Characterization of new human pancreatic cancer cell lines which propagate in a protein-free chemically defined medium

Many human cancer cell lines which have been maintained in fetal bovine serum (FBS)-supplemented medium produce and secrete many substances such as transferrin, alpha 1-antitrypsin, alpha 2-macroglobulin, alkaline phosphatase, gamma-glutamyltranspeptidase, creatine kinase, carcinoembryonic antigen, alpha-fetoprotein, carbohydrate antigen 19/9, and cytokines including colony-stimulating factors and transforming growth factor, and further they may produce small amounts of unknown substances. Usually, small amounts of substances have to be concentrated as highly as possible for detection, but FBS interferes with this procedure. A protein-free culture system is an ideal method for detecting small quantities of substances which originate from cancer cells without interference by FBS. However, we were concerned that protein-free culture may interrupt the production of the substances which have been produced in FBS-supplemented medium. In this study, we investigated the productibility of 46 kinds of well-known substances in ten newly established cell lines derived from human pancreatic cancer. These cell lines were propagated in a protein-free non-FBS-supplemented medium. Of the ten cases, one cell line alone that was derived from acinal cell carcinoma propagated as a semisuspension; on the other hand, nine cell lines that were derived from ductal cell carcinoma propagated as monolayers without piling up. This method prolongs the doubling time, which is not affected by the addition of FBS. The spent media of these cell lines were collected aseptically after the removal of cell debris and concentrated by ultrafiltration using a Pericon cassette followed by lyophilization. Using 46 kinds of available antibodies, we investigated whether or not the substances which react to these antibodies could be detected in the spent media and in the cells by enzyme-linked immunosorbent assay, Western blot analysis, and immunocytochemistry. Among these cell lines, HPC-Y11 produced and secreted the most kinds of substances, and the production of those substances was lowest in HPC-Y0. In conclusion, our protein-free culture system can be available in every laboratory, since this is not only an economical method, but also an effective method for the saving of purification procedures. Moreover, this is a most suitable method for surveying unknown substances derived from cancer cell lines.     

Flow cytometric identification of cell surface markers on cultured human colonic cell lines using monoclonal antibodies

Background. The expression of tumor-associated cell surface antigens is a reflection of the state of cell differentiation of tumor cells in culture. Method. Monoclonal antibodies (MoAbs) against the tumor-associated antigens carcinoembryonic antigen (CEA) and CA19-9 and the extracellular matrix protein CD44 were used to label the cell surface of human colonic cells in culture. The binding of each antibody to its respective antigen was measured by fluorescence-activated flow cytometry and expressed as a percentage of positive cells. Results. The human colon adenocarcinoma cell (HCAC) line, LS-180, showed strong binding with CEA (81%), CA 19-9 (87%), and CD44 (83%). LS-174t cells, a trypsinized variant of LS-180 cells, showed less binding with CEA (66%) and CA 19-9 (49%), but no binding with CD44. With cells from HCAC line HT-29, antigen expression was highly variable for CEA (13% ± 18) and CD44 (31% ± 35) but was consistently positive for CA19-9 (33% ± 13). The expression of CEA in the Caco-2 cell line was weak (24%), whereas there was no expression of CA19-9 and CD44. Normal human colon fibroblast cells (CCD-18Co) did not recognize the monoclonal antibodies to CEA or CA 19-9, but were strongly positive with the CD44 antibody (97%). Conclusions. These results support the concept that the expression of the tumor associated markers CEA and CA19-9 and the cell surface marker CD44 on human colonic cell lines varies with the degree of cellular differentiation. Carcinoembryonic antigen and/or CA19-9 were expressed in all four human colon adenocarcinoma cell lines, but not in the normal colon fibroblast cells (CCD-18Co). Using these two MoAbs appeared to be a more reliable measure of the state of differentiation of human colon adenocarcinoma cells. Cancer 1995; 75:195–200.     

Tumorigenicity in athymic mice of the human colon carcinoma cell line SW1116 expressing the tumor-associated antigenic determinant CA 19-9

The human colorectal carcinoma cell line SW1116 under optimal growth conditions synthesized and shed antigens bearing the monoclonal antibody-defined carbohydrate determinant CA 19-9. Antigen expressing CA 19-9 in cell culture supernatant was quantitated by an immunoradiometric assay for CA 19-9. Injection of SW1116 cells s.c. into athymic BALB/c mice resulted in the growth of moderately differentiated tumors possessing a distinct morphological resemblance to a typical adenocarcinoma of the colon. Intervals to tumor appearance were dependent on inoculum dose, but 95% of mice at both 5 X 10(6) and 10(7) cells/mouse developed tumors within 14 to 21 days. CA 19-9 antigen was detected in the sera of all nude mice with SW1116 tumors, and antigen concentration correlated (r = 0.77) with tumor volume throughout the 9-week study. The half-life of this antigen in serum following tumor excision from nude mice was 6.5 +/- 1.5 (S.D.) hr. Carcinoembryonic antigen was also detected in serum from mice bearing SW1116 tumors by an immunoradiometric assay for carcinoembryonic antigen, but its concentration correlated (r = 0.86) with tumor volume for only the first 4 weeks of tumor growth. Significant levels of endogenous immunoglobulin G1 and immunoglobulin G3 antibodies to CA 19-9 antigen were found in the serum of nude mice with SW1116 tumors by radioimmunodiffusion, but no apparent relationship between antibody titer and tumor growth or CA 19-9 antigen level in serum was evident. This tumor model may be useful in devising radioimmunodetection and immunotherapeutic strategies for primary and metastatic human colon carcinomas.     

Establishment and characterization of a new cell line from primary human breast carcinoma

Summary A new cell line (BRC-230) was established from surgical material of primary ductal infiltrating breast carcinoma. The epithelial nature of this cell line was confirmed by ultrastructural analysis and demonstrated the retention of structural properties characteristic of the original tumor. The BRC-230 cell line induced tumor in athymic Cr1:nu/nu(CD-1)BR nude mice, it possessed an abnormal karyotype with a modal chromosome number between 60–61 with eight recurrent marker chromosomes, and it presented a doubling time of 30.5 hr. Scatchard analysis demonstrated that both primary tumor and BRC-230 cells were estrogen and progesterone receptor negative.Immunoenzymatic and radioimmunoassays showed a production of marker antigens (CEA, TPA, CA125, CA15-3, CA19-9) which was similar in the patient's serum and BRC-230 cells. Thein vitro drug sensitivity assay of the cell line and of the parental tumor tissue showed overlapping results to all tested antiblastic drugs. BRC-230 cells were resistant to 4-Idroperoxy-cyclophosphamide, Idarubicinol, Mitoxantrone, Etoposide, 4Epidoxorubicin, and Doxorubicin, showing a multiple drug resistance phenotype. Amplification or rearrangement of Her-2neu, Ha-ras, and C-myc genes was observed neither in the original tumor nor in BRC-230 cells; the mdr-1 gene was also present in a single copy.We conclude from these studies that the BRC-230 cell line maintains the same characteristics as the original tumor and may provide us with a good model to studyin vitro the biology of drug resistance of breast cancer.     

Normal bronchial mucus contains high levels of cancer-associated antigens, CA125, CA19-9, and carcinoembryonic antigen

The presence of cancer-associated antigens CA125, CA19-9, and carcinoembryonic antigen (CEA) in apparently normal respiratory system was demonstrated histochemically and immunochemically. Epithelial cells lining central airways (trachea, bronchi, and bronchioli) and respiratory glands were specifically stained by antibodies recognizing CA125, CA19-9, and CEA. Most, if not all, bronchial mucus obtained from patients without pulmonary diseases during general anesthesia contained remarkably high levels of CA125, CA19-9, and CEA ranging from 190 to 41,000 U/ml (594-4803 U/mg protein), 210 to 95,000 U/ml (294-197,917 U/mg protein), and 6 to 940 ng/ml (14-209 ng/mg protein), respectively, whereas serum antigen levels were normal in all cases examined. These results suggest that CA125, CA19-9, and CEA are synthesized and secreted by normal epithelial cells of central airways and/or respiratory glands and that these substances are not specific indicators of abnormal cellular activity.     

Growth and differentiation of a murine interleukin-3-producing myelomonocytic leukemia cell line in a protein-free chemically defined medium.

We established the continuous growth of WEHI-3B D+ cells in protein-free chemically defined F-12 medium by stepwise decreases in the concentration of fetal calf serum. This cell line, designated as WEHI-3B-Y1, has now been propagated in protein-free F-12 medium for 3 years. The population-doubling time of the cells in culture is about 24 hr. WEHI-3B-Y1 cells are immature undifferentiated cells which show positive staining for naphthol ASD chloroacetate esterase and alpha-naphthyl butyrate esterase and spontaneously exhibit a low level of differentiation to mature granulocytes and macrophages. Medium conditioned by WEHI-3B-Y1 cells stimulated the proliferation of an interleukin-3 (IL-3)-dependent FDCP-2 cell line. This conditioned medium was shown to have erythroid burst-promoting activity when assayed using normal murine bone marrow. The colony formation of WEHI-3B-Y1 cells in semi-solid agar culture was not stimulated by purified recombinant human granulocyte colony-stimulating factor (rhG-CSF). However, in the presence of human transferrin, rhG-CSF enhanced the number of colonies of WEHI-3B-Y1 cells but did not induce their differentiation. These results suggest that WEHI-3B-Y1 cells cultured in protein-free medium produced murine IL-3. In addition, human G-CSF enhanced the clonal growth but did not induce the differentiation of WEHI-3B-Y1 cells cultured in serum-free medium.     

Characterization of a human ovarian carcinoma cell line, OTN 14, derived from a mucinous cystadenocarcinoma

A human ovarian carcinoma cell line, OTN 14, has been established from malignant ascitic fluid of a patient with a well-differentiated mucinous cystadenocarcinoma of the left ovary. The cell line has been maintained in vitro for 6 months through 23 passages, growing in monolayers as well as in 3-dimensional clusters, with a population doubling time of 28 1/2 hr. The number of chromosomes per cell varied from 67 to 88, with a modal number of 86. Two characteristic marker chromosomes were recognized, consisting of partially deleted chromosome I. With a DNA index of 1.934 the tumour cell line was near tetraploid. The epithelial character of the OTN 14 cells was confirmed by a positive immunofluorescence reaction with monoclonal antibodies (MAbs) against different keratins, and when (immuno)electron microscopy was used, keratin filaments and small junctional complexes were observed. Vimentin was also expressed in these cells, while desmin was not detected. Cultured tumour cells reacted (weakly) positive with MAb OV-TL 3 as a marker for ovarian carcinomas, while reactivity with the anti-ovarian carcinoma MAb OC 125 was limited to a few cells, not permitting the detection of shed CA 125 antigen in the culture supernatant. Cells stained heterogeneously positive for CEA marker BW 431/31, the presence of which was confirmed by detection of CEA shed into the culture medium. The cell line released estradiol at a concentration of 130,000 pmol/L in the culture medium, while no progesterone or dehydroepiandrosterone sulphate were found. Electron microscopical evidence for steroid production was suggested in some cells showing "dense-core" vesicles near the Golgi areas. The OTN 14 tumour cells formed poorly differentiated tumour nodules in nude mice, and metastatic cells were also found in blood capillaries. Cell types with mucinous as well as endocrine characteristics were found.     

Carcinoembryonic antigen and carbohydrate antigen 19-9 levels of peripheral and draining venous blood in colorectal cancer patients. Correlation with histopathologic and immunohistochemical variables

Correlation between carcinoembryonic antigen (CEA) and carbohydrate antigen (CA) 19-9 levels of peripheral and draining venous blood, and 11 histopathologic and immunohistochemical variables was examined in 83 patients with colorectal cancer. CEA levels of draining blood (mean 34.5 ng/ml and positive rate greater than 5 ng/ml, 60.2%) were significantly higher than those (13.0 ng/ml and 28.9%) of peripheral blood. However, CA19-9 levels (mean 576.1 U/ml and positive rate greater than 37 U/ml, 29.5%) of draining blood were not different from those (568.0 U/ml and 29.5%) of peripheral blood. Immunohistochemically, CEA was observed in all of the 83 specimens and distributed in most of all cancer cells, whereas CA19-9 was found in 52 (62.5%) of the 83 specimens and sporadically distributed in some parts of cancer lesions in general. Elevation of CEA levels in draining and peripheral blood was most highly correlated with venous invasion, although the levels were related to four other histopathologic variables including liver metastasis, invasive layer of colorectal wall, lymphatic invasion, and Dukes' classification. Significant correlation between the CEA localized pattern of cancer cells was not found. Patients with CA19-9 nonlocalized cancer showed no elevation of the antigen levels in both peripheral and draining blood. The elevation of CA19-9 levels in peripheral blood of patients with CA19-9 localized cancer was most highly associated with lymphatic invasion, although the levels were correlated with five other variables consisting of liver metastasis, tumor differentiation, invasive layer of colorectal wall, venous invasion, and Dukes' classification out of 11 histopathologic and immunohistochemical variables. CEA levels of draining blood rose from 18.2 ng/ml and 40.3% to 30.1 ng/ml and 72.6%, respectively, after operative stimuli to cancer lesions, whereas the change of CA19-9 levels in draining blood of patients with CA19-9 localized cancer was not found during the time of operation. These results suggest that CEA may be drained mainly by the hematogenous portal system by the draining vein from the cancer cells in the invasive veins and that CA19-9 may be drained by the thoracic duct of the lymphatic system. It is also suggested that the CEA and CA19-9 elevation-relating variables may secondarily affect the CEA and CA19-9 elevation in the blood in association with the venous and lymphatic invasion of cancer lesions, respectively.     

Measurement of a monoclonal-antibody-defined antigen (CA19-9) in the sera of patients with malignant and nonmalignant diseases. Comparison with carcinoembryonic antigen

Immunoradiometric assay (IRMA) using monoclonal antibody for colon cancer cell surface antigen (CA19-9) was compared with carcinoembryonic antigen (CEA) with regard to sensitivity and specificity in 730 patients. In the 341 patients who had no evidence of malignant disease, CA19-9 levels ranged between less than 1.5 to 49 U/ml. Specificity of CA19-9 at a cutoff of 20 U/ml was similar to that of CEA at a cutoff of 5.0 ng/ml; CA19-9 was more sensitive than CEA in pancreatic cancer, whereas CEA was more sensitive than CA19-9 in breast, colon, and gastric cancer. Of 17 patients with pancreatic cancer, 13 had elevated levels of CA19-9 (sensitivity, 76%), whereas only 8 had elevated levels of CEA (sensitivity, 47%) and 15 had elevated levels of either CEA or CA19-9 (sensitivity, 88%). These findings suggest that, like CEA, CA19-9 is detectable in nonmalignant diseases and is not specific for gastrointestinal tumors, and has higher sensitivity than CEA only in pancreatic cancer. However, further prospective studies are required to verify its value in the diagnosis and management of pancreatic cancer.     

Clinical evaluation of the simultaneous determination of tumor markers in fluid and serum and their ratio in the differential diagnosis of serous effusions.

We evaluated the diagnostic utility of simultaneous determination of 5 tumor markers, CEA, CA 125, CA 15-3, CA 19-9 and cytokeratin 19 (CYFRA 21-1), in fluid and serum from 101 patients, 52 with pleural effusion (22 malignant) and 49 patients with ascites (14 malignant). Tumor marker concentrations in fluid from patients with malignant effusions were significantly higher than those obtained in benign fluids or serum. However, there are two types of tumor markers: those released/secreted by normal mesothelia such as CA 125 and cytokeratin 19 (higher levels in benign fluids than in serum) and non-released/secreted tumor markers (low concentrations in benign fluids) such as CEA, CA 19-9 and CA 15-3. The fluid/serum (F/S) ratio showed better sensitivity with maximum specificity than a single determination in fluid for CEA, CA 15-3 and CA 19-9, but not for CA 125 and CYFRA. The combination of a F/S ratio greater than 1.2 and a cut-off of 5 ng/ml for CEA, 30 U/ml for CA 15-3 and 37 U/ml for CA 19-9 showed sensitivities of 58, 57 and 44%, respectively, and a specificity of 100%, with a combined sensitivity of 82% for overall effusions and 79% for fluids with negative cytology with a specificity of 100%. In conclusion, the use of the F/S ratio in nonsecreted tumor markers such as CEA, CA 19-9 and CA 15-3 improve the sensitivity and specificity and allow standardization of the cut-off.     

Establishment and characterization of a human pancreatic cancer cell line (SUIT-2) producing carcinoembryonic antigen and carbohydrate antigen 19-9

A new tumor cell line (SUIT-2) derived from a metastatic liver tumor of human pancreatic carcinoma has been established in tissue culture and in nude mice, and maintained for over five years. In tissue culture, the cells grew in a monolayered sheet with a population doubling time of about 38.2 hr, and floated or piled up to form small buds above the monolayered surface in relatively confluent cultures. Chromosome counts ranged from 34 to 176 with a modal number of 45. Subcutaneous injection of cultured cells into nude mice resulted in tumor formation, histopathologically closely resembling the original neoplasm which had been classified as moderately differentiated tubular adenocarcinoma. Electron microscopic observation of the neoplastic cells revealed a characteristic pancreatic ductal epithelium. SUIT-2 cell line produces and releases at least two tumor markers, carcinoembryonic antigen and carbohydrate antigen 19-9, propagates even in serum-free medium, and metastasizes to the regional lymph nodes in nude mice xenografts.     

gamma-Glutamyltranspeptidase activity in a human pancreatic cancer cell line (HPC-Y1) in serum-free chemically defined medium

gamma-Glutamyltranspeptidase (gamma-GTP) activity was found in a human pancreatic cancer cell line, HPC-Y1, cultivated in a serum-free chemically defined medium. The gamma-GTP stained in the cytoplasms as fine granules and was produced constantly in a protein-free chemically defined medium. The detergent- or protease-solubilized gamma-GTP of HPC-Y1 cells in serum-free medium was compared with the gamma-GTPs extracted from HPC-Y1 cells in serum-containing medium, human pancreatic carcinoma and normal human pancreas. Their molecular weights, electrophoretic mobilities, affinity to Concanavalin A-Sepharose and isoelectric points were almost identical. No cancer specific properties in the gamma-GTP derived from human pancreatic carcinoma cell line were found by these analyses. However, the serum-free spent medium of HPC-Y1 cells was useful for purifying the gamma-GTP secreted from the human pancreatic carcinoma cell line, since it is not necessary to separate the contaminated serum components that are usually added for cell cultures and the extraction procedures could induce minor structural change and/or artificial modification of gamma-GTP.     

FH检测在直肠恶性肿瘤早期筛查中的诊断价值

 目的 探讨直肠上皮细胞稳定性FH检测在直肠恶性肿瘤早期筛查中的诊断价值。方法 300例被检者均行直肠上皮细胞稳定性FH、CEA和CA19-9检测。以病理学诊断结果为金标准，辅以电子结肠镜检查，分析上皮细胞稳定性FH、CEA和CA19-9检测对直肠恶性肿瘤筛查的敏感度、特异性、假阴性率（漏诊率）、假阳性率（误诊率）、阴性似然比、阳性似然比、诊断指数、约登指数、粗符合率、阴性预测值、阳性预测值等指标。结果 经上皮细胞稳定性FH检测直肠恶性肿瘤患者阳性率高于CEA和CA19-9检测（P=0.000）。40例直肠恶性肿瘤患者中上皮细胞稳定性FH检测出阳性患者38例（95%），CEA检测出阳性患者23例（57.50%），CA19-9检测出阳性患者10例（25.00%）。上皮细胞稳定性FH检测的敏感度、阳性似然比、诊断指数、约登指数、阴性预测值、阳性预测值均高于CEA和CA19-9检测，假阴性率和阴性似然比均低于CEA和CA19-9检测。结论 上皮细胞稳定性FH检测在直肠恶性肿瘤筛查中检测结果有较高的敏感度、阳性似然比、诊断指数、约登指数、阴性预测值、阳性预测值，较低的假阴性率和阴性似然比。     

CA19-9 epitope a possible marker for MUC-1/Y protein

It has been reported that MUC-1 molecules devoid of the tandem repeat region (MUC-1/Y) are detected preferentially in carcinoma cells and are associated with their progression. However, its clinical significance is still unknown. We constructed a mouse colon adenocarcinoma cell line (MC-38) transduced with either MUC-1/Y cDNA defecting the tandem repeat region (Y-MC-38) or MUC-1/R cDNA containing ten tandem repeats (R-MC-38). RT-PCR of mRNAs derived from Y-MC-38 cells using the specific primers to MUC-1/Y mRNAs, proved the existence of 600 bp RT-PCR products generated only from MUC-1/Y mRNAs. DF3 and CA19-9 epitopes out of the MUC-1-related tumor-associated antigens, have been reported to be involved in the prognosis of cancer patients. We examined the expression of DF3 and CA19-9 epitopes on Y-MC-38 and R-MC-38 cells. Fluorescence-activated cell sorting (FACS) analysis of R-MC-38 and Y-MC-38 cells using two monoclonal antibodies against DF3 (mAb DF3) and CA19-9 (mAb CA19-9) epitopes revealed that R-MC-38 cells expressed DF3 but not CA19-9 [DF3(+)CA19-9(-)], while Y-MC-38 cells expressed CA19-9 but not DF3 [DF3(-)CA19-9(+)]. On Western blot, a 40 kDa protein product was recognized by mAb CA19-9 but not by mAb DF3 on cell lysates of Y-MC-38 cells, whereas a 70 kDa protein product was recognized by mAb DF3 but not by mAb CA19-9 on the cell lysates of R-MC-38. Further, we analyzed the expression of MUC-1/Y mRNAs by RT-PCR on various human cancer cell lines: the gastric cancer cell line AZ521, the pancreatic cancer cell lines PANC-1 and Capan-1, the gall bladder cell line GBK-1, the breast cancer cell line MCF-7, and the colon cancer cell lines HT-29 and Colo205. HT-29 and Capan-1 cells producing the 600 bp RT-PCR product, were positive for mAb CA19-9. These results demonstrate that CA19-9 epitope is produced only on MUC-1/Y core protein, suggesting that CA19-9 epitope may be a specific marker for MUC-1/Y protein.     

Clinical evaluation of combined use of CEA, CA19-9 and CA50 in the serum of patients with pancreatic carcinoma

CEA, CA19-9 and CA50 are tumour-associated antigens defined by monoclonal antibodies that have been raised against adenocarcinoma cell lines, but no single antibody is specific for the detection of pancreatic malignancy. The aim of this study was to determine whether the combined use of CEA, CA19-9 and CA50 would improve diagnostic accuracy. An immunoradiometric assay was used for the detection of CEA and CA19-9 and the Delfia system for CA50. Serum was collected from 65 normal subjects, 16 with pancreatitis and 28 with pancreatic carcinoma. Of the 28 cancer patients, 24 (85%) had a CA19-9 level above 46 mu/ml, 26 (92%) had a CA50 level above 21 mu/ml and 10 (37%) had a CEA level above 7 ng/ml. Multivariant discriminant analysis on the combined antibodies showed that 96% of the malignant group, 13% of the pancreatitis group and 11% of the normal group were positive, with an overall correct classification of 91% into the three groups (multivariant discriminant analysis P less than 0.05). Thus the combined use of CEA, CA19-9 and CA50 improves diagnostic accuracy in differentiating benign from malignant disease of the pancreas.     

Serum HCG beta,CA 72-4 and CEA are independent prognostic factors in colorectal cancer

In colorectal cancer, stage is considered to be the strongest prognostic factor, but also serum tumour markers have been reported to be of prognostic value. The aim of our study was to investigate the prognostic value of serum carcinoembryonic antigen (CEA), CA 19-9, CA 242, CA 72-4 and free beta subunit of human chorionic gonadotropin (hCG beta) in colorectal cancer. Preoperative serum samples were obtained from 204 colorectal cancer patients, including 31 patients with Dukes' A, 70 with Dukes' B, 49 with Dukes' C and 54 with Dukes' D cancer. The serum levels of CEA, CA 19-9, CA 242 and CA 72-4 were measured with commercial kits with cut-off values of 5 microg/L for CEA, 37 kU/L for CA 19-9, 20 kU/L for CA 242 and 6 kU/L for CA 72-4. The serum hCG beta was quantitated by an immunofluorometric assay (IFMA) with 2 pmol/L as a cut-off value. Survival analyses were performed with Kaplan-Meier life tables, log-rank test and Cox proportional hazards model. The sensitivity was 44% for CEA, 26% for CA 19-9, 36% for CA 242, 27% for CA 72-4 and 16% for hCG beta. The overall 5-year survival was 55%, and in Dukes' A, B, C and D cancers the survival was 89%, 77%, 52% and 3%, respectively. Elevated serum values of all markers correlated with worse survival (p < 0.001). In Cox multivariate analysis, the strongest prognostic factor was Dukes' stage (p < 0.001), followed by tumour location (p = 0.002) and preoperative serum markers hCG beta (p = 0.002), CA 72-4 (p = 0.003) and CEA (p = 0.005). In conclusion, elevated CEA, CA 19-9, CA 242, CA 72-4 and hCG beta relate to poor outcome in colorectal cancer. In multivariate analysis, independent prognostic significance was observed with hCG beta, CA 72-4 and CEA.     

Tumor markers in bronchogenic carcinoma. Superiority of tissue polypeptide antigen to carcinoembryonic antigen and carbohydrate antigenic determinant 19-9

One hundred six patients with histologically proven bronchogenic carcinoma were tested for carcinoembryonic antigen (CEA), tissue polypeptide antigen (TPA), and carbohydrate antigenic determinant 19-9 (CA19-9). A total of 349 CEAs, 350 TPAs, and 317 CA19-9s were measured. In addition, sera were assayed from 57 patients with pulmonary benign diseases and their CEA, TPA, and CA19-9 levels were used as negative controls for specificity and accuracy. One hundred twenty healthy subjects provided our normal CA19-9 reference value. Sensitivity, specificity, and accuracy were obtained for CEA, TPA, and CA19-9, respectively. Significant intermarker correlations were found both at diagnosis and during follow-up, CEA and CA19-9 being the most closely related substances. The percentage of patients with elevated levels of TPA increased significantly according to tumor load. Individual values of TPA related significantly to the stage of disease. Concentrations of CEA, TPA, and CA19-9 varied significantly during the course of the illness in relation to treatment response; however, TPA showed the closest relationship to the clinical status assessments of the follow-up period. Abnormal pretreatment levels of TPA were significantly associated with a poor outcome. Biomarker combinations were clinically evaluated by calculating the mean of the percentage of the reference value for each combined marker. Using this method, any association of TPA with CEA and/or CA19-9 revealed neither a greater diagnostic accuracy nor a more reliable predictive capacity for the above clinical variables than TPA evaluated on its own. The authors believe that a single TPA assay should be added to the initial and subsequent clinical assessments of patients with bronchogenic carcinoma.     

Diagnostic value of CEA, CA 15-3, CA 19-9, CYFRA 21-1, NSE and TSA assay in pleural effusions

The aim of this study was to evaluate the individual and combined diagnostic utility of six tumor markers in patients with pleural effusion. Pleural and serum levels of carcinoembryonic antigen (CEA), carbohydrate antigen 15-3 (CA 15-3), carbohydrate antigen 19-9 (CA 19-9), cytokeratin fragment 19 (CYFRA 21-1), neuron-specific enolase (NSE) and total sialic acid (TSA) were assayed in 74 patients with pleural effusions (44 malignant and 30 benign). All tumor markers except TSA and NSE were increased in both serum and pleural fluid of patients with malignant diseases. Using the cut-off values 3 ng/ml, 14 U/ml, 5 U/ml, 8 ng/ml and 70 mg/dl for pleural fluid CEA, CA 15-3, CA 19-9, CYFRA 21-1 and TSA, respectively, the sensitivity (%) and specificity (%) of these tumor markers were as follows: CEA; 52/77, CA 15-3; 80/93, CA 19-9; 36/83, CYFRA 21-1; 91/90, TSA; 80/67, for differentiating malignant effusions from benign. When CA 15-3 and CYFRA 21-1 combined, the sensitivity and specificity were increased (100 and 83%, respectively). Classifying the malignant effusions as bronchial carcinoma and malignant pleural mesothelioma, CEA was shown to have the highest sensitivity and specificity (88 and 90%, respectively) while the combination of CEA with other tumor markers increased sensitivity but decreased specificity. According to our results, tumor markers are not suitable for the differential diagnosis of malignancy.     

Analysis of the levels of CA125, carcinoembryonic antigen, and CA19-9 in the cervical mucus for a detection of cervical adenocarcinoma

To verify whether analysis of the levels of CA125, carcinoembryonic antigen (CEA), and CA19-9 in the cervical mucus is effective for a detection of cervical adenocarcinomas or not, simultaneous measurement of these three tumor markers in the cervical mucus samples from women without gynecologic disorders, with leiomyoma, with cervical squamous cell carcinomas, and with cervical adenocarcinomas was performed. Extremely high levels of CA125 with low levels of both CEA and CA19-9 were demonstrated in the cervical mucus samples from women without gynecologic disorders and with leiomyoma. The cervical mucus samples from cervical adenocarcinomas showed low CA125 levels with extremely high CEA and/or CA19-9 levels. Therefore, analysis of the levels of these three tumor markers in the cervical mucus possibly helps in the diagnosis of cervical adenocarcinomas if CEA and/or CA19-9 show extremely high levels. When a ratio of (CEA + CA19-9)/CA125 was calculated, all women without gynecologic disorders and with leiomyoma showed a ratio less than 0.5, whereas ten of 11 cases of cervical adenocarcinomas had a ratio greater than or equal to 0.5. Only one case of microinvasive adenocarcinoma showed a ratio less than 0.5. Accordingly, the ratio greater than or equal to 0.5 strongly suggested an existence of cervical adenocarcinoma, although it included some cases of squamous cell carcinomas (four of 17 cases).     

Clinical value of tumor markers for early detection of recurrence in patients with cervical adenocarcinoma and adenosquamous carcinoma.

OBJECTIVE: The clinical value of tumor markers for early detection of recurrence was investigated in 32 patients with cervical adenocarcinoma or adenosquamous carcinoma who had recurrent tumors. METHODS: Serum levels of CA 125, CA 19-9, carcinoembryonic antigen (CEA), and squamous cell carcinoma antigen (SCC), in addition to clinical status at the time of recurrence were investigated. RESULTS: Among the 32 patients, 26 had no symptoms at the time of recurrence. In 20 patients, elevated serum levels of tumor markers were the first sign of recurrence. In 21 patients with recurrent adenocarcinoma, the positive rates were 14% (CA 125), 62% (CA 19-9), 29% (CEA), and 5% (SCC). There were 71% of cases positive for CA 19-9 and/or CEA. In 11 patients with recurrent adenosquamous carcinoma, the corresponding positive rates were 37% (CA 125), 46% (CA 19-9), 64% (CEA), and 55% (SCC), with 100% positive for CA 19-9, CEA, and/or SCC. CONCLUSIONS: The combination of CA 19-9 and CEA is probably the most promising for detection of recurrent cervical adenocarcinoma. For adenosquamous carcinoma, the additional use of SCC is recommended.