Combined staining of TAG-72, MUC1, and CA125 improves labeling sensitivity in ovarian cancer: antigens for multi-targeted antibody-guided therapy.

Single antigen-targeted intraperitoneal radioimmunotherapy for ovarian cancer has shown limited success. Due to the heterogeneous expression of tumor antigens on cancer cells, a multi-antigen targeting approach appears logical to augment the therapeutic efficacy of antibody-guided therapy. In the interest of developing this novel approach, ovarian cancer tissue microarray slides containing cancer and benign/non-neoplastic tissue samples (n=92) were processed for single-, double-, and triple-antigen labeling using antibodies for the tumor-associated antigens TAG-72, MUC1, and CA125. Among all ovarian cancer types, 72%, 61%, and 50% of the samples showed immunolabeling for TAG-72, MUC1, and CA125, respectively. Expression level of these antigens was significantly (p<0.005) higher in advanced stage carcinomas compared with early stage. Of the 48 epithelial ovarian cancer samples, individual anti-TAG-72, MUC1, and CA125 antibody probing showed labeling in 89.5%, 87.5%, and 73.0% of the cases, respectively. In the majority of the cancer samples (>70%), a heterogeneous labeling pattern was observed (only 30-40% of the cancer cells within the sample were labeled). However, upon combining the three antigens (triple-antigen labeling), 98% of the epithelial ovarian cancer samples were labeled and >95% of the cancer cells within each sample were labeled. Our data indicate that the heterogeneous expression of cancer antigens appears to be a major obstacle in antibody-guided therapy, and this can be overcome by multiple antigen targeting. Therapeutic efficacy of antibody-guided therapy for ovarian cancer treatment will be enhanced by the combined targeting of TAG-72, MUC1, and CA125.     

Solution structure of the SEA domain from the murine homologue of ovarian cancer antigen CA125 (MUC16)

Human CA125, encoded by the MUC16 gene, is an ovarian cancer antigen widely used for a serum assay. Its extracellular region consists of tandem repeats of SEA domains. In this study we determined the three-dimensional structure of the SEA domain from the murine MUC16 homologue using multidimensional NMR spectroscopy. The domain forms a unique alpha/beta sandwich fold composed of two alpha helices and four antiparallel beta strands and has a characteristic turn named the TY-turn between alpha1 and alpha2. The internal mobility of the main chain is low throughout the domain. The residues that form the hydrophobic core and the TY-turn are fully conserved in all SEA domain sequences, indicating that the fold is common in the family. Interestingly, no other residues are conserved throughout the family. Thus, the sequence alignment of the SEA domain family was refined on the basis of the three-dimensional structure, which allowed us to classify the SEA domains into several subfamilies. The residues on the surface differ between these subfamilies, suggesting that each subfamily has a different function. In the MUC16 SEA domains, the conserved surface residues, Asn-10, Thr-12, Arg-63, Asp-75, Asp-112, Ser-115, and Phe-117, are clustered on the beta sheet surface, which may be functionally important. The putative epitope (residues 58-77) for anti-MUC16 antibodies is located around the beta2 and beta3 strands. On the other hand the tissue tumor marker MUC1 has a SEA domain belonging to another subfamily, and its GSVVV motif for proteolytic cleavage is located in the short loop connecting beta2 and beta3.     

NBR1 interacts with fasciculation and elongation protein zeta-1 (FEZ1) and calcium and integrin binding protein (CIB) and shows developmentally restricted expression in the neural tube.

NBR1 (named as next to BRCA1) was originally cloned as a candidate gene for the ovarian cancer antigen CA125, using expression cloning with the anti-CA125 Ig, OC125. NBR1 has been of interest due to its position close to BRCA1, although no involvement in breast or ovarian cancer has been demonstrated. Recently, the antigen CA125 has been cloned, and identified as a new mucin, MUC16, entirely different from NBR1. The function of NBR1 remains unknown. To investigate its function, a yeast two-hybrid study was performed to identify interacting protein partners that may reflect a biological role for this protein. Here, we show that NBR1 interacts with two proteins; fasciculation and elongation protein zeta-1 (FEZ1), a PKCzeta interacting protein, and calcium and integrin binding protein (CIB), which is associated with polo-like kinases Fnk/Snk and the Alzheimer's disease presenilin 2 protein. Co-transfection of FEZ1 and NBR1 showed overlapping localization in the cytoplasm, whereas coexpression of NBR1 and CIB resulted in a shift of CIB protein expression from the nucleus to the perinuclear compartment. FEZ1 is highly expressed in the brain and in situ hybridization analysis of Nbr1 showed that its expression is also regulated in the murine brain during development. These data suggest that NBR1 may function, through interaction with CIB and FEZ1 in cell signalling pathways, with a developmentally restricted expression suggesting a possible role in neural development.     

CA125/MUC16 interacts with Src family kinases,and over-expression of its C-terminal fragment in human epithelial cancer cells reduces cell–cell adhesion

MUC16/CA125 is over-expressed in human epithelial tumors including ovarian, breast and some other carcinomas. The purpose of this study is to investigate how cell surface MUC16 is functionally involved in tumor progression, with a special focus on the role of its cytoplasmic tail. Forced expression of C-terminal MUC16 fragment (MUC16C) in epithelial cancer cells increased cell migration. We found that MUC16C directly interacted with Src family kinases (SFKs). Notably, localizations of E-cadherin and β-catenin at the cell–cell contacts were more diffuse in MUC16C transfectants compared with mock transfectants. Furthermore, MUC16C transfectants showed reduced Ca²⁺-dependent cell–cell adhesion, but the treatment of cells with PP2, a SFKs inhibitor, restored this. Because cell surface MUC16 is also associated with the E-cadherin/β-catenin complex, the over-expression of MUC16 and its interaction with SFKs may enhance SFKs-induced deregulation of E-cadherin. Thus, our results suggest a role for cell surface MUC16 in cell–cell adhesion of epithelial cancer cells.     

Soluble MUC1 and serum MUC1-specific antibodies are potential prognostic biomarkers for platinum-resistant ovarian cancer

MUC1 (CA15-3) and MUC16 (CA125) tumor-associated antigens are upregulated in ovarian cancer and can be detected in patients’ sera by standardized tests. We postulated that increased MUC1 and MUC16 antigens augment antibody responses in platinum-resistant ovarian cancer patients and that the frequency and intensity of these responses can be used as immune biomarkers of treatment response and disease outcome. We measured MUC1 and MUC16 tumor expression by immunohistochemistry (IHC), assessed serum antigenic levels and quantitated circulating antibodies by ELISA in a cohort of 28 ovarian cancer patients with platinum-resistant or platinum-refractory ovarian cancer, and treated with intraperitoneal (IP) interleukin-2 (IL-2). MUC1 and MUC16 were overexpressed in tumor samples and showed differential distribution profiles. Serum MUC1 (CA15-3) measurements were elevated in all patients and significantly correlated with increased risk of death (P = 0.003). MUC1-specific IgM and IgG anitbodies were found in 92 and 50% of cases, respectively. Patients with progressive disease had higher mean anti-MUC1 IgG than responders at both early (P = 0.025) and late (P = 0.022) time points during IP IL-2 treatment. Anti-MUC1 IgM antibodies inversely correlated with overall survival at both early (P = 0.052) and late (P = 0.009) time points. In contrast to MUC1, neither soluble MUC16 nor MUC16-specific antibodies were significantly associated with clinical response or overall survival in this study. Increased serum MUC1 and high anti-MUC1 antibody levels are prognostic for poor clinical response and reduced overall survival in platinum-resistant or platinum-refractory ovarian cancer.     

Free CA125 promotes ovarian cancer cell migration and tumor metastasis by binding Mesothelin to reduce DKK1 expression and activate the SGK3/FOXO3 pathway

Objective: CA125/MUC16 is an O-glycosylated protein that is expressed on the surfaces of ovarian epithelial cells. This molecule is a widely used tumor-associated marker for diagnosis of ovarian cancer. Recently, CA125 was shown to be involved in ovarian cancer metastasis. The purpose of this study was to investigate the mechanism of CA125 during ovarian cancer metastasis.Methods: We analyzed the Oncomine and CSIOVDB databases to determine the expression levels of DKK1 in ovarian cancer. DKK1 expression levels were upregulated or downregulated and applied with CA125 to Transwell and Western blot assays to ascertain the underlying mechanism by which CA125 stimulates cell migration via the SGK3/FOXO3 pathway. Anti-mesothelin antibodies (anti-MSLN) were used to block CA125 stimulation. Then the expression levels of DKK1were tested by enzyme-linked immunosorbent assay (ELISA) to eliminate the blocking effect of anti-MSLN to CA125 stimulation. Xenograft mouse models were used to detect the effects of CA125 and anti-MSLN on ovarian cancer metastasis in vivo.Results: DKK1 levels were downregulated in ovarian tumor tissues according to the analyses of two databases and significantly correlated with FIGO stage, grade and disease-free survival in ovarian cancer patients. DKK1 levels were downregulated by CA125 stimulation in vitro. Overexpression of DKK1 reversed the ability of exogenous CA125 to mediate cell migration by activating the SGK3/FOXO3 signaling pathway. Anti-MSLN abrogated the DKK1 reduction and increased the apoptosis of ovarian cancer cells. The use of anti-MSLN in xenograft mouse models significantly reduced tumor growth and metastasis accelerated by CA125.Conclusions: These experiments revealed that the SGK3/FOXO3 pathway was activated, wherein decreased expression of DKK1 was caused by CA125, which fuels ovarian cancer cell migration. Mesothelin is a potential therapeutic target for the treatment of ovarian cancer metastasis.     

Pathobiological implications of MUC16 expression in pancreatic cancer

MUC16 (CA125) belongs to a family of high-molecular weight O-glycosylated proteins known as mucins. While MUC16 is well known as a biomarker in ovarian cancer, its expression pattern in pancreatic cancer (PC), the fourth leading cause of cancer related deaths in the United States, remains unknown. The aim of our study was to analyze the expression of MUC16 during the initiation, progression and metastasis of PC for possible implication in PC diagnosis, prognosis and therapy. In this study, a microarray containing tissues from healthy and PC patients was used to investigate the differential protein expression of MUC16 in PC. MUC16 mRNA levels were also measured by RT-PCR in the normal human pancreatic, pancreatitis, and PC tissues. To investigate its expression pattern during PC metastasis, tissue samples from the primary pancreatic tumor and metastases (from the same patient) in the lymph nodes, liver, lung and omentum from Stage IV PC patients were analyzed. To determine its association in the initiation of PC, tissues from PC patients containing pre-neoplastic lesions of varying grades were stained for MUC16. Finally, MUC16 expression was analyzed in 18 human PC cell lines. MUC16 is not expressed in the normal pancreatic ducts and is strongly upregulated in PC and detected in pancreatitis tissue. It is first detected in the high-grade pre-neoplastic lesions preceding invasive adenocarcinoma, suggesting that its upregulation is a late event during the initiation of this disease. MUC16 expression appears to be stronger in metastatic lesions when compared to the primary tumor, suggesting a role in PC metastasis. We have also identified PC cell lines that express MUC16, which can be used in future studies to elucidate its functional role in PC. Altogether, our results reveal that MUC16 expression is significantly increased in PC and could play a potential role in the progression of this disease.     

Binding of ovarian cancer antigen CA125/MUC16 to mesothelin mediates cell adhesion

Mesothelin is a glycosylphosphatidylinositol-linked cell surface molecule expressed in the mesothelial lining of the body cavities and in many tumor cells. Based on the finding that a soluble form of mesothelin specifically binds to ovarian carcinoma cell line OVCAR-3, we isolated cDNAs encoding a mesothelin-binding protein by expression cloning. The polypeptides encoded by the two cloned cDNA fragments matched to portions of CA125, an ovarian cancer antigen and a giant mucin-like glycoprotein present at the surface of tumor cells. By flow cytometric analysis and immunoprecipitation, we demonstrate that CA125 binds to mesothelin in a specific manner. Binding of CA125 to membrane-bound mesothelin mediates heterotypic cell adhesion as anti-mesothelin antibody blocks binding of OVCAR-3 cells expressing CA125 to an endothelial-like cell line expressing mesothelin. Finally, we show that CA125 and mesothelin are co-expressed in advanced grade ovarian adenocarcinoma. Taken together, our data indicate that mesothelin is a novel CA125-binding protein and that CA125 might contribute to the metastasis of ovarian cancer to the peritoneum by initiating cell attachment to the mesothelial epithelium via binding to mesothelin.     

A Binding Domain on Mesothelin for CA125/MUC16

Ovarian cancer and malignant mesothelioma frequently express both mesothelin and CA125 (also known as MUC16) at high levels on the cell surface. The interaction between mesothelin and CA125 may facilitate the implantation and peritoneal spread of tumors by cell adhesion, whereas the detailed nature of this interaction is still unknown. Here, we used truncated mutagenesis and alanine replacement techniques to identify a binding site on mesothelin for CA125. We examined the molecular interaction by Western blot overlay assays and further quantitatively analyzed by enzyme-linked immunosorbent assay. We also evaluated the binding on cancer cells by flow cytometry. We identified the region (296–359) consisting of 64 amino acids at the N-terminal of cell surface mesothelin as the minimum fragment for complete binding activity to CA125. We found that substitution of tyrosine 318 with an alanine abolished CA125 binding. Replacement of tryptophan 321 and glutamic acid 324 with alanine could partially decrease binding to CA125, whereas mutation of histidine 354 had no effect. These results indicate that a conformation-sensitive structure of the region (296–359) is required and sufficient for the binding of mesothelin to CA125. In addition, we have shown that a single chain monoclonal antibody (SS1) recognizes this CA125-binding domain and blocks the mesothelin-CA125 interaction on cancer cells. The identified CA125-binding domain significantly inhibits cancer cell adhesion and merits evaluation as a new therapeutic agent for preventing or treating peritoneal malignant tumors.Ovarian cancer largely is confined to the peritoneal cavity for much of its natural history (1). Peritoneal mesothelioma is a highly invasive tumor originating from the mesothelial linings of the peritoneum (2). The development of effective drug regimens against ovarian cancer and mesothelioma has proven extremely difficult.Mesothelin was first identified in 1992 by the monoclonal antibody (mAb)² K1 that was generated by the immunization of mice with human ovarian carcinoma (OVCAR-3) cells (3). The mesothelin gene encodes a 71-kDa precursor protein that is processed to a 40-kDa protein termed mesothelin, which is a glycosylphosphatidylinositol (GPI)-anchored glycoprotein present on the cell surface (4). Mesothelin is a differentiation antigen that is present on a restricted set of normal adult tissues such as the mesothelium. In contrast, it is overexpressed in a variety of cancers including mesothelioma, ovarian cancer, and pancreatic cancer (5). In addition, mesothelin is also expressed on the surface of non-small cell lung cancer cells (6, 7), especially most lung adenocarcinomas (8).We and others have shown that mesothelin is shed from tumor cells (9, 10), and antibodies specific for mesothelin are elevated in the sera of patients with mesothelioma and ovarian cancer (11). Shed serum mesothelin has been approved by the United States Food and Drug Administration (FDA) as a new diagnostic biomarker in mesothelioma. In a Phase I clinical study of an intrapleural interferon-β gene transfer using an adenoviral vector in patients with mesotheliomas, we found that antitumor immune responses targeting mesothelin were elicited in several patients (12). A recent study indicated that anti-mesothelin antibodies and circulating mesothelin relate to the clinical state in ovarian cancer patients (13). Pastan and colleagues (14) developed an immunotoxin (SS1P) with a Fv for mesothelin. Two Phase I clinical trials were completed at the National Cancer Institute (National Institutes of Health, Bethesda, MD) and there was sufficient antitumor activity of SS1P to justify a Phase II trial. A chimeric antibody containing the mouse SS1 Fv for mesothelin was also developed and is currently examined in a Phase I clinical trial for ovarian cancer, mesothelioma, pancreatic cancer, and non-small cell lung cancer (15).Mucins are heavily glycosylated proteins found in the mucus layer or at the cell surface of many epitheliums (16). There are two structurally distinct families of mucins, secreted and membrane-bound forms. CA125 (also known as MUC16) was first identified in 1981 by OC125, a mAb that had been developed from mice immunized with human ovarian cancer cells (17). The first cDNA clones were reported in 2001 (18, 19). CA125 is a very large membrane-bound cell surface mucin, with an average molecular mass between 2.5 and 5 million daltons. It is also heavily glycosylated with both O-linked and N-linked oligosaccharides (20). The peptide backbone of CA125 is composed of the N-terminal region, extensive Ser/Thr/Pro-rich tandem repeats (TR) with 156 amino acids each with both N- and O-glycosylations, a SEA domain with high levels of O-glycosylation and a C-terminal region with a short cytoplasmic tail (19). The SEA domain was first identified as a module commonly found in sea urchin sperm protein, enterokinase and agrin (21, 22). The significance of the SEA domain in CA125 is not clear.CA125 was originally used as a biomarker in ovarian cancer due to its high expression in ovarian carcinomas and that it is shed into the serum (23). A majority (88%) of mesotheliomas are also CA125 positive on the cell membrane (24). It was shown that 25% of peritoneal mesotheliomas have high CA125 expression (25). The intensity of CA125 membranous expression is indistinguishable between ovarian carcinomas and peritoneal mesotheliomas. Gene expression analysis using the SAGE tag data base has shown that mesothelioma has the second highest co-expression of CA125 and mesothelin after ovarian cancer (26). Rump and colleagues (26) have shown that mesothelin binds to CA125 and that this interaction may mediate cell adhesion. Scholler et al. (27) recently showed that CA125/mesothelin-dependent cell attachment could be blocked with anti-CA125 antibodies. Because mesothelin is present on peritoneal mesothelium, there may be an important role for the mesothelin-CA125 interaction in the tumorigenesis of ovarian cancer and mesothelioma in the peritoneal cavity. The mesothelin binding site on CA125 may lie within the 156-amino acid TR units, indicating multimeric binding of mesothelin to CA125. It has been found that the extraordinarily abundant N-glycans on CA125, presumably in the TR region, are required for binding to both glycosylated and non-glycosylated mesothelin (28).Here, we identified the binding site of CA125 on mesothelin by use of truncated mutagenesis and alanine replacement approaches. We measured binding qualitatively by Western blot overlay assays and quantitatively by enzyme-linked immunosorbent assay (ELISA). We also evaluated the interaction of CA125 and mesothelin on cancer cells by flow cytometry. Furthermore, we have shown that a single chain mAb (SS1) recognized the CA125-binding domain and blocked the mesothelin-CA125 interaction on cancer cells. The identified CA125-binding domain-Fc fusion protein also significantly inhibited cancer cell adhesion. Our results suggest that conformation-sensitive structures of the region (296–359) are required and sufficient for specific binding of mesothelin to CA125. The domain proteins or the antibodies that block the mesothelin-CA125 interaction merit evaluation as new therapeutic agents in treating peritoneal malignant tumors.     

Enhanced discrimination of malignant from benign pancreatic disease by measuring the CA 19-9 antigen on specific protein carriers

The CA 19-9 assay detects a carbohydrate antigen on multiple protein carriers, some of which may be preferential carriers of the antigen in cancer. We tested the hypothesis that the measurement of the CA 19-9 antigen on individual proteins could improve performance over the standard CA 19-9 assay. We used antibody arrays to measure the levels of the CA 19-9 antigen on multiple proteins in serum or plasma samples from patients with pancreatic adenocarcinoma or pancreatitis. Sample sets from three different institutions were examined, comprising 531 individual samples. The measurement of the CA 19-9 antigen on any individual protein did not improve upon the performance of the standard CA 19-9 assay (82% sensitivity at 75% specificity for early-stage cancer), owing to diversity among patients in their CA 19-9 protein carriers. However, a subset of cancer patients with no elevation in the standard CA 19-9 assay showed elevations of the CA 19-9 antigen specifically on the proteins MUC5AC or MUC16 in all sample sets. By combining measurements of the standard CA 19-9 assay with detection of CA 19-9 on MUC5AC and MUC16, the sensitivity of cancer detection was improved relative to CA 19-9 alone in each sample set, achieving 67-80% sensitivity at 98% specificity. This finding demonstrates the value of measuring glycans on specific proteins for improving biomarker performance. Diagnostic tests with improved sensitivity for detecting pancreatic cancer could have important applications for improving the treatment and management of patients suffering from this disease.     

Roles of CA125 in diagnosis,prediction, and oncogenesis of ovarian cancer

After it was discovered approximately 40 years ago, carbohydrate antigen 125 (CA125) became the most widely used and concerning biomarker in ovarian cancer screening. However, there is still controversy about its role in clinical practice. CA125 is not sufficiently reliable in diagnosis to screen for early-stage ovarian cancer. On the other hand, CA125 has been a valuable indicator for evaluating chemotherapeutic efficacy and prognosis. We still do not know much about its biological role, and several studies have indicated that this marker participates in the occurrence and development of ovarian cancer. Currently, an increasing number of scholars have begun to pay attention to CA125-targeted treatment strategies. In the interest of better design and development of anticancer therapies, a renewed and systematic understanding of the roles of CA125 in diagnosis, prediction, and tumorigenesis is warranted.     

Study of lung cancer regulatory network that involves erbB4 and tumor marker gene

Our purpose is to screen out serum tumor markers closely correlated to the nature of solitary pulmonary nodule (SPN) and to draw a regulatory network containing genes correlated to lung cancer. Two hundred and sixty cases of SPN patients confirmed through pathological diagnosis were collected as subjects, factors closely correlated to the nature of SPN were screened out from eight tumor markers through Fisher discriminant method, and functional annotation and pathway analysis were conducted on erbB4 as well as its tumor marker genes by GO and KEGG databases. Four key tumor markers: CYFRA21-1, CA125, SCC-Ag and CA153 were successfully screened out and the first three proteins’ corresponding gene were KRT19, MUC16 and SERPINB3 while that of CA153 was not found. GO analysis on erbB4, KRT19, MUC16 and SERPINB3 showed that they covered three domains, cell components, molecular function and biological process; meanwhile, combined with KEGG database and based on signal pathway of erbB4, a regulatory network of lung cancer cells escaping from apoptosis was successfully made. This study indicates that serum tumor marker genes play an important role in the occurrence and development of lung cancer, besides, this study primarily discussed the molecular mechanism of these tumor markers in predicting tumor, which provides a basis for in-depth information about lung cancer.     

Isolation of the MurineNbr1Gene Adjacent to the MurineBrca1Gene

The humanNBR1cDNA has previously been identified using polyclonal sera to CA125, an ovarian tumor antigen used in monitoring ovarian cancer. The gene was mapped to theBRCA1region on chromosome 17q21 and subsequently found to lie in close proximity to the recently identifiedBRCA1gene. The NBR1 protein has a B-box motif but the function of the protein is as yet unknown. To investigate the function and importance of this gene, we have studied the conservation of this gene in other species and in particular in the mouse. We have isolated murineNbr1cDNA and genomic clones. Translation of the cDNA sequence indicates that the protein is highly conserved, being 89% similar and 84% identical to the human. Analysis of the murineNbr1genomic clones indicates that it maps less than 1 kb from theBrca1gene and that, unlike that in human, this region is not duplicated.     

卵巢癌患者血清HE4、CA125、CA72-4及炎性因子水平检测的临床价值评估

目的:探讨血清人附睾蛋白4(HE4)、糖类抗原125(CA125)、糖类抗原72-4(CA72-4)及炎性因子IL-6、IL-8、IL-17水平检测对卵巢癌患者的临床价值。方法:选取2015年2月至2017年2月我院收治的卵巢肿瘤患者81例,包括卵巢恶性肿瘤组39例及卵巢良性肿瘤组42例,另选42例健康人作正常对照组,比较各组及临床不同分期卵巢癌患者血清HE4、CA125、CA72-4、IL-6、IL-8、IL-17水平的差异。结果:卵巢恶性肿瘤组血清HE4、CA125、CA72-4、IL-6、IL-8及IL-17水平均显著高于对照组(P0.05),而卵巢良性肿瘤组与对照组以上指标比较差异并无统计学意义(P0.05);随卵巢癌分期增加,患者血清HE4、CA125、CA72-4、IL-6、IL-8及IL-17水平呈上升趋势,以血清HE4、CA125及IL-17增加较为显著(P0.05),而Ⅰ期、Ⅱ期及Ⅲ期卵巢癌患者血清CA72-4、IL-6及IL-8在增加并不明显(P0.05)。结论:卵巢癌患者血清HE4、CA125、CA72-4、IL-6、IL-8及IL-17水平较高,且随临床分期增加呈上升趋势,以上指标可能有助于卵巢癌的诊断、治疗及病情评估。     

上皮性卵巢癌患者CT、MRI影像学特征及与血清标志物CEA、CA199、CA125水平的相关性研究

摘要 目的：探讨上皮性卵巢癌患者电子计算机断层扫描（CT）、磁共振成像（MRI）影像学特征及与血清标志物癌胚抗原（CEA）、糖类抗原199（CA199）、糖类抗原125（CA125）水平的相关性。方法：回顾性分析2014年4月-2020年2月于我院83例诊断为上皮性卵巢癌患者的CT、MRI影像学资料,以手术病理结果作为金标准。分析患者的CT、MRI影像学特征,检测患者血清CEA、CA199、CA125水平,评价患者CT、MRI影像学特征与血清CEA、CA199、CA125水平的相关性。结果：上皮性卵巢癌肿瘤横截面最大径为14.2mm-121.7mm,平均（18.6±4.3）mm,上皮性卵巢癌以混杂密度/信号为主,形态不规则,病灶多为囊实性,可见壁结节及分隔改变,增强后可见分隔或壁结节明显强化,可伴有腹水、腹膜转移、淋巴结转移。血清CEA、CA199、CA125水平分别为（66.35±7.52）ng/mL、（183.59±22.62）U/mL、（225.27±25.34）U/mL。上皮性卵巢癌边界清晰、不清晰的血清CA199、CA125水平组间差异有统计学意义（P＜0.05）;上皮性卵巢癌形态圆形/类圆形/椭圆形、分叶状、形态不规则的血清CA199、CA125水平组间差异有统计学意义（P＜0.05）;上皮性卵巢癌患者有壁结节、腹膜转移、淋巴结转移的血清CEA、CA199、CA125水平组间差异有统计学意义（P＜0.05）;其余CT、MRI影像学表现特征组间血清CEA、CA199、CA125水平差异无统计学意义（P＞0.05）。上皮性卵巢癌边界与血清CA125水平呈正相关（P＜0.05）,上皮性卵巢癌形态与血清CA199、CA125水平呈正相关（P＜0.05）,壁结节与血清CA125水平呈正相关（P＜0.05）,腹膜转移、淋巴结转移与血清CEA、CA199、CA125水平呈正相关（P＜0.05）,其余指标之间无明显相关性（P＞0.05）。结论：上皮性卵巢癌CT、MRI影像表现具有特征性,血清CEA、CA199、CA125水平的检测有助于对早期上皮性卵巢癌的诊断以及不同病理类型的判断,CT、MRI影像学特征与血清CEA、CA199、CA125水平具有相关性,可判断疾病的进展及患者预后情况,对指导临床综合治疗及评估患者预后可提供客观依据。     

The nucleotide sequence and derived amino acid sequence of cDNA coding for mouse carbonic anhydrase II

The nucleotide sequence of a clone containing mouse carbonic anhydrase (CA) cDNA in pBR322 has been determined. The cloned cDNA contains all of the coding region except for nucleotides specifying the first eight amino acids, and all of the 3' noncoding region, which consists of 700 nucleotides. A cDNA clone was identified which contains an additional 54 bp at the 5' end, so that the complete amino acid sequence of mouse CA could be deduced. This sequence showed a 73-81% homology with other mammalian CA form II isozymes, 56-63% with form I isozymes, and 52-56% with form III isozymes. By examination of the amino acids which are unique and invariant for each isozyme, the mouse amino acid sequence was found to contain 16 of the 23 residues that are unique and invariant to mammalian CA form II isozymes, but only one or no residue for forms I and III, respectively.     

HE4和HE4/CA125并联诊断卵巢癌的系统评价

目的:通过Meta分析方法系统评价血清人附睾蛋白4(HE4)及HE4并联糖类抗原125(CA125)诊断卵巢癌的价值。方法:中国知网期刊数据库(CNKI)、中文科技期刊数据库(中国万方数据库)、中文电子期刊数据库及维普网等数据库检索2005年1月至2015年3月的相关文献,使用Meta-Disc软件进行Meta分析,分析异质性及处理后选择适当效应模型合并效应量,得出合并敏感性、合并特异性及合并似然比,制作SROC曲线并计算曲线下面积(AUC)。对HE4及HE4/CA125并联检测诊断卵巢癌的AUC应用Z检验进行比较。采用STATA13.0软件应用Egger检验法进行发表偏倚检测。结果:16篇文献符合纳入标准,将对照组分为健康人群和良性疾病组,经Meta分析后得出:以健康人群为对照,HE4和HE4/CA125并联检测卵巢癌的AUC分别为0.8611±0.0399、0.8959±0.0237,差异无统计学意义(Z值=0.749871,P0.05)。以良性疾病组为对照,HE4和HE4/CA125并联检测卵巢癌的AUC分别为0.9443±0.0153、0.9328±0.0132,差异无统计学意义(Z值=0.569105,P0.05)。结论:HE4/CA125并联检测提高了卵巢癌检测的灵敏度,而单独检测HE4有良好的特异度,两者对卵巢癌诊断均有较高的AUC,但HE4与HE4/CA125并联检测的诊断价值差异并无统计学意义。     

Human MUC4 mucin cDNA and its variants in pancreatic carcinoma

The human MUC4 gene is not expressed in normal pancreas; however, its dysregulation results in high levels of expression in pancreatic tumors. To investigate the tumor-associated expression, MUC4 cDNA was cloned from a human pancreatic tumor cell line cDNA expression library using a polyclonal antibody raised against human deglycosylated mucin and RT-PCR. Pancreatic MUC4 cDNA shows differences in 12 amino acid residues in the non-tandem repeat coding region with no structural rearrangement as compared with tracheal MUC4. The full-length MUC4 cDNA includes a leader sequence, a serine and threonine rich non-tandem repeat region, a central large tandem repeat domain containing 48 bp repetitive units, regions rich in potential N-glycosylation sites, two cysteine-rich domains, EGF-like domains, and a transmembrane domain. We also report the presence of a new EGF-like domain in MUC4 cDNA, located in the cysteine-rich region upstream from the first EGF-like domain. Four distinct splice events were identified in the region downstream of the central tandem repeat domain that generate three new MUC4 cDNA sequences (sv4, sv9, and sv10). The deduced amino acid sequences of two of these variants lack the transmembrane domain. Furthermore, two unique forms of MUC4 (MUC4/Y and MUC4/X) generated as a result of alternative splicing lack the salient feature of mucins, the tandem repeat domain. A high degree of polymorphism in the central tandem repeat region of MUC4 was observed in various pancreatic adenocarcinoma cell lines, with allele sizes ranging from 23.5 to 10.0 kb. MUC4 mRNA expression was higher in differentiated cell lines, with no detectable expression in poorly differentiated pancreatic tumor cell lines.