Cancer Antigen CA125 Research Articles

Características operativas del método de electroquimioluminiscencia para la medida de la proteína epididimaria humana (HE4) en el analizador Cobas e411 (Roche Diagnostics®)

IntroductionHuman epididymis protein 4 (HE4) is an emerging tumor marker in diagnosis of ovarian cancer. Together with the cancer antigen CA 125 it can improve sensitivity and specificity by calculating the risk of ovarian malignancy algorithm (ROMA) score. Our purpose is to validate performance characteristics of the electrochemiluminiscent Cobas e411 assay for HE4, including the endogenous interference study. Material and methodsTwo different levels of commercial control were analyzed for imprecision and trueness study during 20 days. Limit of blank and limit of detection was assessed with 20 replicates of a sample lacking HE4. Linearity was assessed by regression analysis mixing specimens, with high HE4 concentration and a devoid analyte one. In addition carry-over assay and endogenous interference study were conducted. ResultsThe imprecision obtained for each level were 1.7 and 2.8% for within run, 2.59 and 1.28%, for between run and 4.67 and 4.47% for total imprecision. The relative biases were 6.11 and 1.54% for each level. The limit of blank was 1.25 pmol/L and the limit of detection 2.21 pmol/L. The linearity study yielded the following regression equation y=-18.22+0.98x, demonstrating a linear behavior. There was not significant carry-over. The method was not affected by hemolysis, bilirrubin, neither lipemia. ConclusionsElectrochemiluminiscence method for HE4 showed an excellent imprecision and trueness. The limit of detection resulted lower than the declared by the manufacturer. Linearity was verified within the limits studied. In addition the assay is devoid of carry-over and free of significant endogenous interferences.

Core–shell gold–silver nanoparticles based impedimetric immunosensor for cancer antigen CA125

We report a simple and fast assay for cancer antigen CA125 quantification using gold–silver core–shell nanoparticles for ovarian cancer diagnosis and prognosis. The impedimetric analysis of CA125 using label-free and reagentless immunosensor could be used done in 20min. Furthermore, the directed immobilization of antibody onto the core–shell nanoparticles enabled linear response upto 1–150IU/mL (r2=0.994) as opposed to the maximum linear response of 1.0–100.0IU/mL reported for impedimetric immunosensors till date. The fabricated immunosensor displayed tolerable interference of 2.0–5% from serum components and retained 90% stability up to 20 days. Moreover, sensitivity of immunosensor fabricated using Au–Ag nanoparticles (190ΩIU−1mLcm−2) was almost three times that of gold nanoparticles based immunosensor (59ΩU−1mLcm−2).

Relationship of tumor marker CA125 and ovarian tumor stem cells: preliminary identification.

PurposeThe purpose of this study is to identify a prospective association between CA125 and tumorigenic ovarian cancer cells, using the new method of orthotopic transplantation (1).MethodAfter making the surgical ovarian cancer specimen into cell suspension, we separated the tumorigenic cells from the nontumorigenic cancer cells based on cell surface marker (cancer antigen CA125 and lineage markers) expression. We developed a SCID mice model in which the CA125+/ lineage- and CA125-/ lineage- cells were injected into ovarian parenchyma by use of a microinjector. As a measure of effectiveness of tumor-forming, tumor weight, abdominal distension, ascites volume and activity, subcutaneous fat were determined or observed. Immunohistochemistry was done to determine tumor cell markers.ResultsWe found that the cells of CA125+/ lineage- were able to form new tumors; whereas, an equal quantity of CA125-/lineage- cells failed to form any tumors. The new generated tumor contained additional CA125-/lineage- tumorigenic cells as well as the phenotypically diverse population of nontumorigenic cells. Quantities were judged to be significantly different P < 0.0001.ConclusionCA125+/ lineage- cells, which may be ovarian cancer stem cells, were the source for tumor recurrence. The strategies designed to target this cell population may lead to more effective therapies.

The Significance of CA15-3 in Breast Cancer Patients and its Relationship to HER-2 Receptor Status

Breast cancer is estimated to be the most common malignancy affecting women in Iraq. The cancer antigen CA 15-3 has been used as a possible serum marker of occult and recurrent breast carcinoma, either alone or in combination with other tumor markers such as HER2/neu, that has evolved as a major classifier of invasive breast cancer and target of therapy for the disease. ELISA, used to evaluate serum levels of CA15-3 and immuno-histochemistry staining technique, was used to establish the HER2/neu status. The results of this study indicate an increased level of CA15-3 in breast cancer patients (29.02+/-1.79 IU/ml) as compared to both women with benign tumor and healthy controls (13.78+/-1.24 and 8.92+/-0.48 IU/ml, respectively), and that this increase is associated to advanced stages. Patients with HER2/neu positive malignancies show elevated serum CA15-3 (37.09+/-2.55 IU/ml), as well as patients who developed recurrence (40.75+/-2.11 IU/ml). Our data study suggests that higher levels of CA 15-3 would be a reliable prognostic marker as they were directly related to advanced stages and recurrence. In addition persistent elevation of CA 15-3 was associated to HER2/neu positivity in breast cancer patients.

CA-125–indicated asymptomatic relapse confers survival benefit to ovarian cancer patients who underwent secondary cytoreduction surgery

BackgroundThere is no consensus regarding the management of ovarian cancer patients, who have shown complete clinical response (CCR) to primary therapy and have rising cancer antigen CA-125 levels but have no symptoms of recurrent disease. The present study aims to determine whether follow-up CA-125 levels can be used to identify the need for imaging studies and secondary cytoreductive surgery (CRS).MethodsWe identified 410 ovarian cancer patients treated at The University of Texas MD Anderson Cancer Center between 1984 and 2011. These patients had shown CCR to primary therapy. Follow-up was conducted based on the surveillance protocol of the MD Anderson Cancer Center. We used the Cox proportional hazards model and log-rank test to assess the associations between the follow-up CA-125 levels and secondary CRS and survival duration.ResultsThe CA-125 level of 1.68 × nadir was defined as the indicator of recurrent disease (p < 0.001). The specificity and sensitivity of this criterion were 82.9% and 85.6%, respectively, and the median lead-time of the CA-125 biochemical progression prior to clinically-defined relapse was 31 days (ranging from 1 to 391 days). The median number of the negative imaging studies for the clinical relapse findings in patients with a CA-125 level of < 1.68 × nadir was 3 (ranging from 0 to 24 times). The increase of CA-125 level at relapse was an independent predictor of overall and progression free survival in patients who had shown CCR to primary therapy (p = 0.04 and 0.02 respectively). The overall and progression free survival durations in patients with a CA-125 level ≤ 1.68 × nadir at relapse (69.4 and 13.8 months) were longer than those with a CA-125 level > 1.68 × nadir at relapse (55.7 and 10.4 months; p = 0.04 and 0.01, respectively). The overall and progression free survival duration of patients with asymptomatic relapse and underwent a secondary CRS was longer than that of patients with symptomatic relapse (p = 0.02 and 0.04 respectively).ConclusionsThe increase of serum CA-125 levels is an early warning of clinical relapse in ovarian cancer. Using CA-125 levels in guiding the treatment of patients with asymptomatic recurrent ovarian cancer, who have shown CCR to primary therapy, can facilitate optimal secondary CRS and extend the survival duration of the patients.

Recognition of Mesothelin by the Therapeutic Antibody MORAB-009: Structural and Mechanistic Insights

Mesothelin is a cell surface protein that is normally found in mesothelial cells lining the pleura, pericardium and peritoneum, but is aberrantly expressed at high level in a variety of cancers including mesothelioma, ovarian, pancreatic and lung cancers. Although the physiological function of mesothelin is unclear, studies have shown that it is capable of binding to the tumor antigen CA-125 (also known as MUC16), leading to cell adhesion and tumor metastasis. Since mesothelin is specifically expressed at a significantly higher level in malignant tumors, development of an antibody against mesothelin is therefore of major importance in the field of cancer therapy. MORAb-009 is a humanized monoclonal antibody against mesothelin currently under clinical trials. Animal experiments have shown that application of MORAb-009 in combination with chemotherapy leads to a marked reduction in tumor growth of mesothelin-expressing tumors. We show here that MORAb-009 recognizes a non-linear epitope that is contained in the first 64-residue fragment of the mesothelin. We further demonstrate that the recognition is sensitive to the loss of a disulfide bond linking residues Cys7 and Cys31. The crystal structure of the complex between the mesothelin N-terminal fragment and Fab of MORAb-009 at 2.6 Å resolution reveals an epitope encompassing multiple secondary structural elements of the mesothelin, The mesothelin fragment has a compact, right-handed superhelix structure consisting of five short helices and connecting loops. A residue essential for complex formation has been identified as Phe22, which projects its side chain into a hydrophobic niche formed on the antibody's recognition surface upon antigen-antibody contact. The overlapping binding footprints of both the monoclonal antibody and the cancer antigen CA125 explains the therapeutic effect and provides a basis for further antibody improvement.

Statistical learning confirms the diagnostic significance of the anemia panel in breast cancer

Diagnostic value of available tumor markers, such as cancer antigen CA 15-3 and carcinoembryonic antigen (CEA) in breast cancer is limited. There is an ongoing search for additional, potentially better diagnostic blood markers with improved clinical utility. The aim of this study is to evaluate performance of the approach based on routine blood tests accompanied by a statistical learning tool to the diagnosis of breast cancer. Blood was collected from total of 104 subjects which were divided into two groups: breast cancer patients and a control group that consisted of asymptomatic volunteers and patients who had benign breast lesions at the time of blood collection. Random forest statistical learning method and the external method validation have been applied to evaluate diagnostic performance of 31 routine blood tests. The applied statistical learning approach assigned the highest diagnostic importance to the anemia panel among all analyzed blood tests that also included CA 15-3. External validation has shown utility of selected statistical approach - we were able to select tests that provide a diagnostic accuracy comparable to some diagnostic tools described in literature and based on more demanding laboratory techniques, such as gene expression microarrays. Inclusion of tests for anemia significantly improves diagnostic accuracy for the breast cancer in comparison to the diagnostic accuracy of the CA 15-3 alone. Application of the random forests also enables the reduction of number of laboratory tests needed for the establishment of diagnosis. Differences in relevant test values between the cancer and control group are small but application of multiparametric statistical learning ensured diagnostic accuracy of 72.0% associated by a sensitivity of 64.7% and specificity of 84.9%.

Improved detection Of the MUC1 cancer antigen CA 15-3 by ALYGNSA fluorimmunoassay

Breast cancer is the second leading cause of cancerrelated deaths in women worldwide; a prime cancer biomarker to aid in the diagnosis, directed treatment, clinical management, and reoccurrence of this cancer is a MUC1 peptide fragment: cancer antigen 15-3 (CA 15-3). Herein, an immuno-fluorescence assay for CA 15-3 was developed; this ALYGNSA system consists of a protein biolinker (Protein G’) adsorbed onto Poly (methyl methacrylate) (PMMA). The unique interaction of Protein G’ with PMMA, a thermo-plastic polymer has been demonstrated to improve human IgG capture antibody alignment/ orientation and result in greater assay sensitivity. Indeed a previous report (HEALTH 1 325 - 329, 2009) on the shed extracellular domain of HER-2/neu revealed a 10-fold increase in sensitivity of the ALYGSNA assay over a control ELISA assay. Results from this ALYGNSA assay study revealed that a 16-fold increase in detection (≤0.94 U/mL) of CA 15-3 was found in comparison to a commercial control ELISA kit (≤15 U/mL). In conclusion, this enhanced sensitivity of the ALYGNSA assay for CA 15-3, may provide insights into the role/function of this biomarker in normal, as well as, breast cancer and other epithelial cancers.

Squamous-cell carcinoma in mature cystic teratoma of the ovary: systematic review and analysis of published data

Up to a quarter of ovarian masses originate from germ cells, and many of these are mature cystic teratomas. The secondary development of malignancy is a rare but well-known phenomenon in patients with ovarian teratomas. Squamous-cell carcinoma accounts for 80% of secondary malignant transformations of ovarian teratomas. We aimed to do an up-to-date systematic review of this rare malignant transformation. 64 suitable studies provided information on 277 patients. Squamous-cell carcinoma in mature cystic teratoma was mainly found in women aged more than 50 years, with high concentrations of squamous-cell-carcinoma antigen and cancer antigen CA125, and with ovarian tumours more than 100 mm in size. Patients with FIGO stage Ia tumours had better survival than those with more advanced disease. Complete resection together with hysterectomy, bilateral salpingo-oophorectomy and lymphadenectomy for patients with advanced disease, followed by adjuvant chemotherapy with an alkylating drug was associated with higher survival, radiotherapy was not. We make proposals for investigation and treatment of this rare disorder.

Correlation of CK‐20‐Positive Cells in Peripheral Venous Blood with Serum CEA Levels in Patients with Colorectal Carcinoma

Tumor cell dissemination appears to be an early event in tumor progression, and tumor cells can be detected in peripheral venous blood at the time of the operation. Although cytokeratin 20 (CK-20) is not specifically expressed by colorectal carcinomas, it represents a widely used marker for the detection of colorectal tumor cells. We used the combination of density centrifugation and CK-20 real-time reverse transcription polymerase chain reaction to detect CK-20-positive cells in the peripheral venous blood of 37 patients with colorectal carcinoma. Detection rates were compared to serum levels of the tumor markers carcinoembryonic antigen (CEA), carbohydrate antigen CA 19-9, and cancer antigen CA 125. The prognostic impact was assessed by the overall survival and by univariate and multivariate analysis. Overall, CK-20-positive cells in peripheral venous blood were detected in 11 of 37 (29.7%) patients. CK-20-positive patients showed a significantly higher mean serum CEA level (90.3 ng/ml) than the 4.1 ng/ml found in the CK-20-negative group (p = 0.03). CEA levels also correlated with CK-20 copy numbers. No significant correlation was observed for CA 19-9 or CA 125. CK-20-negative patients showed a trend toward better survival (p = 0.08). In the univariate analysis, CA 19-9, CEA, tumor size, lymph node status, grading, the presence of distant metastases, and resection status reached significant prognostic levels, whereas the detection of CK-20-positive cells showed only a prognostic trend (p = 0.06). Multivariate analysis failed to identify independent prognostic parameters. Here we report the correlation of CK-20-positive cells in peripheral venous blood with the serum CEA level of patients with colorectal cancer, which may represent a potential marker of the tumor load.

Synergistic effects of combined therapy using paclitaxel and [ 90Y-DOTA]776.1 on growth of OVCAR-3 ovarian carcinoma xenografts

Objective. 776.1 is a monoclonal antibody prepared against the human ovarian cancer antigen CA 125 that demonstrates preferential binding to the cell-associated form of the antigen and has shown promising results as an yttrium-90-labeled antibody in pre-clinical studies examining the effects on tumor growth in a murine xenograft model of human ovarian cancer. The purpose of the present study was to examine the effects of combined therapy with [ 90Y-DOTA]776.1 and paclitaxel compared with monotherapy with either agent on the growth of OVCAR-3 xenografts in nude mice. Methods. Mice bearing OVCAR-3 xenografts were treated with paclitaxel alone, 50 μCi or 150 μCi [ 90Y-DOTA]776.1 alone, or a combination of both treatments. Control groups were included which consisted of a nonspecific antibody, MOPC-21, labeled to a similar degree, administered as monotherapy or in combination with paclitaxel. The effects of administration of radioimmunotherapy prior to or following chemotherapy were also examined. Results. Treatment with paclitaxel and [ 90Y-DOTA]776.1 had a synergistic anti-tumor effect on the growth of OVCAR-3 xenografts. Synergy was only observed when a tumor-specific antibody was used in radioimmunotherapy. While no difference in tumor growth was observed with order of dosing, reduced toxicity was seen when paclitaxel was administered prior to radioimmunotherapy. Conclusion. The combination of radioimmunotherapy using an anti-CA 125 monoclonal antibody and chemotherapy with paclitaxel was shown to be effective in an in vivo model of ovarian cancer and may hold promise as a treatment regimen for patients with ovarian cancer.

Pharmacokinetics, Biodistribution, and Radioimmunotherapy with Monoclonal Antibody 776.1 in a Murine Model of Human Ovarian Cancer

776.1 is a murine IgG1 monoclonal antibody to the human ovarian cancer antigen CA 125 that has the unique property of having a clear preference for binding to the cell-associated form of the antigen. We have examined the tumor localization properties and efficacy of 776.1 in a subcutaneous OVCAR-3 xenograft mouse model of human ovarian cancer. Biodistribution experiments using (125)I-labeled 776.1 demonstrated a peak uptake in tumors at 72 hours postinjection, with an average of 17.7% of injected dose per gram localized to the tumor. Little uptake in other organs was observed. Further experiments using CA 125-transfected syngeneic tumors, as well as an immunoprecipitation assay using human chimeric 776.1, both clearly demonstrated that 776.1 localizes to the tumor in a CA 125-dependent manner. DOTA-776.1 (1,4,7,10-tetraazacyclododecane-N,N',N",N'" tetraacetic acid-conjugated 776.1) was labeled with (90)Y and used in efficacy studies. [(90)Y-DOTA]776.1 at a single dose of 150 microCi was able to mediate efficient reduction of tumor growth, with regression observed in a subset of animals for a period ranging from 3 to 48 days, equivalent to 3 weekly administrations of cisplatin at 6 mg/kg. No significant regression was observed in groups receiving [(90)Y-DOTA]MOPC-21 control antibody at any dose. These results suggest that 776.1 may be a promising radioimmunotherapeutic agent for the treatment of human ovarian cancer.

Characterization of serum biomarkers for detection of early stage ovarian cancer

We have previously reported the identification of three ovarian cancer biomarker panels comprised of SELDI-TOF-MS peaks representing 14 differentially expressed serum proteins for the diagnosis of ovarian cancer. Using micro-LC-MS/MS, we identified five m/z peaks as transthyretin (TTR 13.9 kDa, TTR fragment 12.9 kDa), beta-hemoglobin (Hb, 15.9 kDa), apolipoprotein AI (ApoAI, 28 kDa) and transferrin (TF, 79 kDa). Western and/or ELISA methods confirmed the differential expression of TTR, Hb, and TF, and multivariate analyses resulted in improving the detection of early stage ovarian tumors (low malignant potential and malignant; receiver operating characteristic, ROC 0.933) as compared to cancer antigen CA125 alone (ROC 0.833). Interestingly, when CA125 was included with our markers in the multivariate analysis, the ROC increased to 0.959. Furthermore, multivariate analysis with only the mucinous subtype of early stage ovarian tumors, showed our markers to greatly improve the detection of disease (ROC 0.959) as compared to CA125 alone (ROC 0.613). Interestingly, the combination of CA125 with our markers did not seem to further improve the detection of mucinous tumors (ROC 0.955). We conclude that TTR, Hb, ApoAI and TF, when combined with CA125 should significantly improve the detection of early stage ovarian cancer.

Molecular forms and microheterogeneity of the oligosaccharide chains of pregnancy-associated CA125 antigen

The cancer antigen CA125 has a very complex molecular architecture in terms of both protein backbone and oligosaccharide chains. In this study, we examined the molecular forms and microheterogeneity of oligosaccharide chains of pregnancy-associated CA125, as a first step towards gaining an insight into its possible involvement as a ligand in carbohydrate-dependent interactions. The glycobiochemical properties of CA125 may be of diagnostic and biomedical importance as specific markers of physiological and pathological conditions of early pregnancy, as well as targets in different therapeutic procedures. Pregnancy-associated CA125 was characterized by gel filtration and ion-exchange chromatography, followed by lectin-affinity chromatography with a panel of plant lectins as ligands. CA125 antigen isolated from first trimester placental extract was found to be heterogeneous in respect to molecular mass and the existence of different glyco-isoforms. Thus, elution profiles from the lectin-affinity columns demonstrated molecular subpopulations bound with low, intermediate and high affinity. Under the applied experimental conditions, CA125 bound most strongly to Triticum vulgaris agglutinin (WGA) and Ricinus communis agglutinin (RCA), but low affinity interactions occurred with the other lectins tested. The assessment of the carbohydrate composition of N- and O-glycans of pregnancy-associated CA125 was in general agreement with available data on CA125 of cancer origin. The main difference was observed in reactivity to Canavalia ensiformis agglutinin (ConA) and Phaseolus vulgaris erythroagglutinin (PHA-E) binding.

Preoperative sensitivity and specificity for early-stage ovarian cancer when combining cancer antigen CA-125II, CA 15-3, CA 72-4, and macrophage colony-stimulating factor using mixtures of multivariate normal distributions.

In CA-125-based ovarian cancer screening trials, overall specificity and screening sensitivity of ultrasound after an elevated CA-125 exceeded 99.6% and 70%, respectively, thereby yielding a positive predictive value (PPV) exceeding 10%. However, sensitivity for early-stage disease was only 40%. This study aims to increase preoperative sensitivity for early-stage ovarian cancer while maintaining the annual referral rate to ultrasound at 2% by combining information across CA-125II, CA 15-3, CA 72-4, and macrophage colony-stimulating factor (M-CSF). For direct comparisons between marker panels, all sensitivity results correspond to a 98% fixed first-line specificity (referral rate 2%). Logistic regression, classification tree, and mixture discriminant analysis (MDA) models were fit to a training data set of preoperative serum measurements (63 patients, 126 healthy controls) from one center. Estimates from the training set applied to an independent validation set (60 stage I to II patients, 98 healthy controls) from two other centers provided unbiased estimates of sensitivity. Preoperative sensitivities for early-stage disease of the optimal panels were 45% for CA-125II; 67% for CA-125II and CA 72-4; 70% for CA-125II, CA 72-4, and M-CSF; and 68% for all four markers (latter two results using MDA). Efficiently combining information on CA-125II, CA 72-4, and M-CSF significantly increased preoperative early-stage sensitivity from 45% with CA-125II alone to 70%, while maintaining 98% first-line specificity. Screening trials with these markers using MDA followed by referral to ultrasound may maintain previously high levels of specificity and PPV, while significantly increasing early-stage screening sensitivity. MDA is a useful, biologically justified method for combining biomarkers.

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.

Diagnosis of abdominal tuberculosis: Experience from 11 cases and review of the literature

To analyze the experience within our hospital and to review the literature so as to establish the best means of diagnosis of abdominal tuberculosis. The records of 11 patients (4 males, 7 females, mean age 39 years, range 18-65 years) diagnosed with abdominal tuberculosis in Harran University Hospital between January 1996 and October 2003 were analyzed retrospectively and the literature was reviewed. Ascites was present in all cases. Other common findings were weight loss (81%), weakness (81%), abdominal mass (72%), abdominal pain (72%), abdominal distension (63%), anorexia (45%) and night sweat (36%). The average hemoglobin was 8.2 g/dL and the average ESR was 50 mm/h (range 30-125). Elevated levels of cancer antigen CA-125 were determined in four patients. Abdominal ultrasound showed abnormalities in all cases: ascites in all, tuboovarian mass in five, omental thickening in 3, and enlarged lymph nodes (mesenteric, para-aortic) in 2. CT scans showed ascites in all, pelvic mass in 5, retroperitoneal lymphadenopathy in 4, mesenteric stranding in 4, omental stranding in 3, bowel wall thickening in 2 and mesenteric lymphadenopathy in 2. Only one patient had a chest radiograph suggestive of a new TB lesion. Two had a positive family history of pulmonary TB. None had acid-fast bacilli (AFB) in the sputum and the tuberculin test was positive in only two. Laparotomy was performed in 6 cases, laparoscopy in 4 and ultrasound-guided fine needle aspiration in 2. In those patients subjected to operation, the findings were multiple diffuse involvement of the visceral and parietal peritoneum, white 'miliary nodules' or plaques, enlarged lymph nodes, ascites, 'violin string' fibrinous strands, and omental thickening. Biopsy specimens showed granulomas, while ascitic fluid showed numerous lymphocytes. Both were negative for acid-fast bacilli by staining. PCR of ascitic fluid was positive for Mycobacterium tuberculosis (M. tuberculosis) in all cases. Abdominal TB should be considered in all cases with ascites. Our experience suggests that PCR of ascitic fluid obtained by ultrasound-guided fine needle aspiration is a reliable method for its diagnosis and should at least be attempted before surgical intervention.

Research of New Biomarker for Breast Cancer Using Proteomic Patterns

Purpose: Early detection and treatment of cancer is a primary focus of health care. Many serum markers are available for breast cancer, but are not good enough for screening. Cancer antigen CA 15-3 is the most widely used biomarker for breast cancer. However, CA 15-3 has low sensitivity and specificity. This study was performed to analyze the serum proteomic pattern in breast cancer patients by surface-enhanced laser desoption/ionization timeof-flight (SELDI-TOF). Methods: We screened for potential tumor biomarkers in 42 serum samples, including samples from a group of 23 breast cancer patients at different clinical stages [stage I (n=3), stage II (n=11), stage III (n=6), and stage IV (n=1)], and a control group of 19 healthy women. Diluted serum samples were applied to a C16 hydrophobic interaction chip (H4). Complex protein profiles of different groups were compared and analyzed using the Protein Chip software 2.1 (Ciphergen Biosystems). Results: There were 7 significant protein peaks in the breast cancer group and 5 in the control group. Scoring the expression of each peak, the mean score was 8.5 in the cancer group and 3.5 in the control. The results of the combination of each peak were highly sensitive (91.2%) and specific (94.7%). These proteomic patterns did not correlate with tumor stage and hormonal receptor, c-erb B2. Conclusion: In this preliminary report, we identified protein profiles that were differentiated in breast cancer patients. After proper validation, serum proteomic pattern analysis may ultimately be applied in screening breast cancer as a stand-alone or combined with current options. (Journal of Korean Breast Cancer Society 2004;7:22-26) ꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏꠏ

The cancer antigen CA125 represents a novel counter receptor for galectin-1.

CA125 is an ovarian cancer antigen whose recently elucidated primary structure suggests that CA125 is a giant mucin-like glycoprotein present on the cell surface of tumor cells. Here, we establish a functional link between CA125 and beta-galactoside-binding, cell-surface lectins, which are components of the extracellular matrix implicated in the regulation of cell adhesion, apoptosis, cell proliferation and tumor progression. On the basis of mass spectrometry and immunological analyses, we find that CA125 is a counter receptor for galectin-1, as both soluble and membrane-associated fragments of CA125 derived from HeLa cell lysates are shown to bind specifically to human galectin-1 with high efficiency. This interaction is demonstrated (1) to depend on beta-galactose-terminated, O-linked oligosaccharide chains of CA125, (2) to be preferential for galectin-1 versus galectin-3 and (3) to be regulated by the cellular background in which CA125 is expressed. Despite lacking a conventional signal peptide, a CA125 C-terminal fragment of 1148 amino acids, representing less than 10% of the full-length protein, retains the ability to integrate into secretory membranes such as the endoplasmic reticulum (ER) and the Golgi, and is targeted to the plasma membrane by conventional secretory transport. As demonstrated by a novel assay that reconstitutes non-conventional secretion of galectin-1 based on fluorescence-activated cell sorting (FACS), we find that tumor-derived HeLa cells expressing endogenous CA125 present more than ten times as much galectin-1 on their surface compared with non-tumor-derived, CA125-deficient CHO cells. Intriguingly, both the galectin-1 expression level and the cell-surface binding capacity for galectin-1 are shown to be similar in CHO and HeLa cells, suggesting that CA125 might be a factor involved in the regulation of galectin-1 export to the cell surface.

Metaplastic potential of p53 down-regulation in ovarian surface epithelial cells affected by ovulation

That ovulation is a predisposing factor in common (surface) epithelial ovarian cancer is widely recognized; however, the molecular events which underscore early-stage disease have not been elucidated. In vivo and in vitro studies using an ovine model system were designed to address a premise that oxidative distresses to DNA inflicted upon ovarian surface epithelial cells within a limited diffusion radius of the ovulatory site of follicular rupture, if gone uncorrected by p53-dependent cycle arrest/repair pathways, could yield a progenitor of tumorigenic potential. Immunofluorescence image analysis was used to quantitate the DNA damage marker 8-oxoguanine, the tumor suppressor p53, the base-excision repair polymerase β, and apoptotic internucleosomal DNA fragmentation in ovarian surface epithelial cells isolated from the perimeter of ovulated follicles. Up-regulation of p53 coincided with accretion of 8-oxoguanine lesions. Oxidative disturbances to DNA were reconciled during the consequent luteal phase (before replicative repair of the ovarian rupture wound). Production of p53 was not related to apoptosis, but rather to induction of polymerase β. Oxoguanine modifications persisted in cells affected by ovulation in which synthesis of p53 was negated in culture by an antisense oligonucleotide. Inhibition of p53 was associated with discordant cellular growth rates and expression of the cancer antigen CA-125 – a phenotype of metaplastic transformation. It is suggested that the integrity of DNA of ovarian surface epithelial cells is compromised by reactive oxidants and inflammatory mediators generated during the ovulatory process and that malfunction in a damage-recognition and(or) repair mechanism is a determinant in the etiology of ovarian metaplasia and carcinogenesis.