MUC-1的表达与肿瘤发生发展关系的研究进展

随着分子生物学的进展，近年来对肿瘤发生机制在分子水平上有了更新的认识，基因研究也越来越受到重视，MUC-1粘蛋白（mucin 1，MUC-1，CA15-3）是一种高度糖化的跨膜蛋白,在正常和恶性上皮细胞中被检测到，MUC-1基因在人类肿瘤的发生发展过程中起着重要作用，其与肿瘤侵袭和转移密切相关。血清MUC-1粘蛋白对肿瘤的辅助诊断、预后、临床监测有重要意义。本文就MUC-1在恶性肿瘤发生发展中的研究进展及其在临床中的应用作一综述。     

自身抗体在癌症早期检测中的应用

通过检测患者血清中自身抗体进行癌症诊断，与传统的肿瘤抗原的检测相比具有更高的敏感性和特异性。文章就血清自身抗体及在癌症早期诊断中的应用进行综述。     

自身抗体在癌症早期检测中的应用

通过检测患者血清中自身抗体进行癌症诊断，与传统的肿瘤抗原的检测相比具有更高的敏感性和特异性。文章就血清自身抗体及在癌症早期诊断中的应用进行综述。     

肿瘤相关抗原的抗体检测在原发性肝癌早期诊断中的价值

目的: 对10种肿瘤相关抗原(tumor-associated antigen,TAA)的自身抗体检测在原发性肝癌早期诊断中的价值进行评价,从而寻找一种真实可靠的肝癌早期诊断方法。 方法: 应用间接酶联免疫吸附试验(ELISA)检测原发性肝癌患者血清和正常人血清中的10种TAA(针对10个抗原Calnuc、CyclinE、CDK2、CIAP、RalA、p62、p53、Cy-clinB1、Koc、Imp1)的自身抗体,利用流行病学方法对检测结果的真实性进行评价。 结果: 每种TAA单独检测时,大多数指标的灵敏度偏低,但是肝癌患者抗体阳性率明显高于正常人;10种TAA两两联合起来时检测抗体阳性率,除了Calnuc抗体和CDK2抗体联合时阳性率为18.0%,其余联合均大于26.0%,明显高于单个抗体的检测结果,抗体阳性率最高达到50.0%,其中CyclinE和CIAP、CyclinE和Koc、CyclinE和Imp1等抗体阳性率均为50.0%;对10种TAA进行不同的组合,逐渐增加抗原数目进行检测,结果随着检测抗体的增多,诊断的灵敏度随之增加,10种抗体联合检测的灵敏度达到了88.0%,特异度也达到了86.2%,阳性预测值为84.6%,阴性预测值为89.3%。阳性似然比为6.25,阴性似然比为0.14,说明10种TAA检测肝癌的临床价值较高,Kappa值为0.74,提示该实验诊断结果与真实值之间高度一致。 结论: 利用10种TAA抗体组合检测肝癌具有较高的真实性,可作为现场高危人群筛检和临床中肝癌早期诊断的一种方法。     

血清中P53蛋白及其抗体的检测对肺癌诊断意义的研究

目的探讨血清中P53基因的表达产物P53蛋白和抗P53蛋白抗体的检测对肺癌患者诊断的应用价值。方法应用Westernblot方法．对28例肺癌患者和18例对照的血清中的P53蛋白和抗P53蛋白抗体进行检测。结果分别在7.14％（2／28）和14.29％（4／28）的肺癌患者的血清中检测到P53蛋白和抗P53蛋白抗体，而良性肺疾病和健康对照组血清中求检测到这两种蛋白抗体。结论检测患者血清中的P53蛋白和抗P53蛋白抗体对肺癌患者具有一定的诊断价值。     

检测MUC-1 mRNA对诊断非小细胞肺癌患者外周血微转移的意义

背景与目的:粘蛋白家族,包括MUC-1,通常被认为是上皮组织及其来源的肿瘤的产物.理论上如果在间叶组织来源的血液中找到MUC-1 mRNA即意味着癌转移.本研究探讨MUC-1 mRNA作为分子指标检测非小细胞肺癌患者外周血微转移的可行性.方法:应用巢式RT-PCR技术检测60例非小细胞肺癌患者外周血中MUC-1基因的表达,并以15例肺良性疾病患者及20名健康人外周血作为对照,同时使用相同的方法,扩增非上皮细胞来源的细胞株K562,HL-60及上皮细胞来源的细胞株A549、MCF-7,以观察靶基因的表达情况.结果:病患组MUC-1 mRNA表达阳性率达80.0%(48/60).15例肺良性疾病患者及20例健康人外周血中MUC-1 mRNA表达阳性率分别为60.0%(9/15),65.0%(13/20),K562、HL-60及A549、MCF-7细胞株经巢式RT-PCR也可以扩增出靶基因的条带.结论:MUC-1 mRNA不是检测非小细胞肺癌外周血微转移的可靠指标.     

血清 DKK1和 P53自身抗体联合检测对食管鳞状细胞癌的诊断价值

目的：探讨血清 DKK1（Dickkopf-1）和 P53自身抗体联合检测在食管鳞状细胞癌（ESCC）中的诊断意义。方法应用酶联免疫吸附试验检测126例 ESCC 患者和60例正常对照血清 DKK1和P53自身抗体的表达水平,采用受试者工作特征曲线（ROC）评价诊断效能。结果血清 DKK1水平和P53自身抗体在 ESCC 患者中的表达均明显高于正常对照[（673．09±343．82）pg／ml ∶（362．05±148．07）pg／ml,Z ＝6．158,P ＜0．0001;（0．398±0．546）∶（0．069±0．050）,Z ＝3．832,P ＜0．0001]。ROC 曲线显示,当血清 DKK1为最佳诊断临界值588．77 pg／ml 时,其在诊断 ESCC 的曲线下面积（AUC）为0．780（95％CI 为0．715～0．844,敏感性为61．9％,特异性为95．0％）。P53自身抗体诊断 ESCC 的AUC 为0．674（95％CI 为0．598～0．750,敏感性为45．3％,特异性为95．0％）。DKK1和 P53自身抗体联合检测诊断食管癌的 AUC 为0．843（95％CI 为0．788～0．897,敏感性为73．8％,特异性为95．0％）。在早期 ESCC,DKK1和 P53自身抗体联合检测具有更好的诊断效能,AUC 为0．903（95％CI 为0．845～0．961,敏感性为81．0％,特异性为95．0％）。结论血清 DKK1和 P53自身抗体可作为 ESCC 的血清诊断标志物,联合检测有助于 ESCC 的早期诊断。     

联合检测肿瘤相关抗原抗体在结直肠癌免疫诊断中价值的研究

目的:对多种肿瘤相关抗原 (tumor-associated antigen, TAA) 抗体的联合检测在结直肠癌(colorectal cancer , CRC)早期诊断中的价值进行评价,从而寻找一种真实可靠的CRC早期诊断方法.方法:采用ELISA法对46例CRC患者和58例健康人血清中的15种TAA抗体进行检测,并对检测结果行统计学分析后进行评价.结果:单独检测15种TAA时,大多数指标的敏感度都偏低(最高低于25%);剔除在单个抗体检测评价时没有统计学意义的3个抗体后,12种TAA两两组合后检测抗体阳性率最高可提高到41%,明显高于单个抗体的检测结果;对12种TAA进行不同的组合,逐渐增加抗原数目进行检测时,随着检测抗体的增多,诊断的敏感度随之增加,12种抗体联合检测的敏感度达到80.4%,特异度达82.8%,阳性预测值为78.7%,阴性预测值为84.2%.阳性似然比为5.81,阴性似然比为0.24,说明12种TAA检测CRC的临床价值较高,Kappa值为0.63,提示该实验诊断结果与真实值之间中度一致.结论:联合检测多种TAA抗体在CRC的早期诊断中具有一定的应用价值,从而为建立CRC早期体外非侵入性诊断技术的研究奠定了基础.     

血清半乳甘露聚糖检测对血液病患者侵袭性曲霉菌感染早期诊断的价值

 目的　评价血清半乳甘露聚糖（GM）检测对血液病患者侵袭性曲霉菌（IA）感染早期诊断的价值。方法　前瞻性采用酶联免疫吸附试验（ELISA）每周2次测定患者的GM水平,并计算该诊断试验的各项评价指标。结果　共有来自92例患者的113例次感染进入研究,检测血清标本472份,以确诊IA感染和临床诊断IA感染为真阳性组,排除IA感染为真阴性组,0.7为阈值,连续2次GM结果阳性为真阳性,该诊断试验的敏感度为83.3 %,特异度为91.1 %,阳性预测值为78.9 %,阴性预测值为93.1 %。GM阳性结果较痰培养阳性时间提前4 d（1～7 d）,比影像学改变提早7 d（1～14 d）,较抗真菌治疗提前6 d（1～15 d）。结论　通过ELISA方法进行血清GM检测可以快速、灵敏的为早期诊断IA感染提供有力证据。     

MUC-5AC核粘蛋白在早期胃癌中的表达和意义

目的探讨胃正常粘膜、不典型增生以及早期胃癌中MUC-5AC核粘蛋白的表达状态及临床病理意义.方法应用免疫组织化学s-p法检测人正常胃粘膜、不典型增生及早期胃癌组织中的表达.结果人正常胃粘膜中浅表1/3范围内广泛分布MUC-5AC核粘蛋白(100%).不典型增生和早期胃癌组织中表达的阳性率分别为70%和41.7%.早期胃癌中MUC-5AC核粘蛋白的表达与患者的性别、肿物的部位、大小、浸润深度、淋巴结转移和分化程度、异型度及癌旁肠化生和癌旁淋巴反应无关(P＞0.05).但与Laurem's分型(肠型、胃型、混合型)密切相关,且差异显著(P＜0.05).结论 MUC-5AC核粘蛋白在胃粘膜不典型增生和胃癌早期阶段即出现不同程度的异常表达,整个过程呈下调趋势.且与Lauren's分型密切相关.     

Monoclonal antibodies reacting with the MUC2 mucin core protein.

This study sought to produce monoclonal antibodies (MAbs) which reacted with the MUC2 core protein. Two MAbs [3A2 (IgG1) and 4F1 (IgM)] were produced by immunising female BALB/c mice with gel-formed mucin from the LS174T colon cancer cell line followed by a KLH conjugate of a 29 amino acid synthetic peptide whose sequence was derived from the variable number of tandem repeats (VNTR) region of a MUC2 cDNA clone. The MAbs reacted with synthetic MUC2 VNTR peptides but not synthetic MUC1 or MUC3 VNTR peptides, and showed specific reactivity in Western blotting with a high molecular weight protein produced by the LS174T colon carcinoma cell line. The use of shorter peptides indicated that the minimum peptide epitopes for these MAbs were different. Mab 3A2 reacted with amino acids 5-19 of the MUC2 VNTR by inhibition ELISA but not by direct ELISA, while 4F1 reacted with this peptide in both assays. Furthermore, 4F1 reacted in direct ELISA when a larger (29 amino acid) MUC2-derived peptide was coated onto the assay plate by incubating in carbonate buffer or by drying the peptide onto the assay plate, while 3A2 only reacted when this peptide was coated in carbonate buffer. The different specificity of the MAbs was also illustrated by the reactivity of 4F1 but not 3A2 with partially deglycosylated cystic fibrosis mucin. Immunohistochemical analysis with these MAbs revealed a strong reactivity with lung, gastric and colon tumours relative to normal tissue, with some breast and ovarian tumours also reacting. Both MAbs stained some normal goblet cells in the perinuclear region but not the mucin droplet or secreted mucin, indicating a reaction with immature (poorly glycosylated) mucin in the endoplasmic reticulum and/or golgi, but not with mature (fully glycosylated) mucin. In contrast, tumours showed strong diffuse cytoplasmic staining. 4F1 also showed weak apical cytoplasmic staining in some goblet cells and stained some tumours which showed no reactivity with 3A2. These antibodies should prove useful in the study of MUC2 structure and function, and in the diagnosis of some tumours.     

Tumor markers in breast cancer- European Group on Tumor Markers recommendations.

Recommendations are presented for the routine clinical use of serum and tissue-based markers in the diagnosis and management of patients with breast cancer. Their low sensitivity and specificity preclude the use of serum markers such as the MUC-1 mucin glycoproteins (CA 15.3, BR 27.29) and carcinoembryonic antigen in the diagnosis of early breast cancer. However, serial measurement of these markers can result in the early detection of recurrent disease as well as indicate the efficacy of therapy. Of the tissue-based markers, measurement of estrogen and progesterone receptors is mandatory in the selection of patients for treatment with hormone therapy, while HER-2 is essential in selecting patients with advanced breast cancer for treatment with Herceptin (trastuzumab). Urokinase plasminogen activator and plasminogen activator inhibitor 1 are recently validated prognostic markers for lymph node-negative breast cancer patients and thus may be of value in selecting node-negative patients that do not require adjuvant chemotherapy.     

Correlation Between Auto-antibodies to Survivin and MUC1 Variable Number Tandem Repeats in Colorectal Cancer

Aim: The aim of this study was to investigate the frequency and correlation between auto-antibodies tosurvivin and MUC1 variable number tandem repeats (VNTR) in colorectal cancer (CRC), which can providevaluable information for the design of immunotherapeutic vaccines for this disease. Methods: Enzyme-linkedimmunosorbent assays (ELISA) were used to examine the level of auto-antibodies against survivin and MUC1VNTR in the serum of 135 CRC patients and 95 healthy volunteers. Results: Using mean absorbance + 2 standarddeviations (SD) of the healthy samples as a cut-off value, the positive rates of survivin and MUC1 VNTR autoantibodiesin CRC were 31.1% and 18.5%, respectively. Altogether, the survivin and MUC1 VNTR positivesamples accounted for 36.3% of the CRC patients, and 7.4% were positive for both. Conclusion: A significantpositive correlation was found between levels of specific antibodies against survivin and MUC1 VNTR in theserum of CRC patients (r = 0.3652, P < 0.0001), suggesting that vaccines against both targets would elicit immuneresponses more effectively.     

Immunity to murine breast cancer cells modified to express MUC-1, a human breast cancer antigen, in transgenic mice tolerant to human MUC-1

The high incidence of breast cancer in women and the severity of the disease have stimulated a need for improved and novel forms of therapy. The product of the MUC-1 gene has been identified as a breast cancer-associated antigen in breast cancer patients. The gene has been cloned and sequenced. Transgenic mice were prepared that express human mucin and are naturally tolerant to the molecule, providing a unique opportunity to investigate immunotherapeutic strategies in experimental animals that might eventually be applied to breast cancer patients. A cell line (410.4) derived from a mouse mammary adenocarcinoma that arose in a BALB/c mouse was transduced with a retroviral vector (R1-MUC1-pEMSVscribe) that encoded MUC-1. After confirmation of the expression of human mucin, the cells (E3) were further modified by transduction with retroviral vectors encoding interleukin (IL)-2, IL-4, IL-12, or IFN-gamma to evaluate the effect of cytokine-secretion on the immunogenic properties of the cells in the MUC-1 transgenic mice. The results indicated that modification of the breast cancer cells to secrete IL-12 reduced and at times eliminated the tumorigenic growth properties of the cells. Under similar circumstances, progressively growing tumors formed in MUC-1 transgenic mice that received injections of unmodified E3 cells or with E3 cells modified to secrete IL-2, IL-4, or IFN-gamma. Immunity to breast cancer developed in MUC-1 transgenic mice that had rejected IL-12-secreting E3 cells because the animals were resistant to challenge with (non-cytokine-secreting) E3 cells. In vitro analyses confirmed the presence of T cell-mediated cytotoxicity toward the breast cancer cells in MUC-1 transgenic mice immunized with the IL-12-secreting cells. Our data obtained in a unique animal model system point toward an analogous form of therapy for breast cancer patients.     

Humoral immune responses of lung cancer patients against tumor antigen NY-ESO-1

The cancer-associated antigen NY-ESO-1 is expressed in a number of malignancies of different histological type. Patients with NY-ESO-1 expressing tumors have been shown to bear circulating autoantibodies against this antigen. In this study, we have assessed the NY-ESO-I autoantibody response in patients with lung cancer by a serum ELISA. Using a serum dilution of 1:400 we detected seroreactivity in 35 of 175 (20%) of patients. Incidence of autoantibodies was significantly higher in patients suffering from non small cell lung cancer (NSCLC, 23%) as compared to those with small cell lung cancer (SCLC, 9%). In the NSCLC group, NY-ESO-I antibody was significantly more frequent in patients with undifferentiated tumors (40%) as compared to patients with either adenocarcinoma or squamous cell carcinoma (15 and 29%). Our observations indicate that induction of NY-ESO-I autoantibodies depends on the histological subtype within a given tumor entity.     

Anti-tumor activity and immune responses induced by human cancer-associated mucin core peptide

Objective: To investigate the immune responses induced by apomucin which is a mixture of mucin core peptide, in mice for elucidating the role of mucin core peptide in the modulation of cancers. Methods: Apomucin was isolated from human pancreatic cancer cell line SW1990. The mice were immunized with this apomucin (10μg/time×6) plus DETOX. Results: When immunized, all mice developed delayed-type hypersensitivity (DTH) after challenged with apomucin or synthetic peptide MUC-2 or MUC-3, while the mice immunized with apomucin alone did not develop DTH. No antibodies were detected by ELISA after immunization. When the spleen cells of vaccinated mice were cocultured with this apomucin (10–50μg/ml) and rhIL-2(50U/ml)in vitro, the proliferated lymphocytes showed cytotoxicity against human cancer cells, including colon cancer, gastric cancer, pancreatic cancer and leukemia as measured by Cr-51 release assay. Antibodies against MUC-2 and MUC-3 could block the cytotoxicity. Conclusion: It was identified that a vaccine combined of apomucin and immune adjuvant DETOX can induce cellular immune response and anti-tumor cytotoxicity in mice. This study was supported by National Natural Science Foundation of China No. 39570792.     

Phagocytosis of breast cancer cells mediated by anti-MUC-1 monoclonal antibody, DF3, and its bispecific antibody

Human epithelial mucin, MUC-1, is commonly expressed in adenocarcinoma including 80% of breast cancers. erbB-2 is overexpressed in approximately 30% of breast cancers. Expression of MUC-1 and erbB-2 may be partially overlapping but discoordinate. Therefore, combined use of antibodies directed against these two antigens might increase the number of patients who benefit from immunotherapy. Monoclonal antibody (MAb) DF3 recognizes the MUC-1 tandem repeat. We investigated phagocytosis and cytolysis of cultured human breast cancer cells by monocyte-derived macrophages mediated by MAb DF3 and its bispecific antibody (BsAb) DF3xH22 with the second epitope directed against the Fc component of phagocytic cells. Purified monocytes from healthy donors were cultured with granulocyte macrophage colony-stimulating factor with or without IFN-gamma. antibody-dependent cellular phagocytosis (ADCP) and antibody-dependent cellular cytotoxicity (ADCC) assays were performed with these macrophages and MUC-1-expressing target cells (ZR75-1) in the presence of MAb DF3 and BsAb DF3xH22. ADCP was measured by two-color fluorescence flow cytometry using PKH2 (green fluorescent dye) and R-phytoerythrin (RPE) (red)-conjugated MAb against human CD14 and CD11b and was confirmed by confocal microscopy. ADCC was measured by (51)Cr release assay. Immunohistochemical staining studies of MUC-1 and erbB-2 were performed on 67 primary breast cancer tissues. Expression of MUC-1 and erbB-2 was partially overlapping but discoordinate in 67 consecutive breast cancers. Both MAb DF3 and BsAb DF3xH22 mediated ADCP. However, ADCP mediated by MAb DF3 was greater than that mediated by BsAb DF3xH22. ADCC as detected by (51)Cr release was not seen with either antibody. The addition of IFN-gamma to monocyte-derived macrophage cultures inhibited ADCP compared to granulocyte macrophage colony-stimulating factor alone. Given the partially overlapping but discoordinate expression of MUC-1 and erbB-2 in breast cancer, therapy directed toward both antigens should be considered. MAb DF3 and the BsAb DF3xH22, can effectively mediate phagocytosis of MUC-1-expressing target cells. Further investigations are needed to determine whether this antibody-induced phagocytosis results in long-term specific T-cell activation against MUC-1.