摘要:

Human γ-enolase cDNA prepared by reverse transcriptase-polymerase chain reaction was cloned into the Escherichia coli expression vector pKK223-3. The resulting plasmid, pHTK503, expressed human γ-enolase as a 46-kDa protein in SDS-PAGE, and in the cells as the active γγ form (designated as recombinant human NSE; R-NSE). R-NSE was purified from E. coli by several chromatographic elutions. Finally, 6.0 mg of R-NSE from 8.1 g cells was purified with a specific activity of 86 units/mg protein. The structural properties of R-NSE were compared with the NSE purified from human brain tissue (B-NSE). The biochemical and enzymatic characteristics were essentially the same, except for the isoelectric point (4.5 for B-NSE and 4.7 for R-NSE). In an NSE immunoassay system, R-NSE and standard NSE were almost equal in reactivity to the anti-NSE antibody. These results indicate that R-NSE can be used as standard assay mater

ISSN:1010-4283    

DOI:10.1159/000217838

出版商:S. Karger AG    

年代:1993

数据来源: Karger

摘要:

The levels of c-erbB-2 oncoprotein (ErbB-2 protein) in nipple discharge were evaluated together with those of carcinoembryonic antigen (CEA) in 9 patients with breast cancer, 2 patients with borderline lesions, 8 patients with intraductal papilloma, and 19 patients with fibrocystic change. When the tentative cutoff value was set at 40 ng/ml in the nipple discharge, elevated ErbB-2 protein levels were found in all 3 patients with palpable breast cancer and 3 of the 6 patients with nonpalpable cancer. Two of the 8 patients with intraductal papilloma had high ErbB-2 protein levels. A combination test with CEA resulted in positive detection in all cancer patients. Two patients with borderline lesions, 2 with intraductal papilloma and 2 with ñbrocystic change were positive in a combination test. In addition, elevated ErbB-2 protein levels in nipple discharge correlated well with the overexpres-sion of ErbB-2 protein in the tumor. All the patients with ErbB-2 protein levels over 100 ng/ml in their nipple discharge had comedo or solid intraductal carcinomas. Thus, measurement of ErbB-2 protein levels in nipple discharge can assist in the diagnosis of intraductal carcinoma and also in detecting tumors with a high proliferation rate and an overexpression of ErbB-2 protein: usually comedo or solid carcinomas

ISSN:1010-4283    

DOI:10.1159/000217839

出版商:S. Karger AG    

年代:1993

数据来源: Karger

摘要:

Basement membranes are frequently absent in small-cell lung cancer (SCLC). To study this phenomenon, the production of laminin by SCLC cells, both in vitro and in vivo, has been investigated and compared with the laminin production by lung carcinomas of other histotypes (non-SCLC, NSCLC). Immunoradiometric and immunoperoxidase tests, respectively, carried out on culture supernatants and cells using antilaminin rabbit antiserum, revealed that in 1 (H446) out of 8 SCLC cell lines tested and in the NSCLC line Calu3, laminin was detectable both in the culture medium and in the cytoplasm of the cells. After treatment of an SCLC-negative cell line (N592) with monensin, a molecule which inhibits protein secretion, laminin became detectable in the cytoplasm. Similar results were obtained by FACS analysis on cells permeabilized with saponin. Northern analysis indicated that the laminin B1 gene was transcribed. The level of mRNA for the B1 laminin subunit in the N592 cells was twice and 4 times higher than that found in the laminin-secreting Calu3 and H446 cell lines. The production of laminin in SCLC and NSCLC surgically resected samples using immunoperoxidase staining of cryostatic sections was also investigated. The results indicate that 85% of the NSCLC cases tested showed diffuse staining in the cytoplasm of the tumor cells and strong staining at the basement membrane level, whereas a similar staining was only found in 15% of the SCLC cases tested. The treatment of SCLC cells with differentiating agents in vitro has been shown to induce the adhesion of these cells. In fact, n-butyric acid induced the disaggrega-tion of floating-clump cells and single-cell adhesion in the N592 line, whereas after treatment with retinoids the clumps of the N592 cells were still present, but 30% of these were found to adhere to the plate. However, no increase in the laminin production was found in the cytoplasmic or culture medium under these conditions.

ISSN:1010-4283    

DOI:10.1159/000217840

出版商:S. Karger AG    

年代:1993

数据来源: Karger

摘要:

In vitro studies were performed to examine the antitumor effect of protein-doxorubicin (DXR) conjugate on the growth of the multidrug-resistant human chronic myelogenous leukemia cell line, K562/DXR. The 50% inhibitory concentration (IC50) for DXR in the K562/DXR cell line was 20 nM (in the K562 parental cell line, IC50 was 3.2 nM). Treatment of both types of cells with various concentrations of DXR or conjugates at equivalent concentrations of DXR was carried out. One type of the conjugates used was human serum albumin (HSA)-DXR conjugate and human transferrin (Tf)-DXR conjugate via a glutaraldehyde bridge (HSA-ga-DXR, Tf-ga-DXR, respectively) and another type used was HSA-DXR conjugate with a dextran bridge (HSA-dex-DXR). All of these conjugates showed potent dose-dependent inhibition of cell growth against the K562/ DXR cells as compared with the cells treated with DXR or other controls. IC50 for HSA-ga-DXR, Tf-ga-DXR and HSA-dex-DXR conjugates in the K562/DXR cell line was 2.4, 3.6 and 1.0 (equivalent DXR) nM, respectively, which were approximately similar to the value of the K562 treated with DXR. Through the treatment of K562/DXR cells with HSA-DXR conjugate, the intracellular drug concentration increased as a function of time up to 24 h compared with the cells treated with DXR. Intracellular DXR effluxed rapidly from K562/DXR cells, but HSA-ga-DXR as well as HSA-dex-DXR conjugates remained in the cells at a relatively high concentration for a long time. These results indicate that it may be possible to overcome multidrug resistance by chemically modifying DXR, such as by conjugation of the drug with proteins.

ISSN:1010-4283    

DOI:10.1159/000217841

出版商:S. Karger AG    

年代:1993

数据来源: Karger

摘要:

To determine whether c-kit and kit ligand (KL) mRNAs could be expressed in human epithelial tumors, reverse transcrip-tase-polymerase chain reaction and Northern blot analysis were performed. KL mRNA was shown to be expressed in a variety of epithelial tissues and cell lines. The expression of c-kit mRNA was then examined in hepatocellular and colon carcinoma cell lines. While hepatocellular carcinoma cell lines did not express c-kit mRNA as far as we could ascertain, 2 of 5 colon carcinoma cell lines showed the expression of both c-kit and KL mRNAs. Furthermore, the expression of c-kit in these cells was demonstrated at the protein level by flow cytometry. These data suggest that c-kit and KL may play an important role as an autocrine loop in the proliferation of some colon carcinoma cells.

ISSN:1010-4283    

DOI:10.1159/000217842

出版商:S. Karger AG    

年代:1993

数据来源: Karger

摘要:

Preoperative serum soluble interleukin-2 receptor (sIL·2R) and CA 125 levels were measured in 183 patients with ovarian masses undergoing laparotomy. Serum sIL·2R levels were higher in the 54 patients with epithelial ovarian cancer than in the 129 patients with benign ovarian diseases (p < 0.0001). Elevated serum levels of sIL-2R (≥71 U/ml) and CA 125 (≥ 83 U/ml) were found in 79.6 and 77.8% of patients with epithelial ovarian cancer, respectively. Serum sIL-2R and CA 125 positivity rates correlated with the FIGO stage (p = 0.0033 and p = 0.0001, respectively). Raised serum levels of sIL-2R and CA 125 were detected in 11.6 and 7.0% of patients with benign ovarian diseases, respectively. The combination sIL-2R or CA 125 had a sensitivity of 88.9%, and the association sIL·2R and CA 125 had a specificity of 98.4% for epithelial ovarian cancer. As for the 16 patients with this malignancy who were serially monitored during and after chemotherapy, changes in sIL-2R and CA 125 levels correlated with the clinical course of disease in 62.3 and 94.3% of 53 instances, respectively. In conclusion, the serum sIL·2R assay could represent a useful adjunctive tool for the differential diagnosis of ovarian masses, while it seems to be of limited benefit for monitoring the response to chemotherapy and follow-up of patients with epithelial ovarian

ISSN:1010-4283    

DOI:10.1159/000217843

出版商:S. Karger AG    

年代:1993

数据来源: Karger

摘要:

CA 125 has been established as an important tumor marker for monitoring patients diagnosed with nonmucinous ovarian cystadenocarcinoma although it has also been shown to be expressed by other carcinomas, normal epithelial tissues, and fetal tissues. Current evidence implicates a role for CA 125 during early fetal development. The human epithelial amnion WISH cell line is a known secretor of CA 125. WISH cells have been investigated as a model system to characterize the structure of cell-associated and secreted CA 125 of fetal origin. CA 125 secretion was maximal in confluent monolayers of WISH cells where it averaged 2,081 units/ml/24 h. Secretion ranges from 600 to 900 units/106 cells/24 h. [35S]-Methionine-la-belled CA 125 can be detected by 4 h and reached maximal levels of radioactive incorporation in tissue culture medium by 12 h when analyzed by immunoprecipitation with the Ml 1 anti-CA 125 monoclonal antibody and SDS-PAGE, followed by autoradiography. Both cycloheximide and actinomycin D inhibited CA 125 synthesis. CA 125 was demonstrated to incorporate [3H]-galactose but the level of radioactive incorporation was greatly reduced when WISH cells were incubated in the presence of phenyl N-acetyl-α-D-galactosaminide (an inhibitor of O-linked glycosylation) or monensin (an inhibitor of intracellular protein transport within the Golgi complex). Treatment of WISH cells with tunicamycin (an inhibitior of N-linked glycosylation) only slightly decreased label incorporation

ISSN:1010-4283    

DOI:10.1159/000217844

出版商:S. Karger AG    

年代:1993

数据来源: Karger

摘要:

Serum α-fetoprotein (AFP) from cord blood and from patients with hepatitis, cirrhosis, hepatocellular carcinoma, gastrointestinal tumors and yolk sac tumor was analyzed by extended agarose gel electrophoresis coupled with our sensitive detection method of antibody-affinity blotting. AFP was separated into AFP-A, AFP-B and AFP-C from the anode to the cathode, corresponding to disialo-, monosialo- and asialo-AFP, respectively, as revealed by the results of neuraminidase digestion of serum AFP. AFP-Af, which migrated ahead of AFP-A as band or leading smear and varied widely in intensity, was eliminated in calculating the proportions of other band intensities. Disialo-AFP was the major component and mono-sialo-AFP the minor one in benign conditions, the latter being 4.2 ± 6.0% in chronic hepatitis and 9.0 ± 8.9% in cirrhosis. Monosialo-AFP in hepatocellular carcinoma was increased significantly, the proportion being 29.9 ± 11.2%. AFP in other malignancies was further characterized by the appearance of asialo-

ISSN:1010-4283    

DOI:10.1159/000217845

出版商:S. Karger AG    

年代:1993

数据来源: Karger