Common Tumor Markers Research Articles

A multiplex assay for detection of common pediatric sarcoma tumor markers

10041 Background: Molecular assays for tumor markers have become an important component in diagnosing pediatric sarcomas and small round cell tumors. We developed a multiplex assay to detect 7 common translocations associated with synovial sarcoma (SYT/SSX1, SYT/SSX2), Ewing's sarcoma (EWS/FLI1, EWS/ERG), rhabdomyosarcoma (PAX3/FKHR, PAX7/FKHR), and small round cell desmoplastic tumors (SRCDT) (EWS/WT1). Methods: 27 samples were analyzed with the multiplex assay: 13 formalin-fixed paraffin-embedded tumor samples, 5 fresh frozen tumor samples, 2 commercial RNA samples (SYT/SSX1, SYT/SSX2), 3 cell line samples (EWS/FLI, EWS/ERG, PAX3/FKHR), 1 synthetic control (PAX7/FKHR), and 3 negative controls. After RNA extraction, RT-PCR was performed using the SuperScript III One-Step RT-PCR System with Platinum Taq DNA Polymerase (Invitrogen, Carlsbad, CA). 12 primers were added to a single master mix to amplify all 7 translocations and an internal control; 1 primer from each pair was fluorescently labeled. After RT-PCR, the PCR products were separated on a genetic analyzer and the translocations identified based on the size of the PCR products and their signal intensity. Samples were also analyzed by our current assay (real-time RT-PCR or standard RT-PCR/gel electrophoresis). Results: The results of our current and multiplex assays were positive for all synthetic controls, cell lines, and commercial RNA samples and were concordant for 15/18 tumor samples: 2/7 synovial sarcoma samples were positive for SYT/SSX1 and 3/7 for SYT/SSX2; 2/6 Ewing's sarcoma samples were EWS/FLI1-positive; 1/1 SRCDT sample was EWS/WT1-positive; and 4/4 rhabdomyosarcoma samples were negative. The 3 discordant samples tested negative with our current assay; the new multiplex assay detected EWS/FLI1 in 1 of these and SYT/SSX2 in 2, in agreement with the histological classification. Conclusions: This multiplex assay can detect 7 translocations commonly found in pediatric soft tissue tumors in a single-tube reaction, with increased clinical sensitivity relative to our current methods. No significant financial relationships to disclose.

Evaluation of Matrix Metalloproteinase 7 in Plasma and Pancreatic Juice as a Biomarker for Pancreatic Cancer

Differentiating between periampullary carcinoma and chronic pancreatitis with an inflammatory mass is difficult. Consequently, 6% to 9% of pancreatic resections for suspected carcinoma are done inappropriately for chronic pancreatitis. Here, we test if matrix metalloproteinase 7 (MMP-7), a secreted protease frequently expressed in pancreatic carcinoma, can be measured in plasma, pancreatic, and duodenal juice, and if it can distinguish between periampullary carcinoma and chronic pancreatitis. Ninety-four patients who underwent pancreatic surgery for a (peri)pancreatic neoplasm (n = 63) or chronic pancreatitis (n = 31) were analyzed. Median plasma MMP-7 levels were significantly higher in carcinoma (1.95 ng/mL; interquartile range, 0.81-3.22 ng/mL) compared with chronic pancreatitis and benign disease (0.83 ng/mL; interquartile range, 0.25-1.21 ng/mL; P < 0.01). MMP-7 levels in pancreatic juice were higher, although not significantly, in carcinoma (62 ng/mg protein; interquartile range, 18-241 ng/mg protein) compared with chronic pancreatitis and benign disease (23 ng/mg protein; interquartile range, 8.5-99 ng/mg protein; P = 0.17). MMP-7 levels in duodenal juice were universally low. At an arbitrary cutoff of 1.5 ng/mL in plasma, positive and negative predictive values were 83% and 57%, respectively, values comparable to those of today's most common pancreatic tumor marker, carbohydrate antigen 19-9 (CA19-9; 83% and 53%, respectively). Positive and negative likelihood ratios for plasma MMP-7 were 3.35 and 0.52, respectively. The area under the receiver operating characteristic curve for MMP-7 was 0.73 (95% confidence interval, 0.63-0.84) and for CA19-9, 0.75 (95% confidence interval, 0.64-0.85). Combined MMP-7 and CA19-9 assessment gave a positive predictive value of 100%. Thus, plasma MMP-7 levels discriminated between patients with carcinoma and those with chronic pancreatitis or benign disease. The diagnostic accuracy of plasma MMP-7 alone is not sufficient to determine treatment strategy in patients with a periampullary mass, but combined evaluation of plasma MMP-7 with CA19-9 and other markers may be clinically useful.

Correlation study of Carcino Embryonic Antigen &amp; Cancer Antigen 15.3 in pretreated female breast cancer patients

Carcino Embryonic Antigen (CEA) and Cancer Antigen 15.3 (CA15.3) are the most common tumor markers in breast cancer patients. Measurement of circulating tumor markers is a non-invasive quantitative method. Serum levels of CEA and CA 15.3 were studied in female breast cancer patients prior to treatment. To evaluate the utility of these markers, 207 Breast carcinoma patients belonging to all the stages were considered. Healthy age matched 75 female individuals formed the control group. The serum levels of CEA and CA 15.3 were analyzed by Enzyme Linked Immunosorbent Assay (ELISA). Results were taken and compared with stages, tumor size, node and grade. The serum CA 15.3 levels were significant in all the study parameters whereas serum CEA levels showed no significant changes with any of the parameters. Measurement of serum CA 15.3 levels showed significant correlation (24.8%) with advanced stages and larger tumor sizes, whereas serum CEA levels did not show any significant correlation in breast cancer patients prior to treatment.

Increase in both CEA and CA19-9 in sera is an independent prognostic indicator in colorectal carcinoma

Carcinoembryonic antigen (CEA) and carbohydrate antigen (CA19-9) are well known to be the most common tumor markers of colorectal carcinomas. However, the significance of increase in these markers to predict the prognosis of the patients remains a problem for debate. One hundred three patients with colorectal carcinoma, who had been treated by resection and reconstruction of digestive tracts were studied. Correlation of preoperative serum value of CEA and CA19-9 with clinicopathologic features including prognosis of the patients was investigated. Preoperative elevation of both of the two markers proved to be an independent prognostic indicator, however, an elevation of only one of the two markers did not obtain a prognostic significance. Combined data of preoperative increase in CEA and CA19-9 in sera can provide a powerful and useful information to predict prognosis of patients with colorectal carcinoma.

Circulating hTERT mRNA as a tumor marker in cholangiocarcinoma patients

To investigate human telomerase reverse transcriptase (hTERT) mRNA in the serum of cholangiocarcinoma patients. The serum of thirty three cholangiocarcinoma patients, forty one benign biliary tract disease patients and ten healthy volunteers were collected and analyzed for the expression of hTERT mRNA by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). We then examined the correlation between values of serum hTERT mRNA and the pathological staging of cholangiocarcinoma. hTERT mRNA was detected in 28 of 33 (84.85%) of serum obtained from cholangiocarcinoma patients and 9 of 41 (21.9%) of serum obtained from benign biliary tract disease patients. hTERT mRNA was not detected in any serum obtained from healthy volunteers. on the other hand the common tumor marker, CA19-9 was detected in 20 of 33 (60.6%) of serum obtained from cholangiocarcinoma patients and 8 of 41 (19.5%) of serum obtained from benign biliary tract disease patients. However, no correlation was found between the present of serum hTERT mRNA and tumor staging. These results indicate that the detection of circulating hTERT mRNA was identified in almost all cholangiocarcinoma patients. It offers a novel tumor marker, which can be used as a complementary study for diagnosis of cholangiocarcinoma.

Diagnostic Value of CYFRA 21-1, CEA, CA 19-9, CA 15-3, and CA 125 Assays in Pleural Effusions: Analysis of 116 Cases and Review of the Literature

Levels of tumor markers in pleural effusions may help to establish the diagnosis of pleural malignancy, but the precise diagnostic value of each marker remains unclear. The aim of this study was to assess the diagnostic value of five common pleural fluid tumor markers, carcinoembryonic antigen (CEA), cytokeratin fragment (CYFRA) 21-1, cancer antigen (CA) 15-3, CA 19-9, and CA 125, and to review the literature from the past 15 years. Pleural fluid samples were collected prospectively from 116 patients and assayed for CEA, CYFRA 21-1, CA 15-3, CA 19-9, and CA 125 levels. A MEDLINE search of the English-language literature from the past 15 years was also done. Effusions were classified as benign or malignant on the basis of their definitive pathologic or cytologic diagnoses. The levels of all pleural tumor markers were statistically significantly higher in the malignant group than in the benign group. The marker with the highest accuracy was CEA (85.3%); CA 15-3, CYFRA 21-1, and CA 19-9 had similar accuracies (75.2%, 72.4%, and 71.5%, respectively), and CA 125 had the lowest accuracy (40.5%). On univariate analysis, tumor-marker combinations did not result in a greater accuracy than that of CEA alone. On multivariate logistic regression, CA 15-3 and CYFRA 21-1 were significant predictors of malignancy. Among the nine reports in the literature comparing 11 different tumor markers, CEA, CA 15-3, and CYFRA 21-1 yielded the best results. We conclude that pleural fluid analysis should include CEA for the diagnosis of malignancy. CA 15-3 and CYFRA 21-1 may serve as alternative options.

Clinical Significance of Osteopontin in Esophageal Squamous Cell Carcinoma: Comparison with Common Tumor Markers

Objective: Osteopontin (OPN) is a secreted integrin-binding glycophosphoprotein that may have a role in head and neck squamous cell carcinoma (SCC). To evaluate the clinical significance of OPN in esophageal squamous cell carcinoma (ESCC), we compared plasma OPN levels with those of common tumor markers. Methods: Preoperative plasma OPN levels were measured by enzyme immunoassay in 103 ESCC patients. Serum SCC antigen, Cyfra 21-1, and carcinoembryonic antigen (CEA) levels were also measured routinely at admission by radioimmunoassay. Results: Plasma OPN levels ranged from 82.8 to 1,980 ng/ml. High OPN level was associated with lymph node metastasis (p = 0.05), but not with tumor histology or depth of invasion. The overall survival of the patients with high OPN levels was worse than that of those with low OPN levels (p = 0.02). SCC antigen and Cyfra 21-1 levels were associated with the depth of tumor invasion, the tumor diameter, lymph node metastasis, and the overall survival, but CEA was not associated with these clinicopathological factors. Combined evaluation of OPN plus Cyfra 21-1 or OPN plus SCC antigen was useful as an independent prognostic indicator. Conclusion: Measurement of the plasma OPN level, as well as serum SCC antigen and Cyfra 21-1, may help to predict the progression of ESCC.

Is it reasonable to add preoperative serum level of CEA and CA19-9 to staging for colorectal cancer?

Carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9) are the most common tumor markers for colorectal cancer. The aim of this study was to evaluate the possibility of adding them into the current staging system by analyzing their prognostic significance. The study population was patients (n = 574, 67.1 +/- 11.3 years old, 397 males) who received potentially curative resection of colorectal adenocarcinoma (stage I-III) between January 1994 and August 2002, including preoperative measurements of CEA and CA19-9. Clinicopathological characteristics and associated follow-up data were retrospectively collected by reviewing available medical charts. CEA higher or equal to 5 ng/ml was defined as abnormal (CEA+). The CA19-9 level was set at 37 U/ml (CA19-9+). Patients were further divided into four groups (1, 2, 3, 4) according to the results of these two markers (CEA/CA19-9: -/-, -/+, +/-, and +/+). Survival was analyzed for AJCC staging, CEA (+) versus (-), CA19-9 (+) versus (-), and four groups. CEA and CA19-9 survival curves were not significantly different. However, the combined use of the two markers revealed a significant survival benefit (P = 0.035) of group 1 ("-" for both markers) over 4 ("+" for both) in stage II. Patients with an elevated level of both CEA and CA19-9 in stage II of colorectal cancer have a significantly poorer prognosis than those with normal levels of these markers. We recommend adding both CEA and CA19-9 to the current staging system.

Combined use of established and novel tumour markers in the diagnosis of head and neck squamous cell carcinoma

Serum SCC, CYFRA 21-1, and CEA are the common tumour markers for head and neck squamous cell carcinoma (HNSCC), although diagnostic sensitivity should be yet improved, especially at early stages. In the present study, we have reported the diagnostic value of two novel serum tumour markers in HNSCC: alpha-L-fucosidase (AFU) activity, and total sialic acid concentration adjusted by total protein concentration (TSA/TP). Using the cut-off 4.0 U/ml, AFU showed a sensitivity of 55% with specificity levels of 91%, 85% and 50% to discriminate HNSCC patients from healthy donors, drinking and smoking subjects, and patients with benign diseases, respectively. Furthermore, AFU showed the best sensitivity (71%) in the detection of patients with premalign lesions. Using the cut-off 12.0 ng/mg, TSA/TP showed the best sensitivity levels (63%) in the diagnosis of HNSCC with specificity levels of 94%, 50% and 90%, regarding healthy donors, drinking and smoking subjects, and patients with benign diseases, respectively. It was of special interest that sensitivity in the diagnosis of HNSCC at non-disseminated stages was improved when using combinations of AFU+CYFRA or TSA/TP+CYFRA, up to 86% or 71% in TNM I, 60% or 80% in TNM II, and 80% or 60% in TNM III, respectively.

A novel tumour marker RCAS1 in a case of extramammary Paget's disease

A 66-year-old male, whose primary skin lesion in extramammary Paget's disease had been surgically resected 4 years previously, was hospitalized with liver metastases. Hepatic arterial infusion chemotherapy was carried out and the tumours clearly reduced in size. Serum levels of some common tumour markers were not elevated, even prior to therapy. We measured serum levels of a novel tumour-associated antigen, RCAS1, because its expression was detected in the tumour cells. The patient's serum RCAS1 level was elevated (22.0 U/mL) before therapy and fell during (10.5 U/mL) and after (5.0 U/mL) therapy. Therefore, serum RCAS1 levels may be valuable as a potential biomarker for monitoring therapeutic efficacy against Paget's disease.

COST-EFFECTIVENESS OF A MODIFIED CARE PROTOCOL SUBSTITUTING BLADDER TUMOR MARKERS FOR CYSTOSCOPY FOR THE FOLLOWUP OF PATIENTS WITH TRANSITIONAL CELL CARCINOMA OF THE BLADDER: A DECISION ANALYTICAL APPROACH

Decision analysis models were established to determine the cost-effectiveness of a plan alternating a bladder tumor marker with cystoscopy and cytology at 3-months intervals or modified care versus followup cystoscopy and cytology every 3 months or standard care. We performed a comprehensive literature review for bladder tumor markers using 1966 to current MEDLINE data and other search engines. Statistical performance data on tumor markers were calculated by hierarchical Bayes meta-analysis with 95% confidence intervals used for the model. Other basic assumptions included cost data on tumor markers, cytology, cystoscopy, transurethral bladder resection and cystectomy as well as recurrence and progression rates from the literature. Decision trees were built with computer software using a 12 and a 24-month model with linear recurrence and progression rate assumptions. Overall specificity for common tumor markers was 73% to 90% and sensitivity was 49% to 77%. As expected, sensitivity increased for higher disease grades and stages. The modified care protocol was more cost-effective than standard care for the 1 and 2-year followups. On 1-year 1-way sensitivity analysis for cost only tumor marker cost, recurrence and progression rates achieved a threshold at which the modified care protocol was as or more expensive as standard care. Modified care was more cost-effective at a tumor marker cost of less than $264. On 2-way sensitivity analysis there was no significant impact of parameters at a wide range of tumor marker costs, recurrence and progression rates. Using a modified followup protocol in transitional cell carcinoma cases with an initial tumor occurrence using a urine based tumor marker alternating with cystoscopy and/or cytology is cost-effective for a wide range of marker sensitivities, specificities and costs, cystoscopy and/or cytology cost, a 20% to 80% yearly recurrence and a 2% to 40% yearly progression rate. This protocol should be evaluated in a prospective randomized trial to determine whether the financial and emotional benefits outweigh any potential drawbacks.

Serum CYFRA 21-1 in cervical cancer patients treated with radiation therapy.

A fragment of cytokeratin 19, referred to as CYFRA 21-1, is abundant in the serum of many patients with malignant tumors and is recognized as one of the established tumor markers, especially for non-small-cell lung cancer. In this study, the clinical usefulness of CYFRA 21-1 was investigated in cervical cancer patients treated with radiation therapy with reference to squamous-cell-carcinoma-related antigen (SCC-Ag), a common tumor marker of cervical squamous cell carcinoma. The serum levels of CYFRA 21-1 and SCC-Ag of 50 patients with squamous cell carcinoma of the uterine cervix were measured before and after radiation therapy. CYFRA 21-1 was positive in 52% of the patients. The incidence increased with the stage of the cancer, and post-treatment increases were a sign of disease progression. During radiation, serum levels of CYFRA 21-1 decreased significantly and reflected the radiation effect well. In addition, CYFRA 21-1 was negative in all patients without distant metastasis at the end of radiation therapy. Compared with SCC-Ag, patients were less often positive for CYFRA 21-1, but there was a statistically positive correlation between the two markers (correlation matrix=0.69). CYFRA 21-1 can be used in monitoring the outcome of patients with squamous cell carcinoma of the uterine cervix. It may be particularly useful for patients without SCC-Ag.

TUMOR MARKERS: An Update

Cancer remains the second leading cause of death in developed countries, and the incidence of certain tumors is increasing despite emphasis on prevention and screening. Tumor markers are biologic or biochemical substances that are produced by tumor cells and then secreted into the circulation in detectable amounts. This article covers some of the more common tumor markers currently being utilized for diagnostic and prognostic purposes.

Clinical evaluation of common serum tumor markers (TM) in patients (PTS) with bladder carcinoma (BC)

Clinical evaluation of common serum tumor markers (TM) in patients (PTS) with bladder carcinoma (BC)

Molecular marker test standardization.

A review of the status of standardization of laboratory tests of particular interest to oncologists is presented. Currently, relatively few of these tests are standardized; as a result, interlaboratory and interinstitutional comparison of data is problematic. In 1992, additional interlaboratory studies of common tumor markers will be initiated by the College of American Pathologists. The National Committee for Clinical Laboratory Standards also has begun to develop standard methods and guidelines for these important tests.

Aggressive “Breast-like” Adenocarcinoma of Vulva

A case o f vulvar adenocarcinoma seen in a 42-year-old woman is reported. The tumor involved the right labium majus and diffusely spread into the surrounding soft tissues as well as the inguinal lymph nodes and was histologically composed o f nests, cords, and tubular formations recalling an aggressive duct carcinoma o f breast. Likewise, tumor cells exhibited positivity for common breast tumor markers, such as epithelial membrane antigen, carcinoembryonic antigen, and glandular keratins. The possible origin o f the neoplasm from mammary ecto pic tissue in vulva or from vulvar skin adnexa is briefly discussed.

The Danish Breast Cancer Group (DBCG) biochemical tumor marker program.

One hundred and thirty-three high-risk breast cancer patients entered the DBCG (Danish Breast Cancer Cooperative Group) tumor marker program between May 1978 and June 1980. Eleven were excluded from the DBCG protocol and 23 left the program prematurely. The remaining patients were seen at the clinical laboratory once a month during the first year and every third month during the second and third year following mastectomy. Blood and urine specimens were collected and plasma, serum and urine specimens stored at -80 degrees C. The purposes of the program were 1) to collect a biological material which may later be used to assess the usefulness of biochemical markers for the prediction of recurrence and 2) to develop statistical time series models suitable for this assessment. Such a model is presented and its application for the assessment of the usefulness of 14 common tumor markers reviewed. Only P-CEA proved to be of any value but its usefulness was marginal.

Identification of tumor-associated antigens and their purification from cyst fluids of ovarian epithelial neoplasms

Based on several earlier reports demonstrating the existence of ovarian tumor-associated antigens in the cyst fluids of adenocarcinoma, we attempted to confirm these results, purify the tumor-associated antigens, and characterize the molecular species. Several antisera were produced in rabbits with clarified whole mucinous cystadenocarcinoma fluids. Unadsorbed antisera demonstrated very complex immunodiffusion patterns. Following adsorption with normal human serum and plasma and extracts of normal cervix and ovary strong cyst-associated reactivity remained. Further adsorption with A+ and B+ erythrocytes had no effect on antisera activity. A major cyst fluid antigen was purified from mucinous cysts by ammonium sulfate fractionation (0.20–0.60 saturation), DEAE-agarose chromatography, Sephadex G-200 gel filtration, and Bio-Gel A-5m agarose gel filtration. Polyacrylamide gel electrophoresis gave a single prominent band with minor contaminants. Approximately 70% of purified and radiolabeled antigen was precipitable by antisera, and 80% precipitated in either 10% trichloroacetic acid or 0.6 M perchloric acid. Tissue distribution was assessed by immunodiffusion analysis and showed strong association with mucinous tumors (benign and malignant) of the ovary. Purified antigen showed no reactivity by immunodiffusion analyses against antisera to other common tumor markers including carcinoembryonic antigen, alphafetoprotein, ferritin, and pregnancy-associated glycoproteins.