CEA和MUC-1mRNA在大肠癌中的表达及临床意义

[目的]探讨大肠癌患者外周血中CEA、MUC-1 m RNA的表达及临床意义。[方法]应用巢式RT-PCR技术检测20名健康人、20例大肠腺瘤疾病患者和90例大肠癌患者外周血CEA m RNA和MUC-1 m RNA的表达。[结果]大肠癌患者CEA m RNA和MUC-1 m RNA表达的阳性率分别为53.3%(48/90)和52.2%(47/90);健康人外周血中无CEA m RNA,有12名MUC-1 m RNA表达,表达率为60.0%(12/20);大肠腺瘤疾病患者无CEA m RNA表达,有10例MUC-1 m RNA表达,表达率为50.0%(10/20)。大肠癌患者中CEA、MUC-1 m RNA表达与肿瘤分期、淋巴结转移有关(P<0.05)。[结论]CEA作为检测大肠癌患者外周血微转移的分子标志物具有良好的灵敏度和特异性,同时检测CEA和MUC-1可作为判断大肠癌病情进展的指标。     

Experimental studies of tumor radioimmunodetection using antibody mixtures against carcinoembryonic antigen (CEA) and colon-specific antigen-p (CSAp)

Experiments with the GW-39 human colonic carcinoma growing in hamsters showed that injection of radioactive antibody to a colorectal-specific, tumor-associated antigen, CSAp, results in better tumor radiolocalization than was seen previously with radioantibodies to carcinoembryonic antigen (CEA). However, a mixture of both radioactive antibodies resulted in potentiation of CEA-tumor radioimmunodetection without affecting CSAp-tumor radiolocalization. Hence, multi-marker antibody mixtures may be the method of choice in cancer radioimmunodetection.     

Epidermal growth factor receptor, MUC-1 and MUC-2 in bladder cancer

The aim of this study was to investigate the immunohistochemical expression of epidermal growth factor receptor (EGFR), mucin-1 (MUC-1) and mucin-2 (MUC-2) proteins in primary bladder carcinomas and to compare EGFR and MUC staining patterns with the histological findings, grade and stage of bladder carcinoma. Fifty-six surgical specimens obtained from superficial and deeply invasive bladder carcinomas were studied. Of the 56 bladder tumors 42 (75%) expressed EGFR, 34 (60.71%) MUC-1 and 15 (26.78%) MUC-2; while 7 tumors (12.5%) coexpressed MUC-1 and MUC-2 proteins. Immunohistochemical scores showed higher levels of EGFR than of MUC-1 (P <0.05) and MUC-2 (P = 0.000) and higher levels of MUC-1 than MUC-2 (P = 0.0010). EGFR and MUC-1 expression was stronger in high-grade tumors (grade 2/3) than in low-grade (grade 1/2) ones (P <0.05) and stronger in muscle invasive tumors (T2-T4) than in superficial (Ta-T1) ones. Linear regression showed a significant (P <0.05) correlation between EGFR and MUC-1 proteins, but no correlation between EGFR and MUC-2 or between MUC-1 and MUC-2. Immunohistochemical expression of EGFR, MUC-1 and MUC-2 increases as primary bladder carcinomas acquire a more aggressive phenotype. Differences in the distribution of EGFR and mucins within the urothelium may be of diagnostic and prognostic value. These antigens may be useful as markers for bladder malignancy.     

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。     

Relationship among CEA expression in tumor,CEA serum level and radioimmunoguided surgery in colorectal cancer

Current chemotherapeutic agents offer <20% of survival benefits, and surgery still plays an important role in treating colorectal cancer[1]. Although more than two-thirds of patients are candidates for curative surgery, local recurrence occurs in as many as 40% after curative resection[2]. Consequently, a diagnostic tool to enable accurate tumor localization is urgently needed to allow precise and complete removal. Radioimmunoguided surgery (RIGS), originally designed by Aitken, et al.[3], i…     

Prognostic significance of MUC-1 expression in systemic anaplastic large cell lymphoma.

Systemic anaplastic large cell lymphoma (ALCL) frequently carries the t(2;5)(p23;q35) and overexpresses anaplastic lymphoma kinase (ALK). MUC-1, a highly glycosylated transmembrane protein, is detected in normal and malignant epithelial cells and has been associated with a poorer patient survival in various human malignancies. We have shown previously that MUC-1 is expressed as a consequence of t(1;14)(q21;32) in a subset of diffuse large B-cell lymphomas. ALCLs are known to express MUC-1, but its clinical significance is undefined. For this study, eligible patients with ALCL were HIV negative, received anthracycline-containing regimens, and had pretreatment archival tissue. Expression of MUC-1 and ALK was determined immunohistochemically after heat-induced antigen retrieval. A 10% cutoff for MUC-1 positivity was used. We identified 63 patients with systemic ALCL (22 ALK+, 41 ALK-) with a median age of 47 years, and 41 were male. MUC-1 was detected in 16 of 22 (73%) ALK-positive and 20 of 41 (49%) ALK-negative ALCL (P = 0.06, chi(2) test). MUC-1 expression was not associated with apoptotic rate as detected by terminal deoxynucleotidyl transferase-mediated nick end labeling assay or proliferation index as evaluated by MIB-1 antibody. For 48 patients with ALCL (16 ALK+, 32 ALK-) and complete clinical follow-up, 5-year progression-free survival (PFS) was 39.7% for patients with MUC-1-positive tumors versus 75.2% (P = 0.027 by Log-rank) for patients with MUC-1-negative tumors. For the ALK-negative ALCL group of 32 patients, the 5-year PFS was 26 versus 70.8% for patients with MUC-1-positive versus MUC-1-negative tumors (P = 0.0096 by Log-rank). For the ALK-positive ALCL group of 16 patients, the 5-year PFS was 52 versus 100% for patients with MUC-1-positive versus MUC-1-negative tumors (P, not significant). In summary, MUC-1 is frequently expressed in systemic ALCL, and its expression is associated with significantly inferior outcome in patients untreated previously with ALK-negative tumors. Future studies should explore the underlying molecular mechanisms of MUC-1 expression in these tumors and its role as a target for novel therapeutic strategies.     

Combination vascular targeted and tumor targeted radioimmunotherapy

Rat MAb 201B, which binds to murine thrombomodulin, can deliver up to 50% of the injected dose of attached radioisotopes to the lung vascular endothelium. We have shown previously that intravenous injection of about 30 microCi of 213Bi-MAb 201B, which delivers about 15 Gy of alpha irradiation to the lung, is capable of eradicating small lung colonies (500-1000 cells) of the mammary tumor line, EMT-6. Larger tumors (> 5000 cells) were not completely cured by this vascular targeted radioimmunotherapy (VT-RAIT) approach. We reasoned that VT-RAIT might make the lung vessels serving the tumor cells more permeable, allowing MAb targeted to the tumor cells to extravasate more readily and mediate more efficient standard radioimmunotherapy (RAIT). Distribution experiments with the tumor targeted MAb 13A (RAIT MAb), following VT-RAIT, did not demonstrate a large increase in tumor uptake; however, microautoradiography did indicate that MAb 13A was distributed more evenly throughout the tumor when administered after VT-RAIT. Therapy experiments on lung tumors of approximately 5000 cells each, combining 213Bi-MAb 201B (VT-RAIT) with 213Bi-MAb 13A (RAIT) 24 hours later, resulted in a better outcome (3 cured/10 at risk) than for control groups: RAIT only (0/10), VT-RAIT only (1/10), or no therapy (0/10). RAIT therapy delivered 48 hours after VT-RAIT had no apparent benefit. 213Bi-MAb 201B VT-RAIT followed by 90Y-MAb 13A Fab' RAIT showed only a slight improvement in tumor cures (2/10) over that in control groups: (0/9), (0/10), (0/10), respectively. These results suggest that optimal timing, dosage, and choice of MAb for RAIT should enhance the double MAb therapy approach significantly.     

Reoxygenation in the RIF-1 tumor after chemotherapy.

The proportion of hypoxic cells in the RIF-1 tumor was observed for 24 hr after treatments with bleomycin, cisplatin, cyclophosphamide, and mitomycin C. The assays were based upon the paired survival curve method for determining the hypoxic fraction using irradiation of aerobic and artificially hypoxic tumors. It was observed that at 1/2 hr after bleomycin, hypoxic fraction was elevated but returned to baseline levels by 2 hr. Following cisplatin, hypoxic fraction did not rise above baseline. However, after cyclophosphamide, hypoxic fraction was elevated and did not return to baseline over the 24-hr observation period of this study. At 1/2 hr after mitomycin treatment, the hypoxic fraction was raised but within 1 hr returned to baseline. These results indicate that tumor reoxygenation varies after different drug treatments and that the determination of drug specificity for aerobic versus hypoxic cells may be strongly biased by the choice of the time after treatment for making the determination.     

循环肿瘤细胞数目与血清中Cyfra21-1、CEA、Fg表达水平的关系

目的:探讨肿瘤患者外周血循环肿瘤细胞(CTCs)与常用的肿瘤指标细胞角质素19可溶片段（Cyfra21-1）、癌胚抗原（CEA）及凝血功能指标纤维蛋白原（Fg）表达水平的关系,比较CTCs阳性组和阴性组转移灶数目差异。方法:采集66例进展期晚期肺癌住院患者治疗前7.5ml外周血,保存于4℃冰箱中,24小时内由益善生物技术公司使用“免疫去除结合纳米过滤法”行CTCs检测。同时检测血清中Cyfra21-1、CEA、Fg水平,收集并分析CTCs数目与转移灶数目、临床T分期、转移淋巴结的关系。结果:纳入患者中CTCs检出率为86.4%。CTCs数目与Cyfra21-1水平有相关关系,相关系数为0.365（P=0.003）;CTCs数目与Fg水平相关,相关系数为0.330（P=0.007）;CEA与CTCs两变量无明显相关性。两组患者的肿瘤转移器官数目大于3个的比例有显著差异。结论:两组患者Cyfra21-1异常增高率有显著差异,且CTCs数目与Cyfra21-1表达水平正相关,可由Cyfra21-1初步估计CTCs水平。而外周血中Fg异常增高率无显著差异,但二者表达水平呈明显正相关,因此分析CTCs数目需要考虑Fg表达水平的影响。     

Therapy-related myelodysplastic syndrome and acute myeloid leukemia following initial treatment with chemotherapy plus radioimmunotherapy for indolent non-Hodgkin lymphoma

Patients with indolent non-Hodgkin lymphoma (I-NHL) often receive multiple courses of cytotoxic chemotherapy over several years. Radioimmunotherapy (RIT) has become an important part of treatment for relapsed patients and tositumomab/lodine I-131 tositumomab is a promising regimen currently being incorporated into first-line therapy. While treatment-related myelodysplasia (tMDS) and acute myeloid leukemia (tAML) are well-known, poor-prognosis complications of conventional chemotherapy and radiation therapy, they have not been previously observed as a consequence of initial treatment with RIT-based regimens. We describe four patients with tMDS/tAML who received a sequential chemotherapy and tositumomab/lodine I-131 tositumomab program as their initial and only lymphoma treatment. Our findings suggest that the potential risk of these important complications must be considered in the development of this novel therapeutic strategy in the first-line setting.     

Expression of MUC-1 epitopes on normal bone marrow: implications for the detection of micrometastatic tumor cells.

PURPOSE: The expression of the carcinoma-associated mucin MUC-1 is thought to be restricted to epithelial cells and is used for micrometastatic tumor cell detection in patients with solid tumors, including those with breast cancer. Little is known, however, about the expression of MUC-1 epitopes in normal hematopoietic cells. MATERIALS AND METHODS: MUC-1 expression was analyzed by flow cytometry and immunocytology on bone marrow (BM) mononuclear cells and purified CD34+ cells from healthy volunteers, using different anti-MUC-1-specific monoclonal antibodies. In addition, Western blotting of MUC-1 proteins was performed. RESULTS: Surprisingly, 2% to 10% of normal human BM mononuclear cells expressed MUC-1, as defined by the anti-MUC-1 antibodies BM-2 (2E11), BM-7, 12H12, MAM-6, and HMFG-1. In contrast, two antibodies recognizing the BM-8 and the HMFG-2 epitopes of MUC-1 were not detected. MUC-1+ cells from normal BM consisted primarily of erythroblasts and normoblasts. In agreement with this, normal CD34+ cells cultured in vitro to differentiate into the erythroid lineage showed a strong MUC-1 expression on day 7 proerythroblasts. Western blotting of these cells confirmed that the reactive species is the known high molecular weight MUC-1 protein. CONCLUSION: Our data demonstrate that some MUC-1 epitopes are expressed on normal BM cells and particularly on cells of the erythroid lineage. Hence the application of anti-MUC-1 antibodies for disseminated tumor cell detection in BM or peripheral blood progenitor cells may provide false-positive results, and only carefully evaluated anti-MUC-1 antibodies (eg, HMFG-2) might be selected. Furthermore, MUC-1-targeted immunotherapy in cancer patients might be hampered by the suppression of erythropoiesis.     

Tumor oxygenation after radiotherapy, chemotherapy, and/or hyperthermia predicts tumor free survival

PURPOSE: To investigate the influence of different treatment modalities (radiotherapy, chemotherapy, and hyperthermia) on the oxygenation of human tumor xenografts and to correlate it with the tumoricidal effect we conducted this study. METHODS AND MATERIALS: Human-derived head-and-neck squamous cell carcinoma xenografts (implanted in nude mice/nine groups of 10 mice) were treated with various treatment modalities and combinations of them (radiation with 5 x 2 or 10 x 2 Gy, hyperthermia at 41 degrees C or 41.8 degrees C, chemotherapy with ifosfamide [32 mg/kg] or cisplatin [2 mg/kg]). The tumor volume was evaluated 3 times per week until Day 60. Tumor pO(2) was measured at Day 1, 5, 8, and 12 with a polarographic pO(2) histograph. RESULTS: Within treatment time (maximum, 10 days) the median pO(2) increased in all groups (except the control group), concomitantly the fraction of measurements of pO(2) that were less than 10 mm Hg showed a constant decrease (p < or = 0.001). The highest difference between the median pO(2) values and the fraction of measurements of pO(2) that were less than 10 mm Hg at the start and 1 week after the end of therapy occurred in the groups with radiochemothermotherapy (triple-modality therapy; p< or = 0.001). At Day 60, the highest rate of complete remissions was observed in the triple-modality therapy groups. CONCLUSION: Tumor oxygenation under a single or combined cancer treatment is correlated with treatment efficacy in terms of complete remissions at Day 60. The posttherapeutic fraction of measurements of pO(2) that were less than 10 mm Hg correlates even better with the long term tumor free survival than the median pO(2) values or the pretherapeutic fraction of measurements of pO(2) that were less than 10 mm Hg.     

Combination therapy: lonidamine, hyperthermia, and chemotherapy against the RIF-1 tumor in vivo.

Lonidamine enhances the cytotoxicity in vitro of several conventional antitumor drugs as well as that of hyperthermia (HT). We have investigated the possibility that such enhancement can also be demonstrated in vivo against the RIF-1 tumor system. Two assays were used to examine antitumor activity: tumor growth delay and clonogenicity of cells obtained from tumors from treated animals. We used drug (and HT) doses that by themselves did not achieve significant cell killing. The drugs whose interaction with lonidamine was tested were: cis-diamminedichloroplatinum (CDDP), mitomycin C (MMC), bleomycin, 5-fluorouracil, and nitrosourea. Of these only CDDP and MMC yielded positive data. Both assays gave essentially the same results, showing that antitumor activity reflected direct cell killing. CDDP and MCC activity was also enhanced by HT. When we combined all three modalities, however, the results of the trimodality therapies were no better than that of individual bimodality treatments. These last results suggest that lonidamine and HT have similar mechanisms, most likely inhibition of repair of DNA damage. Our data do suggest that lonidamine may have a role in multidrug therapies that include either CDDP or MMC as a component of the treatments.     

U14荷瘤小鼠放化疗前后 IMP3蛋白的表达及意义

目的：探讨胰岛素样生长因子Ⅱ mRNA 结合蛋白3（insulin －like growth factor Ⅱ mRNA binding pro-tein 3,IMP3／IGF2BP3）在宫颈鳞癌实验动物模型（U14荷瘤小鼠）肿瘤组织放化疗前后中的表达变化。方法：将 U14细胞接种于昆明小鼠腹腔,形成腹水瘤模型,再抽取腹水,调整细胞数后,接种于昆明小鼠皮下,建立U14荷瘤小鼠实体瘤模型40例。根据成瘤后处理方式不同,将小鼠分为四组：对照组、化疗组、放疗组和放化疗组,测量不同组别小鼠肿瘤体积变化、抑瘤率,采用蛋白质印迹法（Western blot）检测不同处理组小鼠子宫颈鳞癌组织中 IMP3蛋白的表达水平。结果：与对照组相比,化疗组、放疗组、放化疗组肿瘤体积呈下降趋势,且放化疗后下降最明显（P ＜0．05）,抑瘤率逐渐递增（P ＜0．05）;IMP3蛋白在以上各组的平均相对灰度值分别为4．516±0．706、2．787±0．646、2．455±0．663、1．109±0．086,蛋白表达量逐渐下降,对照组明显高于治疗组,并且单纯放疗或化疗高于联合放化疗组,差异有明显的统计学意义（P ＜0．05）。结论：IMP3在小鼠子宫颈鳞癌组织中高表达;放化疗处理可降低其表达,下降量可在一定程度上反映治疗效果。     

CYFRA21-1、NSE和CEA在肺癌中的表达

目的分析CYFRA21－1、NSE和CEA在不同组织类型的肺癌中表达的差异。方法采用放免分析对13例正常人和37例肺癌患者进行血清CYFRA21-1、NSE和CEA的检测,统计分析各项指标在正常人、鳞癌、腺癌和小细胞肺癌患者之间的差别。结果CYFRA21-1在鳞癌、腺癌患者中表达水平较高,与小细胞肺癌及正常人有显著性差异;NSE在小细胞肺癌患者中表达水平较高,与鳞癌、腺癌及正常人有显著性差异;CEA在腺癌患者中表达水平较高,与鳞癌、小细胞肺癌及正常人有显著性差异。结论CYFRA21-1、NSE和CEA在肺癌中的表达与肺癌的组织类型密切相关。     

Deregulated expression of the human tumor marker CEA and CEA family member CEACAM6 disrupts tissue architecture and blocks colonocyte differentiation

Human carcinoembryonic antigen (CEA) and the CEA family member CEACAM6 (formerly nonspecific cross-reacting antigen [NCA]) function in vitro, at least, as homotypic intercellular adhesion molecules and, in model systems, can block the terminal differentiation and anoikis of several different cell types. We have recently demonstrated that the increased cell surface levels of CEA and CEACAM6 in purified human colonocytes from freshly excised, well to poorly differentiated colon carcinomas are inversely correlated with the degree of cellular differentiation. Thus, deregulated expression of CEA/CEACAM6 could directly contribute to colon tumorigenesis by the inhibition of terminal differentiation and anoikis. Evidence against this view includes the common observation of increased CEA/CEACAM6 expression as normal colonocytes differentiate in their migration up colonic crypt walls. We report here the direct effects of deregulated overexpression of CEA/CEACAM6, at levels observed in colorectal carcinomas, on the differentiation of two human colonic cell lines, SW-1222 and Caco-2. Stable transfectants of both of these cell lines that constitutively express 10- to 30-fold higher cell surface levels of CEA/CEACAM6 than endogenous levels failed to polarize and differentiate into glandular structures in monolayer or 3D culture or to form colonic crypts in a tissue architecture assay in nude mice. In addition, these transfectants were found to exhibit increased tumorigenicity in nude mice. These results thus support the contention that deregulated overexpression of CEA and CEACAM6 could provide a tumorigenic contribution to colon carcinogenesis.