2细胞粘附竞争试验表明猪链球菌表面Enolase参与细菌对宿主细胞的黏附作用;免疫空斑实验的结果揭示Enolase在抑制宿主特异性免疫应答中发挥作用。结论 Enolase作为一个表面蛋白,确实在S.suis2感染中扮演一定的角色。

α-enolase在大肠癌与癌旁组织中的表达及意义

目的观察α-enolase在大肠癌中的表达情况及意义,探讨其与大肠癌发生发展及转移的关系。方法采用液质联用技术对18例大肠癌患者的癌组织和癌旁组织进行蛋白质谱的检测、鉴定和分析,并通过免疫蛋白印迹实验进一步确证。结果α-enolase在大肠癌组织中表达高于癌旁正常组织。结论高表达的α-enolase可能与大肠癌的发生发展有关。

人ENO1(Enolase-α)基因的原核表达、蛋白纯化及多克隆抗体的制备

目的:构建人ENO1(human Enolase-α)基因的原核表达质粒,诱导重组蛋白的表达,利用纯化后的蛋白制备具有活性的ENO1多克隆抗体。方法:用PCR方法从胃癌MKN45细胞全基因组中扩增出目的基因ENO1,将目的基因和原核表达载体pET30a(+)分别双酶切,回收酶切产物并用T4连接酶连接,获得重组质粒pET30a(+)-ENO1。将重组质粒转入大肠埃希菌菌株BL21,经异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,用Ni-NTA亲和层析柱纯化,利用纯化的ENO1活性蛋白作为抗原免疫家兔,制备抗血清多克隆抗体,并用Western blot检测其特异性。结果:目的基因与原核表达载体经双酶切鉴定所切下的片段大小与理论值相符,测序结果表明重组人ENO1基因序列与NCBI(NM_001428.3)公布序列一致。经SDS-PAGE分析,结果显示重组蛋白的分子量约在47kDa,条带单一,无杂带出现,主要以可溶性形式存在。Western blot证实多克隆抗体制备成功。结论:成功表达、纯化了ENO1融合蛋白,制备了具有高特异性的多克隆抗体,为进一步肿瘤疫苗的研制和肿瘤标志物快速诊断的研究奠定基础。