bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of the bcl‐2 gene family in normal and malignant breast tissue: Low bax‐α expression in tumor cells correlates with resistance towards apoptosis
Authors: Bargou Railf C. , Daniel Peter T. , Mapara Markus Y. , Bommert Kurt , Wagener Christian , Kallinich Bianca , Royer Hans D. , Dörken Bcrnd
Published: February 2007

We have studied the expression of the apoptosis‐regulating genes bcl‐2, bcl‐x, bax and APO‐1/fas (CD95) in human breast cancer. The expression pattern of these genes in human breast‐cancer tissues and breast‐cancer‐derived cell lines was compared to that seen in normal breast epithelium and breast epithelial cell lines. No difference with regard to bcl‐2 and bcl‐xL expression was observed between normal breast epithelium and tumor tissue or breast cancer and non‐malignant epithelial cell lines. In contrast, bax‐α, a splice variant of bax, which promotes apoptosis, is expressed in high amounts in normal cell lines and breast tissue, whereas only weak or no expression could be detected in cancer‐cell lines and malignant tissue. In contrast to malignant cell lines, which express low levels of bax‐α, non‐malignant epithelial cell lines displaying high amounts of bax‐α were highly sensitive to induction of programmed cell death by both serum starvation and APO‐1/fas (CD95) triggering. We therefore propose that dysregulation of apoptosis contributes to the pathogenesis of breast cancer, at least in part, due to an imbalance between anti‐apoptosis genes (such as bcl‐2/bcl‐x) and apoptosis‐promoting genes (bax).
High cell kinetics is associated with amplification of the int‐2, bcl‐1, myc and erbB‐2 proto‐oncogenes and loss of heterozygosity at the DF3 locus in primary breast cancers
Authors: Contegiacomo Alma , Pizzi Claudia , De Marchis Laura , Alimandi Maurizio , Delrio Paolo , Palma Ester Di , Petrella Giuseppe , Ottini Laura , French Deborah , Frati Luigi , Bianco Angelo R. , Mariani‐Costantini Renato
Published: February 2007

Cell kinetics is a predictive parameter of breast‐cancer aggressiveness, and mutations occurring in mammary tumorigenesis may favor uncontrolled cell proliferation. In this study, cell kinetics, clinico‐pathological characteristics and genetic alterations at the int‐2, bcl‐1, c‐myc, c‐erbB‐2, and DF3 loci were analyzed and correlated in 54 primary breast carcinomas. The occurrence of mutations at more than one locus was also studied. Tumor‐proliferative activity was evaluated by determination of the thymidine labeling index (TLI). Amplification (AMP) of int‐2 was observed in 11.2%, of bcl‐1 in 9.4%, of c‐myc in 5.7% and of c‐erbB‐2 in 8.6% of the carcinomas. Loss of heterozygosity (LOH) at the DF3 locus was detected in 13.9% of the tumors. Genetic alterations demonstrated a significant association with patient's age and high TLI values. AMP and LOH + AMP did not appear to be statistically related to histotype, histological grade, tumor size or lymph‐node status. Alone, allele loss at the DF‐3 locus was not significantly associated with any of the clinico‐pathological characteristics studied. Alterations at more than one locus, including int‐2/bcl‐1, int‐2/ c‐myc, int‐2/bcl‐1/c‐erbB‐2, and c‐myc/DF3, were detected in 11.1% of the tumors. Multiple mutations were found only in less differentiated tumors, which included the 2 cases from the youngest patients of the series. © 1995 Wiley‐Liss, Inc.
Cancer incidence and mortality among teenagers in vaud, switzerland, 1974–1992
Authors: Levi Fabio , La Vecchia Carlo , Randimbison Lalao , Te Van‐Cong
Published: February 2007

Data from the Vaud Cancer Registry, Switzerland, were used to analyse incidence and mortality from cancer in teenagers (aged 10 to 19 years) over the period 1974–1992. A total of 113 males and 87 females were registered. Of these, 23% were lymphomas, 16% leukaemias, about 15% central nervous system neoplasms, 10% germ cell tumours and bone neoplasms and 8% soft tissue sarcomas. The overall incidence rate (age‐adjusted, world standard population) for all cancers combined was 167 per million boys and 128 per million girls. In both sexes, there was some indication of rising trends over time, to reach 196 per million males and 141 per million females in 1986–1992. The only types of cancer showing consistent upward trends in both sexes were lymphomas. A total of 53 cancer deaths were certified, due to leukaemias in about 40% of cases in both sexes and to lymphomas and brain tumours in 15%–20%. The overall mortality rate at age 10–19 years (age‐standardised, world standard) was 47 per million boys and 31 per million girls, and no trend in mortality was observed over time. The 2 main findings of our analysis are (i) the absence of major trends in cancer incidence in adolescents, with the sole exception of a possible increasing incidence of lymphomas, and (ii) the lack of appreciable trends in mortality, in view of the declines in cancer mortality observed in children and young adults over the same calendar period. © 1995 Wiley‐Liss, Inc.
Characterization of mature and immature Rad LV‐Induced thymic T‐cell lines for tumorigenesis and mhc‐class‐I gene expression
Authors: Bashi Orna , Ehrlich Rachel
Published: February 2007

Class‐I‐MHC molecules are divided into class‐la molecules, which play major roles in recognition of virus‐infected cells, graft rejection and immune responses against tumors, and class‐lb molecules, which are less polymorphic and may be responsible for presenting unique classes of peptides. Our report characterizes RadLV‐induced thymic T‐cell lines that differ both in their tumorigenic potential and in the level of protein for class‐la and TL genes. The PDI.I cell line is CD4−CD8+ and expresses relatively high levels of class‐l as compared with the CD4+CD8+ PDI.2 cell line. These class‐I‐expression levels correlate with thymocytes and splenic T cells of the same phenotype, except that normal cells fail to express TL3b. Interferon‐treated PDI.2 cells demonstrate significantly lower levels of class‐l expression than do interferon‐treated PDI.I cells, and were shown to contain large amounts of degraded class‐l mRNA, at least some of which was TL in origin. These RNA products were not detected in PDI.I cells, suggesting the existence of a mechanism controlling cell‐specific and gene‐specific mRNA stability. Such RadLV‐induced cell lines provide a means for obtaining stage‐specific T cells, which can be used for studying the regulation of class‐l gene expression during T‐cell differentiation, as well as factors that differentially regulate class‐la and class‐lb expression and are potentially useful for studying T‐cell differentiation in general. © 1995 Wiley‐Liss, Inc.
Glycosylation of a novel member of the immunoglobulin gene superfamily expressed in rat carcinoma cell lines
Authors: Chadéneau Corinne , Le Moullacguy Cornu Béatrice , Meflah Khaled , Denis Marc G.
Published: February 2007

MAb E4 recognizes a 66‐kDa glycoprotein, pE4, which is a member of the immunoglobulin gene superfamily. This protein is expressed at the cell surface in rat colon and mammary carcinomas, but only in trace amounts in normal adult rat tissues. Since expression of aberrant carbohydrate structures is often associated with malignant transformation, glycosylation of pE4 was analyzed. Reactivity of lectins with pE4 suggested the absence of N‐acetylneuraminic acid, terminal galactose and O‐linked glycan, and the presence of N‐linked glycans. Tunica‐mycin treatment reduced the binding of MAb E4 to cancer cells suggesting that the E4 epitope is at least partially glycosylated. Digestions with neuraminidases, O‐glycosidase and peptide‐N‐glycosidase F confirmed these results. Pronase treatment abolished the binding of MAb E4, indicating that E4 epitope involves not only a carbohydrate determinant but also a peptide moiety. Mild periodate oxidation abolished the binding of MAb E4, indicating that non‐reducing terminus carbohydrates are part of the E4 epitope. Neutral sugar analysis revealed the absence of galactose and the presence of fucose. Since fucose is sensitive to periodate oxidation, this sugar could be the carbohydrate part of the determinant recognized by MAb E4. Reactivity of lectins specific for fucose indicated the presence of ø(1‐6)‐fucose on pE4. © 1995 Wiley‐Liss, Inc.
Direct transforming activity of TGF‐β on rat fibroblasts
Authors: Vossbeck Heinke , Höfler Brigitte Strahmpetra , Bauer Georg
Published: February 2007

Treatment of NRK 536 fibroblasts with EGF and transforming growth factor type beta (TGF‐β) is known to lead to the reversible induction of the transformed phenotype in a large percentage of cells. Maintenance of the transformed state is dependent on the continuous presence of the cytokines. Here we show that treatment of these cells with TGF‐β alone leads to the formation of a small percentage of colonies, the majority of which stably retain the transformed phenotype after removal of the cytokine. Colony induction is dependent on the concentration of TGF‐β originally present. Stably transformed cells grow in soft agar without further addition of exogenous TGF‐β, and exhibit criss‐cross morphology in monolayer culture and a diffuse actin pattern. Transformation of NRK 536 cells can be demonstrated only for individualized cells treated with TGF‐p in soft agar, as treatment in monolayer leads to the induction of apoptosis in newly transformed cells by neighboring normal cells. Transformation of NRK 536 fibroblasts by TGF‐β is not due to the induction of point mutations by TGF‐β. © 1995 Wiley‐Liss, Inc.
The human immunodeficiency virus(HIV) inhibitor 9‐(2‐phosphonylmethoxyethyl)adenine (pmea) is a strong inducer of differentiation of several tumor cell lines
Authors: Balzarini J. , Verstuyf A. , Hatse S. , Goebels J. , Sobis H. , Vandeputte M. , De Clercq E.
Published: February 2007

9‐(2‐phosphonylmethoxyethyl)adenine (PMEA) is the prototype compound of a series of acyclic nucleoside phosphonate derivatives endowed with potent and selective anti‐retroviral activity in vitro and in vivo. We have now found that PMEA is also a potent inducer of differentiation of a number of tumor cells, including human erythroleukemia K562 cells, rat choriocarcinoma RCHO cells and human acute promyelocytic leukemic HL‐60 cells. At 10 μM PMEA, rat RCHO cell cultures could be almost fully differentiated, and at 50 μM PMEA, approximately 50% of the K562 cells could be triggered to produce hemoglobin. The differentiating activity of butyric acid was at least partially additive to that of PMEA when both drugs were combined in K562 cell cultures. PMEA needs to be present for at least 2 or 3 days in the K562 cell cultures to achieve optimal differentiating activity. This suggests that either a PMEA metabolite and/or its anti‐metabolic effects may be responsible for differentiation of the tumor cells. © 1995 Wiley‐Liss, Inc.
Muscle wasting associated with cancer cachexia is linked to an important activation of the atp‐dependent ubiquitin‐mediated proteolysis
Authors: Llovera Marta , Garcia‐Martinez Cèlia , Agell Neus , Lopez‐Soriano Francisco J. , Argiles Josep M.
Published: February 2007

Rats bearing the Yoshida AH‐130 ascites hepatoma for 7 days showed an important decrease in muscle mass—over 30% in gastrocnemius and extensor digitorum longus (EDL)—in relation to non‐tumour‐bearing controls, which is associated with an increased proteolytic rate‐in in vitro incubation. In order to identify the precise biochemical process which was involved, we measured different proteolytic systems in incubated EDL muscles. The capacity for intralysosomal proteolysis, as measured by sensitivity to methylamine, was not increased in tumour‐bearing rats, suggesting that the mechanism involved in the increased proteolytic rate was extralysosomal. Incubations using the Ca2+ ionophore A23187 revealed no change in the activity of calcium‐dependent proteases as a consequence of tumour growth. Finally, muscle incubation in an ATP‐depleted medium allowed us to conclude that energy‐dependent proteases were involved in the activation of muscle proteolysis in tumour‐bearing rats. In particular, the ubiquitin‐dependent proteolytic system is involved, since there is an important increase in ubiquitin conjugates in the skeletal muscle of tumour‐bearing rats. It may thus be suggested that extralysosomal ATP‐and ubiquitin‐dependent proteases underlie the biochemical mechanism of muscle wastage associated with cancer cachexia. © 1995 Wiley‐Liss, Inc.
Anti‐gastrin antibodies raised by gastrimmune inhibit growth of the human colorectal tumour AP5
Authors: Watson Susan A. , Michaeli Dov , Grimes Steven , Morris Teresa M. , Crosbee David , Wilkinson Mark , Robinson Graham , Robertson John F. R. , Steele Robert J. C. , Hardcastle Jack D.
Published: February 2007

The neutralising ability of rabbit anti‐gastrin‐17 (G17) anti‐serum raised by Gastrimmune, an immunogen constructed of the N‐terminal portion of human G17 conjugated to diptheria toxoid (DT), was evaluated. The anti‐serum (denoted anti‐G17: DT) was shown to displace 125[I] G17 from the gastrin receptors on AR42 cells. The therapeutic effect of the rabbit anti‐G17:DT anti‐serum was evaluated on a freshly derived human colorectal cancer cell line, AP5, which was shown to express both gastrin receptors and gastrin immunoreactivity as assessed by immunocytochemistry. Rabbit anti‐G17:DT anti‐serum was shown to block basal in vitro growth of AP5 cells when used at an antigen binding capacity of 3.75 × 10−9 M. The same dilution of anti‐serum completely reversed growth stimulated by human G17 at concentrations of 1 × 10−8 and 1 × 10−9 M but did not inhibit growth at 1 × 10−8 M G17. When AP5 was grown as a xenograft in nude mice, the sensitivity to the proliferative effect of human G17 was maintained. In addition, the basal growth of AP5 xenografts was significantly reduced by i.v. infusion of rabbit anti‐G17:DT anti‐serum when compared to treatment with rabbit anti:DT control anti‐serum. Thus anti‐G17:DT antibodies raised by Gastrimmune may be of clinical value in gastrin‐sensitive tumours. © 1995 Wiley‐Liss, Inc.
O6‐methylguanine‐DNA methyltransferase activity in breast and brain tumors
Authors: Preuss Ilka , Eberhagen Ingrid , Haas Simone , Eibl Robert H. , Kaufmann Manfred , Minckwitz Gunther Von , Kaina Bernd
Published: February 2007

The DNA repair protein O6‐methylguanine‐DNA methyltransferase (MGMT) is a main determinant of resistance of tumor cells to the cytostatic activity of chemotherapeutic alkylating agents (methylating and chloroethylating nitrosoureas) and is effective in protecting normal cells against genotoxic and carcinogenic effects resulting from DNA alkylation. Therefore, the level of expression of MGMT is significant for the response of both the tumor and the non‐target tissue following application of nitrosoureas in tumor therapy. To determine the expression of MGMT in tumor tissue, we have assayed MGMT activity in 68 breast carcinomas and 38 brain tumors. There was a wide variation of MGMT expression in breast carcinomas ranging from below the level of detection up to 863 fmol/mg protein. About 4% of breast tumors did not display detectable MGMT, 15% had activity lower than 100 fmol/mg protein, and 26% expressed more than 500 fmol/mg. The mean level of expression was 321 fmol/mg. In brain tumors (astrocytoma WHO grade I, II, and III, and glioblastoma WHO grade IV) the MGMT activity was generally lower than in breast tumors, ranging from below the level of detection up to 238 fmol/mg. The mean level of expression was 55 fmol/mg. Five percent of the brain tumors had no detectable MGMT activity. The MGMT repair activity correlated well with the amount of MGMT protein present in tumor samples, as shown by Western‐blot analysis, indicating that loss of MGMT repair activity is due to inability of these tumor cells to synthesize the protein. © 1995 Wiley‐Liss, Inc.
Epstein‐barr virus (EBV) in malaysian upper‐aerodigestive‐tract lymphoma: Incidence and sub‐type
Authors: Peh Suat C. , Sandvej Kristian , Pallesen Gorm
Published: February 2007

Epstein‐Barr virus (EBV) type B, a less potent transformer of B lymphocytes than type A, has rarely been detected in EBV‐associated neoplasms except in AIDS‐related lymphomas, in which about 50% of the cases contained this sub‐type. In this study we analyzed the association of EBV and the distribution of virus sub‐types in Asian non‐Hodgkin's lymphoma (NHL) of the upper aerodigestive tract. We studied archival material of 29 NHL cases from Malaysia. B‐ and T‐cell associated antigens were demonstrated by immunohistochemistry, and EBV early RNA EBER‐1 was demonstrated using the RNA in situ hybridization technique. EBV was detected in the majority of tumour cells in 11/13 T‐NHL but in only 1/16 B‐NHL. EBV was sub‐typed by single‐step polymerase chain reaction of the EBNA‐2 gene. This was successful in 9/10 cases of EBER‐I‐positive tumours and all contained type‐A virus only. Our results showed a preponderance of T‐cell lymphoma of the upper aerodigestive tract in the ethnic Chinese group of Malaysian patients, and EBV was strongly associated with T‐NHL but not with B‐NHL. Our results suggest that type‐A EBV is the prevalent sub‐type in Asian NHL of the upper aerodigestive tract, similarly to findings in Asian nasopharyngeal carcinoma. © 1995 Wiley‐Liss, Inc.
Incidence and prognostic significance of vascular and neural invasion in squamous cell carcinomas of the esophagus
Authors: Sarbia Mario , Porschen Rainer , Borchard Franz , Horstmann Olaf , Willers Rainhart , Gabbert Helmut E.
Published: February 2007

The prognostic influence of blood‐vessel invasion (BVI), lymphatic‐vessel invasion (LVI) and neural invasion (NI) was evaluated retrospectively in a series of 161 patients with squamous cell carcinoma (SCC) of the esophagus who underwent esophageal resection. Evidence of BVI, LVI and NI was found in 32.9%, 48.5% and 26.1%, respectively. Incidence of BVI, LVI and NI was significantly higher in high pT categories (pT3 and pT4) than in low pT categories (pTI and pT2) and in patients with distant metastases than in patients without distant metastases. Incidence of LVI and NI in lymph‐node‐positive patients was significantly higher than in lymph‐node‐negative patients. The 5‐year survival rate was significantly lower in patients with BVI or LVI than in patients without BVI or LVI. Patients with evidence of NI showed no significant differences in 5‐year survival from patients without evidence of NI. By stepwise multivariate Cox regression analysis, BVI and LVI were shown to be independent prognostic factors. A search for vascular invasion may therefore provide additional prognostic precision in SCC of the esophagus. © 1995 Wiley‐Liss, Inc.
Resistance‐associated factors in human small‐cell lung‐carcinoma GLC4 sub‐lines with increasing adriamycin resistance
Authors: Versantvoort C. H. M. , Withoff S. , Broxterman H. J. , Kuiper C. M. , Scheper R. J. , Mulder N. H. , De Vries E. G. E.
Published: February 2007

Previous studies have shown that the in vitro‐selected adriamycin‐resistant human small‐cell lung‐carcinoma cell line GLC4‐ADR150 displays multidrug resistance as the result of 3‐fold decreased DNA‐topoisomerase II (topo II) activity and a 6‐fold reduction in adriamycin accumulation. Not the MDRI gene, but the MRP gene, was over‐expressed in this cell line. The aim of our study was to establish which of these drug‐resistance‐associated factors are already involved in drug resistance occurring at early steps of selection with adriamycin. To address this question, changes in expression of topo IIα/topo IIβ, MRP and drug accumulation were measured along with adriamycin resistance (from 2‐ to 10‐ to 150‐fold) and in a partial revertant cell line (10‐fold resistant). Topo IIα and IIβ mRNA and protein levels were decreased in the resistant sub‐lines, except in the 10‐fold‐resistant cell line. Cellular daunorubicin accumulation was decreased 1.2‐ to 5‐fold with increasing resistance. MRP mRNA was over‐expressed in all resistant sub‐lines, with a marked increase in the 10‐fold‐resistant cells (level of expression as high as in the GLC4‐ADR150 cells). Expression of an ATP‐binding 190‐kDa membrane protein and Western‐blot analysis with anti‐MRP anti‐serum ASPKE, was in accordance with the expression of MRP mRNA in all cell lines. Expression of MRP mRNA and protein, however, was not proportional with the decrease in drug accumulation in all resistant sub‐lines. This study also shows that drug accumulation, topo II and MRP expression were all changed at the earliest stage of resistance development of GLC4 cells upon adriamycin selection. © 1995 Wiley‐Liss, Inc.
Glycosylation of β‐1 integrins in B16‐F10 mouse melanoma cells as determinant of differential binding and acquisition of biological activity
Authors: Veiga Silvio S. , Chammas Roger , Cella Nathalie , Brentani Ricardo R.
Published: February 2007

Studying B16‐F10 cells we could identify β‐1 integrins as laminin, fibronectin and collagen receptors. Gradient ionic strength elution analysis of affinity chromatography showed differential interactions between laminin‐binding β‐1 integrins (two β‐1 polypeptides of 105 and 120 kDa) and fibronectin and collagen‐binding β‐1 integrins (elution of one major β‐1 polypeptide of 120 kDa) and their respective ligands. To evaluate this diversity we submitted B16‐F10 extracts to IEF and SDS‐PAGE and found that one β‐1 integrin formed acidic and larger isoforms, while another formed basic and smaller isoforms. To study this difference we also submitted material eluted from WGA‐Sepharose columns to IEF but now only the acidic β‐1 isoform was found. Extracts of B16‐F10 treated with neuraminidase showed only the basic β‐1 isoform, suggesting that terminal sialic acid residues may be responsible for this acidic pattern, an interpretation supported by the fact that MAA (Maackia ammurensis agglutinin) reacts only with the acidic isoform. Differential glycosylation of β‐1 integrin isoforms in B16‐F10 was also demonstrated since the smaller lamininbinding β‐1 integrin isoform reacted only with GNA (Galanthus nivalis agglutinin), whereas the mature larger form reacted with DSA (Datura stramonium agglutinin) and MAA; thus this heterogeneity of β‐1 chains is essentially due to variable glycosylation. Autoradiography and immunoblotting analysis of material separated by 2‐dimensional electrophoresis show that only the processed forms of β‐1 integrins are expressed at the cell surface. © 1995 Wiley Liss, Inc.
Tenascin‐C in serum: A questionable tumor marker
Authors: Schenk Susanne , Muser Jürgen , Vollmer Günter , Chiquet‐Ehrismann Ruth
Published: February 2007

In normal adult tissue tenascin‐C (TN‐C) is usually expressed at low levels. However, it is strongly induced in many tumors as well as in other pathological conditions often associated with inflammation. To evaluate the diagnostic significance of TN‐C, we established a sensitive sandwich ELISA to determine TN‐C levels in serum. Furthermore, we investigated the distribution of TN‐C variants in serum and found the large TN‐C isoforms to be predominant. We measured TN‐C in sera from 15 healthy persons, 75 tumor patients and 84 patients selected due to their elevated levels of the acute‐phase protein C‐reactive protein (CRP), which is a very specific marker for infection and inflammation. It was found that sera from cancer patients can have elevated TN‐C levels; however, the increase was more pronounced in persons with high levels of CRP. There appeared to be a correlation of TN‐C levels with the levels of CRP. In view of these facts, the diagnostic value of TN‐C levels in serum as a potential tumor marker seems to be questionable, since our data show that TN‐C levels can be elevated as a consequence of infection and inflammation. © 1995 Wiley‐Liss, Inc.
Human papillomaviruses in 91 oral cancers from indian betel quid chewers—high prevalence and multiplicity of infections
Authors: Balaram Prabha , Nalinakumar K. R. , Abraham Elizabeth , Balan Anita , Hareendran N. K. , Bernard Hans‐Ulrich , Chan Shin‐Yen
Published: February 2007

India has one of the world's highest incidences of oral cancer. The habit of chewing betel quid is widespread and is suspected to play a role in the etiology of this disease. Studies in many other countries have also pointed to a role for human papilloma‐viruses (HPVs) in the etiology of some oral cancers. In this study we analyzed biopsies from 91 Indian oral cancer patients, most of whom were betel quid chewers, by PCR amplification and direct DNA sequencing. HPV DNA was detected in 74% of these lesions, of which 41% had multiple HPV infections. Among the lesions from different oral sites, lesions of the tongue had the highest rate (9 of 11) of HPV infection. These HPV prevalences are among the highest ever reported in oral cancers. As to individual HPV types, prevalences of HPV‐6, HPV‐II, HPV‐16 and HPV‐18 were 13%, 20%, 42% and 47%, respectively. No additional known or novel HPV types were detected. To understand the unexpectedly high prevalences of the “low‐risk” types HPV‐6 and HPV‐11, we compared the subtypes and variants that were found in oral cancers against those from benign genital warts from the same patient population but found no differences. The high prevalence of HPV in the oral cancers of these Indian patients suggests that viral infection is an important etiological component, with betel quid probably causing additional mutagenic steps in the carcinogenic process. © 1995 Wiley‐Liss, Inc.
αv‐Integrins in human melanoma: Gain of αvβ3 and loss OF αvβ5 are related to tumor progression in situ but not to metastatic capacity of cell lines in nude mice
Authors: Danen Erik H. J. , Jansen Kees F. J. , Van Kraats Annemieke A. , Cornelissen Ine M. H. A. , Ruiter Dirk J. , Van Muijen Goos N. P.
Published: February 2007

We investigated the expression of αv‐integrins in different stages of human cutaneous melanocytic tumor progression. We observed that αvβ5 was the αv‐integrin expressed in all common nevocellular nevi, in 78% of dysplastic nevi, in 63% of early primary melanomas, in 43% of advanced primary melanomas, and in 33% of melanoma metastases. Hence, loss of αvβ5 expression was related to melanocytic tumor progression. In line with earlier reports, αvβ3 was exclusively detected in advanced primary melanomas and metastases (24% and 50% respectively). Staining with anti‐αv monoclonal antibodies (MAbs) in lesions where both αvβ3 and αvβ5 were absent showed that alternative αv‐integrins were expressed in advanced primary melanomas and metastases. By FACS analysis, we determined expression of αvβ5 and αvβ3 in 4 human melanoma cell lines with different metastatic capacities after s.c. inoculation into nude mice. One of the non‐metastatic and both highly metastatic cell lines expressed αvβ5 at their surface. Surprisingly, αvβ3 was detected exclusively in the non‐metastatic cell lines. Absence of αvβ3 in the highly metastatic cell lines was confirmed by lack of immunoprecipitation from 35S‐methionine‐labeled cells and by absence of immunohistochemical staining on primary and metastatic xenograft lesions. Our findings indicate that αvβ5 expression is often lost in advanced stages of melanocytic tumor progression in situ, while αvβ3 is acquired, but that a decrease in αvβ5 and an increase in αvβ3 expression are not necessarily related to the metastatic behavior of human melanoma cells in nude mice. © 1995 Wiley‐Liss, Inc.
Bispecific monoclonal antibody anti‐CD3 X anti‐tenascin: An immunotherapeutic agent for human glioma
Authors: Bonino Laura Davico , De Monte Lucia B. , Spagnoli Giulio C. , Vola Roberto , Mariani Massimo , Barone Domenico , Moro Ana M. , Riva Pietro , Nicotra Maria R. , Natali Pier Giorgio , Malavasi Fabio
Published: February 2007

Besides surgery, the therapeutic possibilities for the treatment of human gliomas include adoptive cellular immuno‐therapy, radioimmunotherapy, immunotherapy mediated by chemoimmunoconjugates and, more recently, bispecific monoclonal antibodies (biMAbs). Anti‐CD3 x anti‐tenascin (TN) is the first reagent of a number of biMAbs under investigation for prospective use in vivo to maximize the cell‐mediated cytolytic potential of glioma patients. This biMAb originated from the fusion of 2 parental hybridomas, made resistant by retrovirus‐mediated infection to the different metabolic drugs, geneticin and methotrexate, respectively. The resulting hybrid hybridomas were selected on the basis of the double specificity for CD3 and TN, cloned several times and grown under continuous metabolic pressure. The different families of recombinant antibodies were then purified by high‐pressure liquid chromatography on hydroxylapatite columns. Immunohistochemical studies on tumor specimens of different origin and histotype have shown that the selected biMAb presented a distribution pattern similar to that of the parental anti‐TN MAb, maintaining the same staining homogeneity and intensity. Moreover, the mito‐genic activity of anti‐CD3 x anti‐TN biMAb on peripheral blood mononuclear cells was similar to that featured by the parental anti‐CD3 MAb. Furthermore, the hybrid molecule induced TNF‐α gene expression in activated PBMC. Finally, the anti‐CD3 x anti‐TN featured the desired killer targeting ability, being able to induce a significantly increased cytotoxic activity against TN† tumor cells. © 1995 Wiley‐Liss, Inc.
Selective inhibition of spontaneous pulmonary metastasis of lewis lung carcinoma by 5′‐deoxy‐5‐fluorouridine
Authors: Ishikawa Tohru , Ura Masako , Yamamoto Taeko , Tanaka Yutaka , Ishitsuka Hideo
Published: February 2007

5′‐deoxy‐5‐fluorouridine (5′‐FUdR) is a cytostatic that is biotransformed to 5‐fluorouracil (5‐FUra) by pyrimidine nucleo‐side phosphorylase (PyNPase), the expression of which is up‐regulated by tumor necrosis factor α (TNFα), interleukin‐1 α (IL‐1α) and interferon γ (IFNγ). In Lewis lung carcinoma (LLC) cell cultures, these inflammatory cytokines up‐regulated the expression of type‐IV collagenase, a metastatic factor, as well as PyNPase and consequently enhanced the antiproliferative activity of 5′‐FUdR. However, the activity of 5‐FUra was not enhanced. It appears that 5′‐FUdR selectively kills highly metastatic cells which are exposed to these intrinsic cytokines in tumor tissues, because of their high PyNPase activity. In fact, 5′‐FUdR inhibited the spontaneous metastasis of LLC from the s.c. inoculation site to the lung. When 5′‐FUdR was given during the process of metastasis, it greatly reduced the number of tumor nodules in the lung even at doses 46 times lower than those inhibiting the primary tumor growth. In addition, 5′‐FUdR, but not 5‐FUra, lowered type‐IV collagenase levels in the tumors at the low dose showing only anti‐metastatic activity. On the other hand, 5‐FUra showed anti‐metastatic activity at doses similar to or only several times lower than those inhibiting the primary tumor growth. © 1995 Wiley‐Liss, Inc.
Expression of protectin (CD59) in human melanoma and its functional role in cell‐ and complement‐mediated cytotoxicity
Authors: Brasoveanu Lorelei I. , Altomonte Maresa , Gloghini Annunziata , Fonsatti Ester , Coral Sandra , Gasparollo Aldo , Montagner Roberto , Cattarossi Ilaria , Simonelli Cecilia , Cattelan Alessandro , Attadia Vincenza , Carbone Antonino , Malo Michele
Published: February 2007

Immunohistochemical and/or indirect immunofluorescence analysis with monoclonal antibody (MAb) H19 demonstrated the expression of protectin (CD59) in 54 surgically removed metastatic melanoma lesions and on 8 out of 12 melanoma cell lines. CD59 expression had a low degree of intra‐ and intertumor heterogeneity. SDS‐PAGE analysis showed that the molecular weight of CD59 expressed on melanoma cells is about 20 kDa. Treatment of melanoma cells with 5 U/ml of phosphatidylinositol‐specific phospholipase C completely abolished cell‐surface expression of CD59. Interferon‐γ and/or tumor necrosis factor‐α or phorbol 12‐myristate 13‐acetate neither modulated the expression of CD59 by melanoma cells nor influenced the amounts of CD59‐specific mRNA. F(ab')2 fragments of anti‐CD59 MAb YTH53.1 did not inhibit the lysis of melanoma cells by allogeneic natural killer (NK) cells or lymphokine‐activated killer (LAK) cells. In contrast, the whole lg molecule of MAb H19 or YTH53. I significantly (p < 0.05) enhanced NK‐cell‐mediated lysis of melanoma cells, suggesting the induction of antibody‐dependent cell‐mediated cytotoxicity. Lastly, masking of CD59 by MAb YTH53. I or its F(ab')2 fragments significantly (p < 0.05) enhanced, in a dose‐dependent fashion, the lysis of anti‐GD3‐sensitized melanoma cells by homologous complement. These data demonstrate that CD59 expressed by human melanoma cells might regulate host‐tumor interaction by protecting neoplastic cells from complement‐mediated lysis. © 1995 Wiley‐Liss, Inc.
Quantitative and qualitative effects of experimental radioimmunotherapy on tumor vascular permeability
Authors: Blumenthal Rosalyn D. , Kashi Rina , Sharkey Robert M. , Goldenberg David M.
Published: February 2007

Localization of radiolabeled antibodies in the perivascular space of tumors resulted in morphological changes in blood vessel structure and physiological changes in tumor vessel function. Vessel diameter decreased by day 14 and was associated with a significant decline in vascular volume (VV). Upon recovery of VV, the basement membrane surrounding the endothelium had thickened. Tumor vascular permeability (VP) decreased within 7 days of treatment and remained suppressed throughout the 42‐day observation period of our study. The decline in VP, which could be visualized by fluorescent microscopy of FITC‐dextran extravasation, was dose‐related and could be quantified at doses as low as 800 cGy. The radioantibody‐induced 50–80% decline in tumor VP was observed in 3 human colonic xenografts (GW‐39, LS174T and MOSER). Decreases in VP have been observed for proteins ranging in size from 20 to 150 kDa. Similar effects on VP were noted when low protein doses (10–30 μg), which resulted in heterogeneous antibody distribution, were used or with high protein doses (400–750 μg), which resulted in more uniform penetration of antibody. If 188Re or 90Y, radiometals with higher β‐energy and longer path lengths, were substituted for 131I, a similar decrease in VP was observed. The radioimmunotherapy (RAIT)‐induced decrease in tumor VP resulted in a 90% decline in accretion of a second dose of radioantibody. The 10% of the second dose that was taken up by the tumor targeted many already non‐viable tumor regions and some, but not all, viable tumor cell clusters. © 1995 Wiley‐Liss, Inc.
Calibration of misonidazole labeling by simultaneous measurement of oxygen tension and labeling density in multicellular spheroids
Authors: Gross Markus W. , Karbach Ulrich , Groebe Karlfried , Franko Allan J. , Mueller‐Klieser Wolfgang
Published: February 2007

To correlate misonidazole concentrations and oxygen pressures (Po2) at identical locations within EMT6/Ro multi‐cell spheroids (mean diameters ± SD: 867 ± 20 μm), Po2 measurements were performed with oxygen‐sensitive microelectrodes during incubation of these spheroids with tritiated misonidazole (10 mg/l; 445 μCi/mg). In each individual spheroid, Po2 profiles were correlated with the corresponding spatial distribution of misonidazole as quantified by conventional autoradiography and grain counting. To compare the oxygenation status of spheroids in the measuring chamber with that of spheroids in spinner culture, misonidazole labeling was performed in both environments following the same protocol. All experiments were conducted in 20% oxygen and BME or in 5% oxygen and DMEM to obtain spheroids with different degrees of oxygenation. Labeled misonidazole was fairly evenly distributed in the outer, better oxygenated regions of EMT6 spheroids. In contrast, there was an accumulation of the labeled substance near central necrosis where low oxygen tensions were measured. Grain densities were similar at corresponding oxygen pressures under both environmental conditions. Except for some scatter, grain density as a function of oxygen pressure showed little variation in the Po2 range of 20–60 mm Hg, but exhibited a steep increase below 10 mm Hg. The findings imply that a substantial rise in local misonidazole labeling indicates a meta‐bolically active tissue region at low Po2 that is not necessarily identical with the radiobiologically hypoxic cell fraction. A comparison of the labeling densities of spheroids in spinner flasks and in the Po2 measuring chamber indicates that oxygenation of spheroids is better in rotation culture than during microelectrode measurements. © 1995 Wiley‐Liss, Inc.
B‐cell lymphomagenesis in SIV‐immunosuppressed cynomolgus monkeys
Authors: Rezikyan Susanna , Kaaya Ephata E. , Ekman Marianne , Voevodin Alexander F. , Feichtinger Hans , Putkonen Per , Castaños‐Velez Esmeralda , Biberfeld Gunnel , Biberfeld Peter
Published: February 2007

B‐cell lymphomas developed frequently (approx. 40%) in SIVsm (SMM3) immunosuppressed monkeys and were mostly extranodal, aggressive and all associated with an EBV‐related simian herpes virus operationally designated herpes virus Macoca fascicularis (HVMF‐1). Lymphoma tissues from 21 monkeys were studied by PCR and DNA PAGE for mono/oligoclonality of the VDJ‐rearranged IgH genes. Most lymphomas (n = 15) showed a monoclonal and approximately 1/3 (n = 6) an oligoclonal VDJ rearrangement pattern. The time after infection to tumor presentation was significantly shorter for oligoclonal than for monoclonal lymphomas, suggesting that oligoclonal selection frequently precedes the outgrowth of a single malignant clone. Comparison of the VDJ rearrangements in an established lymphoma cell line and the original, oligoclonal lymphoma tissue indicated in vitro selection of one HVMF‐infected clone. Longitudinal studies of sequential lymph‐node biopsies showed that the malignant lymphoma clone in 3 out of 8 lymphomas could be identified as a predominant clone in lymph nodes 2‐12 months after SIV infection and 6‐10 months before clinical presentation of the lymphomas. VDJ‐rearranged DNA corresponding to that of the lymphomas was also detected in most sera at the time of lymphoma manifestation but not in corresponding PBL preparations. Clearly, the SIVsm AIDS model in cynomolgus monkeys represents a powerful tool for biological and clinical studies of herpes‐virus‐associated lymphomagenesis in immunosuppressed states. © 1995 Wiley‐Liss, Inc.
International renal‐cell cancer study. IV. Occupation
Authors: Mandel Jack S. , McLaughlin Joseph K. , Schlehofer Brigitte , Mellemgaard Anders , Helmert Uwe , Lindblad Per , McCredie Margaret , Adami Hans‐Olov
Published: February 2007

The relationship between renal‐cell cancer (RCC) and occupation was investigated in an international multicenter population‐based case‐control study. Study centers in Australia, Denmark, Germany, Sweden and the United States interviewed 1732 incident RCC cases and 2309 controls. Significant associations were found with employment in the blast‐furnace or the coke‐oven industry [relative risk (RR), 1.7; 95% confidence interval (CI), 1.1–2.7], the iron and steel industry (RR, 1.6; 95% CI, 1.2–2.2) and exposure to asbestos (RR, 1.4; 95% CI, 1.1–1.8), cadmium (RR, 2.0; 95% CI, 1.0–3.9), dry‐cleaning solvents (RR, 1.4; 95% CI, 1.1–1.7), gasoline (RR, 1.6; 95% CI, 1.2–2.0) and other petroleum products (RR, 1.6; 95% CI, 1.3–2.1). Asbestos, petroleum products and dry‐cleaning solvents appear to merit further investigation, in view of the relationship between risk and duration of employment or exposure and after adjustment for confounding. There was a negative association between RCC and education, but it was not consistent across all centers. Overall, the results of our multicenter case‐control study suggest that occupation may be more important in the etiology of RCC than indicated by earlier studies. © 1995 Wiley‐Liss, Inc.
Is residence in areas of endemic goiter a risk factor for thyroid cancer?
Authors: Galanti Maria Rosaria , Sparén PÄR , Karlsson Anders , Grimelius Lars , Ekbom Anders
Published: February 2007

The association between an indicator of iodine deficiency and differentiated thyroid cancer has been investigated in a population‐based case‐control study. This included all incident cases (N = 484) of papillary and follicular thyroid cancer diagnosed during the years 1980–1992 in a Swedish Health Care Region among residents born in Sweden, and as many individually matched controls. Cases were included after a uniform review of their histopathological specimens. Residence in areas where goiter had been severely endemic in the 1930s was used as exposure indicator. Odds ratios (OR) and 95% confidence intervals as estimates of relative risk were calculated as a measure of association using logistic regression. A trend toward an association was found with a duration of residence in goiter areas between 21 and 40 years, most prominent among follicular cancer cases and in the group diagnosed at age ≶ 50 years compared to not exposed. Exposure for the first time during adolescence (between 11 and 20 years) was associated with an increased risk of papillary cancer. This was especially evident among women, both when compared to not exposed and to those exposed during the first year of life; the association was strengthened after adjustment for duration of residence. These findings support the hypothesis of distinct causation patterns from iodine deficiency to the 2 most common histological types of thyroid cancer. © 1995 Wiley‐Liss, Inc.
Time trend in pancreatic cancer incidence in connecticut, 1935–1990
Authors: Zheng Tongzhang , Holford Theodore R. , Ward Barbara A. , McKay Lisa , Flannery John , Boyle Peter
Published: February 2007

A total of 13,246 incident pancreatic cancer cases, reported to the Connecticut Tumor Registry between 1935 and 1990, were included in our study. Results indicate that the overall age‐adjusted incidence rate of pancreatic cancer increased between 1935 and 1964 in males, and leveled off thereafter. Since 1975, the incidence rate has in fact been decreasing, from 12.04/100,000 in 1975–79 to 10.44/100,000 in 1985–90. In females, the overall age‐adjusted incidence rate also increased between 1935 and 1974. Since then, however, it has remained relatively stable. Age‐specific incidence rates also show no signs of an increase in rate from any age group, or from either sex for recent birth cohorts. There is also no clear increase in the incidence of pancreatic cancer in any of the ethnic/gender categories since the early 1970s. Age‐period‐cohort modeling shows that the increasing birth cohort trend peaked among those born around 1920–1925 and, for recent birth cohort, a slightly decreasing trend was observed in both males and females. Our study reveals no signs of an increase in the immediate future in pancreatic cancer incidence rate in any of the sex, ethnic and age groups in Connecticut. © 1995 Wiley‐Liss, Inc.
Mechanism of interaction between cisplatin and human recombinant interferon gamma in human ovarian‐cancer cell lines
Authors: Nehmé Alissar , Albin Nicolas , Caliaro Marie Josée , Guichard Sylvie , Jozan Suzanne , Julia Anne‐Marie , Bugat Roland , Canal Pierre
Published: February 2007

Human ovarian carcinoma cells (2008 and its cisplatin‐resistant sub‐line 2008/C13*) were sensitized to cisplatin by treatment with human recombinant gamma interferon (IFNγ). IFNγ produced no significant change in the uptake of CDDP. Exposure of 2008 and 2008/C13* cells to IFNγ resulted in a time‐dependent decrease of cellular glutathione and total glutathione‐S‐transferase activity, principally the π isoform. By contrast, the treatment of 2008 and 2008/C13* cell lines with IFNγ induced rather than suppressed metallothionein IIA mRNA levels. IFNγ changed neither the formation of total platinum‐DNA adducts, nor DNA repair. A significant decrease in c‐erbB‐2 expression was observed both in sensitive and in resistant cell lines after treatment with IFNγ, and this decrease was dose‐dependent. Our results indicate that the mechanism of IFNγ‐induced sensitization in human ovarian‐cancer cell lines is multifactorial. © 1995 Wiley‐Liss, Inc.
RB‐reconstituted human retinoblastoma cells form RB‐positive intraocular and intracerebral but not subcutaneous tumors in scid mice
Authors: Szekely Laszlo , Wang Yisong , Klein George , Wiman Klas G.
Published: February 2007

WERI‐Rb 27 human retinoblastoma cells were reconstituted with an intact RB gene by retrovirus‐mediated gene transfer, in order to study the phenotypic effects of the protein in vitro and in vivo. Extensive morphological changes were observed, dominated by the formation of multinucleated giant cells. Six weeks after retroviral infection, the giant cells began to die and small cells emerged, resembling the parental non‐reconstituted line. They expressed RB and continued to grow, although they showed an increased sensitivity to serum starvation. The original RB‐negative cells grew progressively after subcutaneous inoculation into SCID mice, whereas the reconstituted cells failed to grow. RB‐positive cells grew progressively in the corpus vitreum of the eye and in the brain, however. The RB‐reconstituted cells grew more slowly and were less invasive than the parental cells and cells infected with a firefly luciferase (LUX) gene carrying retrovirus, used as controls. RB‐reconstituted cells re‐explanted from the intraocular and intracranial tumors continued to express full‐length RB protein. RBeye2, an RB‐positive cell line established from an eye tumor, was still unable to grow subcutaneously. The reduced tumorigenicity of the RB‐reconstituted cells in the subcutaneous space may be due to the influence of locally acting growth‐controlling signals or the absence of microenvironment‐specific trophic factors. Alternatively, it may reflect the action of residual immune effectors in the SCID mice. If this is the case, these would have to be more effective at the subcutaneous site than in the eye or brain. © 1995 Wiley‐Liss, Inc.
Alterations in the signal‐transducing molecules of T cells and nk cells in colorectal tumor‐infiltrating, gut mucosal and peripheral lymphocytes: Correlation with the stage of the disease
Authors: Matsuda Masanori , Petersson Max , Lenkei Rodica , Taupin Jean‐Luc , Magnusson Inger , Mellstedt Håkan , Anderson Paul , Kiessling Rolf
Published: February 2007

T cells from mice bearing an experimental colon carcinoma, and from patients with colorectal and renal carcinomas, have atypical T‐cell receptors (TCR). In the present study, further characterization of modulations in CD3‐ and CD 16‐associated ζ chain in peripheral blood lymphocytes (PBL) and tumor‐infiltrating lymphocytes (TIL) from colorectal carcinomas was performed. Relative to PBL, the percentage of natural killer (NK) cells among fresh TIL was reduced, while a higher proportion of T cells expressing HLA‐DR was found. As previously reported, we found significantly reduced levels of the CD3‐ and CD 16‐associated ζ chain in TIL and, to a lesser extent, also in patients' PBL. Levels of ζ chain in T and NK cells from non‐cancerous colorectal tissue from patients were lower than in PBL but higher than in TIL, with a direct relationship between levels of this signal‐transducing molecule and the distance from the tumor. In addition, ζ levels correlated with the Dukes' stage of the disease, since PBL from patients with lymph‐node involvement or distant organ metastases (Dukes' stages C and D) had significantly less CD3 ζ than patients with localized disease (stages A and B). Patients' T cells also had decreased levels of cell‐surface and cytoplasmic CD3 ϵ. We also observed reduced levels of the TCR accessory molecules CD4 and CD8, mainly on TIL but to a lesser extent also on patients' PBL. Biochemical analysis of anti‐CD3 ϵ‐immunoprecipitated TCR complexes demonstrated that the CD3 complex was not associated with the ζ chain, either on TIL or on PBL or on lymphocytes from non‐cancerous colon tissue, suggesting a defect in the assembly of the TCR complex. Following several days of in vitro culture with recombinant interleukin‐2 and phytohemagglutinin, anti‐CD3 or anti‐CD2 monoclonal antibodies (MAbs), levels of CD3 ζ chain as well as of cell surface CD3 ϵ were normalized. Our findings suggest an abnormal expression as well as assembly of several different signal‐transducing molecules of T cells and NK cells, which correlate with the stage of the disease in patients with colorectal carcinomas. © 1995 Wiley‐Liss, Inc.