bluuebunny/crossref_metadata_2025_split huggingface dataset:
Isolation and characterization of tumor‐infiltrating lymphocytes from cervical carcinoma
Authors: Hilders Carina G. J. M. , Ras Liesbeth , van Eendenburg Jaap D. H. , Nooyen Yvonne , Fleuren Gert Jan 
Published: February 2007

AbstractThe evidence that virus‐induced tumors generally elicit T‐cell responses prompts the notion that HPV‐related cervical carcinoma would be amenable to treatment by T‐cell‐mediated adoptive therapy. Therefore, we cultured and cloned tumorinfiltrating lymphocytes (TIL) from a patient with cervical carcinoma and studied the in vitro characteristics of these TIL by using the established autologous tumor‐cell line. After stimulation of bulk TIL cultures with 1,000 Units/ml recombinant interleukin 2 (rIL‐2), followed by limiting dilution, T‐cell clones were generated in the presence of 20 U/ml rIL‐2 and irradiated autologous tumor cells, PBLs and EBV‐transformed B‐cell lines. Phenotypically, all clones were CD3/CD8‐positive with a heterogeneous CD56 expression. All expressed preferential cytolytic activity against autologous tumor cells, did not lyse autologous lymphoblasts, and were cytotoxic against the NK‐sensitive cell line K562. A minor lytic capacity was detectable on allogeneic cervical tumor‐cell lines or tumor‐cell lines of other histologic types. Cytotoxicity against the autologous tumor could be inhibited by anti‐CD3, anti‐CD8 and anti‐ICAMI but not by anti‐HLA class‐1 (W6/32, B9.12.I), anti‐allele‐specific HLA determinants and anti‐LFA‐3 antibodies. We demonstrate a highly specific autologous lytic activity of cervical carcinoma TIL, in which a CD3‐associated surface antigen recognition is involved. These results may prove useful in further studies on adoptive immunotherapy of cervical cancer patients. © 1994 Wiley‐Liss, Inc.
Cell kinetics in human breast cancer: Comparison between the prognostic value of the cytofluorimetric S‐phase fraction and that of the antibodies to Ki‐67 and PCNA antigens detected by immunocytochemistry
Authors: Gasparini Giampietro , Boracchi Patrizia , Verderio Paolo , Bevilacqua Pierantonio 
Published: February 2007

AbstractThe determination of cell proliferation is one of the more widely used tools for assessing prognosis. However, additional research in this field is warranted because today there are several methodological procedures available for monitoring cell kinetics and it has still not been established which is the most reliable marker of proliferation and which possesses the greatest prognostic value. We performed this study in a series of primary invasive breast cancers to compare the prognostic value of S‐phase fraction (SPF) by flow cytometry, the most widely used method for detecting proliferation at present, with that of antibodies to Ki‐67 and PC‐10 to proliferating‐cell nuclear antigen (PCNA) detected by immunocytochemical methods. A significant linear relationship was observed only between SPF and Ki‐67. In univariate analysis SPF and Ki‐67 values, nodal status, histological grading and peritumoral lymphatic‐vessel invasion were significant predictors of relapse‐free survival (RFS). As far as overall survival (OS) is concerned, only SPF, Ki‐67 and nodal status were significantly associated with the risk of death. PCNA had no prognostic value for either RFS or OS. In multivariate analysis only SPF and nodal status retained a significant and independent prognostic value. Neither the cell‐kinetics parameters assessed by immunocytochemistry (i. e. Ki‐67 and PCNA) nor histological grading were independent prognosticators. In conclusion, our results provide evidence that the determination of SPF by flow cytometry was the strongest cell‐kinetics marker used to assess prognosis in this series of breast cancers. However, different and novel markers of cell kinetics need to be compared in larger series in order to identify the best one. © 1994 Wiley‐Liss, Inc.
Phorbol ester induced rapid attachment and spreading of melanoma cells and the role of extracellular matrix proteins
Authors: van Leeuwen Robert L. , Dekker Sybren K. , Arbiser Jack L. , Vermeer Bert Jan , Bruijn Jan A. , Byers H. Randolph 
Published: February 2007

AbstractPhorbol 12‐myristate 13‐acetate (PMA) is a tumour promotor that acts as a potent protein kinase C (PKC) activator that has significant effects on tumour cell attachment and spreading. We tested whether these effects of PMA may be observed in human melanoma cells, and whether a specific response to extracellular matrix proteins may be mediated by shifts in the expression of β1 integrins. We used cell attachment assays, video time lapse cell spreading assays, flow cytometry, function blocking monoclonal antibodies (MAbs) and PKC inhibitor Calphostin C to address these questions. We established that PMA induces a rapid and temporary enhancement of cell attachment and spreading which was not accompanied by a significant change in the expression of β1 integrins. Spreading of melanoma cells that were not stimulated with PMA could be significantly blocked with a function blocking MAb (clone P4C10) against the common β1 integrin subunit. The spreading and attachment of the PMA treated cells was also significantly reduced, but less so, after MAb treatment. The PMA enhanced cell attachment and spreading could be effectively blocked by RGD sequences and PKC inhibitor. Taken together, our data indicate that PMA induces a rapid and temporary ECM‐dependent enhancement of melanoma cell attachment and spreading, and that the response to ECM components appears not to be due to significant shifts in β1 integrin expression, but rather to activation of β1 integrins. © 1994 Wiley‐Liss, Inc.
E‐selectin binding by pancreatic tumor cells is inhibited by cancer sera
Authors: Sawada Tetsuji , Ho Jenny J. L. , Chung Yong‐Suk , Sowa Michio , Kim Young S. 
Published: February 2007

AbstractTumor cells interact with endothelial cells during both intraand extravasation. Understanding how these interactions are modulated could lead to the development of ways to alter the metastatic potential of tumor cell. Three pancreatic cancer cell lines, SW1990, CAPAN‐2 and PANC‐1, were examined for their ability to bind to the endothelial cell adhesion molecule E‐selectin (ELAM‐1). SW1990 cells exhibited highest binding, highest surface expression of the carbohydrate antigens sialylated Lewis2 (sLe2) and sialylated Lewisx (sLex) and released the most high m. w. sLe2 and sLex antigens. Expression of sLe2 and sLex antigens and binding to E‐selectin were reduced by pre‐treatment of SW1990 cells with the O‐linked glycosylation inhibitor benzyl‐α‐GaINAc but not with the N‐linked glycosylation inhibitor tunicamycin. Expression of peptide epitopes associated with MUCI apomucins was increased by benzyl‐α‐GaINAc. Cell binding was greatly reduced by mucins purified from SW1990 xenografts and by an antibody against sLe2. An antibody against sLex had a much less marked effect. Sera from pancreatic cancer patients reduced SW1990 cell binding to E‐selectin but sera from normals did not. The degree of inhibition was related to the sLex level in the sample. When cancer serum was separated by column chromatography on Sephacryl S‐400, the void volume fractions contained most of the sLe2 and sLex antigens and most of the inhibitory activity to E‐selectin binding. Differences in the relative availability of sLe2 and sLex ligands on serum molecules and on the SW1990 cell surface may account for the differences between antibody and serum inhibition results. Thus SW1990 cell adhesion to E‐selectin is mediated by ligands on mucinous glycoproteins, and adhesion can be inhibited by mucins, high blood levels of sLex and reduction of cellular O‐linked glycosylation. © 1994 Wiley‐Liss, Inc.
Expression of stromelysin‐3 and nm23 in breast carcinoma and related tissues
Authors: Hähnel Erika , Dawkins Hugh , Robbins Peter , Hähnel Roland 
Published: February 2007

AbstractBiological functions proposed for the ST3 and mm23 genes in tumour development and progression seem to be directly opposed. Stromelysin‐3 (ST3) is a putative member of the matrix metalloproteinase family. ST3 has been implicated in the progression of epithelial malignancies, specifically with regard to an invasive (and therefore potentially metastasizing) phenotype. The nm23 gene, on the other hand, encodes a nucleoside diphosphate kinase which allegedly has a metastasis‐suppressor‐type function. It was therefore of interest to compare the expression of ST3 and nm23 in various surgically excised normal and neoplastic breast tissues. RNA was isolated from over 200 surgical specimens and studied by Northern blots. Normal breast tissues did not express ST3, and ST3 expression was detected in only 1 of 20 normal axillary lymph nodes. None of 7 fibroadenomas expressed ST3. In contrast, 60% of primary and metastatic breast carcinomas contained ST3‐mRNA. The expression of ST3 was mainly confined to invasive carcinomas and was observed less frequently in pure ductal carcinoma in situ (DCIS) lesions. Our results support the suggestion that ST3 expression is related to the malignant process in breast cancer. The role of nm23 is far less clear‐cut.
Simple tandem repeat allelic deletions confirm the preferential loss of distal chromosome 6q in melanoma
Authors: Walker Graeme J. , Palmer Jane M. , Walters Marilyn K. , Nancarrow Derek J. , Parsons Peter G. , Hayward Nicholas K. 
Published: February 2007

AbstractKaryotypic analysis, loss of somatic heterozygosity, microcell fusion and cDNA transfection studies have provided compelling evidence that at least one tumour suppressor gene for melanoma resides on chromosome 6. In an attempt to further define the regions to which these putative suppressor genes map, we have carried out loss of heterozygosity (LOH) studies on DNA from 25 fresh melanoma tumours for 9 simple tandem repeat (STR) polymorphism markers spanning chromosome 6. Four samples displayed LOH or homozygosity for all markers studied, indicating that they had lost one homologue of chromosome 6. An additional 3 samples showed LOH for all markers on 6q. Furthermore, 30 melanoma cell lines, for which there were no matching somatic DNA samples, were analyzed for hemizygosity of markers on 6q. One cell line had a homozygous deletion of all markers tested and a further 12 cell lines displayed only one allele for 3 or 4 contiguous markers, indicating that most, if not all of these samples were hemizygous for the region of 6q distal to D6S87. Overall, the rate of LOH on 6q in the 55 melanoma DNAs was 35%, and there were no losses of markers on 6p without concomitant loss of markers on 6q. Two of 5 samples derived from primary melanomas showed LOH, which indicates that LOH for the melanoma suppressor gene on 6q, which maps to a region that contains the SOD2 locus, is a frequent and early event in melanoma tumorigenesis.
Prognostic value of deoxyribonucleic acid content in prostate cancer: A review of current results
Authors: Adolfsson Jan 
Published: February 2007

AbstractA total of 115 articles on prostate cancer were reviewed for data on the prognostic value of DNA content in the tumor cells. In 44 series, data pertinent to this review were found. There was no consensus in the literature with respect to methods of analysis of DNA content or definitions of subclasses of DNA content such as categories of ploidy. The DNA content of prostate cancer cells was strongly related to tumor grade and stage. When analyzed as a single parameter in univariate analyses, the DNA content had a prognostic value with respect to overall or disease‐specific survival. In multivariate analyses the additional prognostic value of the DNA content was less convincing when analysed with tumor grade and stage. The prognostic data from univariate and multivariate analyses available in the literature were mainly derived from patients with advanced disease and data on localized, potentially curable disease were scanty and conflicting.
Stimulation of angiogenesis as an explanation of matrigel‐enhanced tumorigenicity
Authors: Daniel Bonfil R. , Vinyals Antonia , Bustuoabad Oscar D. , Llorens Ana , Benavides Fernando J. , Gonzalez‐Garrigues Milagro , Fabra Angels 
Published: February 2007

AbstractMatrigel, a reconstituted extract of basement membrane, enhances the growth of different human cancer cell lines when transplanted into nude mice. Here that stimulation was confirmed in the BALB/c murine mammary‐tumor cell line M3MC, as well as in human colon (SW948) and mammary (MDA‐MB‐468) carcinoma cell lines transplanted in nude and SCID mice, respectively. Subcutaneous and intra‐mammary fat‐pad inoculations of Matrigel alone generated an angiogenic response which was macroscopically evident by day 9. Histological analysis of the local host reaction occurring at the site of injection revealed an early peripheral fibroblast response, followed by mononuclear cell infiltration, solid and hollow fibroblast cords projections from the edge to the center of the Matrigel plug, and finally capillary ingrowths. Conditioned media obtained from the gels generated in vivo, acted as very strong chemoattractants for mouse lung capillary endothelial cells, stimulating their motility between 38 and 82 times with respect to the control. Our results suggest an important role of host cells recruited by Matrigel, which could favor angiogenesis of the area and thus facilitate the growth of tumor cells co‐inoculated with the basement membrane extract.
Generation of monoclonal antibodies specific for ganglioside lactones: Evidence of the expression of lactone on human melanoma cells
Authors: Kawashima Ikuo , Kotani Masaharu , Ozawa Hideki , Suzuki Minoru , Tai Tadashi 
Published: February 2007

AbstractWe generated 3 murine monoclonal antibodies (MAbs) specific for ganglioside lactones by immunizing C3H/HeN mice with purified lactones adsorbed to Salmonella minnesota followed by fusion with mouse myeloma cells. The use of a wide variety of glycolipids, including ganglioside lactones, enabled the precise structures recognized by these MAbs to be elucidated through an ELISA and by immunostaining on thin‐layer chromatography. MAb AMR38, which was generated with GMI lactone, showed restricted specificity, detecting only the GMI lactone used for immunization. None of the other ganglioside lactones, intact gangliosides (including GMI) or neutral glycolipids tested were recognized. In contrast, MAbs AMR40 and AMR19, which were generated with GD Ia lactone and GD3 lactone, respectively, showed broader specificities, recognizing several ganglioside lactones. However, the precise epitopes were different. MAb AMR40 reacted intensely with ganglioside lactones having an external NeuAcα2 → 3Gal‐ sequence (GD Ia, GM3, GM Ib, GT Ib, and IV3Neu´‐nLC4Cer), but not with those having a NeuAcα2 → 8NeuAcα2 → 3Gal‐ sequence. On the other hand, MAb AMR 19 reacted with ganglioside lactones having a NeuAcα2 → 8NeuAcα2 → 3Gal‐ sequence (GD3, O‐Ac‐GD3, GD2, GD Ib, GT Ib, GQ Ib and GT Ia), but not with those having a NeuAcα2 →3Gal‐ sequence. None of the intact gangliosides or neutral glycolipids tested were recognized by the MAbs. We also determined the expression of ganglioside lactones on human melanoma cells grown in athymic nude mice by means of an immunofluorescence technique.
A nonconsensus octamer‐recognition sequence (TAATGARAT‐motif) identifies a novel DNA binding protein in human merkel cell carcinoma cell lines
Authors: Thomson J. A. F. , Leonard J. H. , McGregor K. , Sturm R. A. , Parsons P. G. 
Published: February 2007

AbstractWe have established a number of cell lines from Merkel cell carcinoma (MCC) of the skin. In many respects these cell lines resemble those established from small cell lung cancer (SCLC) and it is difficult to differentiate between metastatic MCC and SCLC on morphological or histochemical criteria. Both are thought to be neuroendocrine tumours and may express a number of neuroendocrine markers. SCLC cell lines express the octamer‐DNA binding transcription factor brn‐2 gene products N‐Oct 3 and N‐Oct 5, which are restricted to the neuroectodermal cell lineage. In DNA binding studies using a consensus octamer recognition site we have found that 4 of 8 MCC cell lines examined expressed brn‐2 in at least trace amounts compared with 3 SCLC cell lines which all expressed brn‐2 proteins at high levels. Moreover, these DNA binding studies were extended by using a high‐affinity brn‐2 recognition site related to the degenerate octamer TAATGARAT‐motif. This identified a novel DNA binding protein in a subset of MCC cell lines. The protein was absent from the three SCLC cell lines, melanoma cells and brain tissue. This binding activity, which we term Merkel cell DNA nuclear factor (MNF), was shown to be specific by competitive inhibition with oligonucleotide binding sites and was not inhibited by polyclonal antisera against the Oct‐I, Oct‐2 or Brn‐2 proteins. This protein may serve as a unique marker for MCC compared with SCLC cells and may be involved in regulating the Merket cell phenotype.
In vivo evidence of the role of α4β1‐VCAM‐1 interaction in sarcoma, but not in carcinoma extravasation
Authors: Paavonen Timo , Tiisala Sinikka , Majuri Marja‐Leena , Böhling Tom , Renkonen Risto 
Published: February 2007

AbstractTumor‐cell invasion can occur via either lymphatics or blood vessels. When in the blood circulation, tumor cells have to adhere to endothelium lining the blood vessels before they can extravasate. Several families of adhesion molecules have been recognized: selectins and their oligosaccharide‐containing ligands and integrins and their counter‐receptors belonging to the immunoglobulin superfamily. Besides their essential role in leukocyte extravasation, these adhesion molecules have been proposed by in vitro experiments to be involved in tumor‐cell invasion by facilitating the adhesion of malignant cells to endothelium leading to extravasation and metastasis. We have previously shown that, in vitro, several sarcoma cell lines adhere strongly to cultured endothelial cells via α4β1‐VCAM‐1 interaction. Here we show that sarcoma cells, especially in the metastatic lesions, were strongly α4β1 positive but did not express α4β7, which is another receptor for VCAM‐1. Furthermore, we demonstrate that the capillary endothelium within metastatic sarcoma lesions reacted strongly with anti‐VCAM‐1 antibody and very often the α4β1‐expressing sarcoma cells were localized in the close vicinity of VCAM‐1‐expressing vessels. As control material we analyzed carcinoma specimens, but could not detect any α4‐integrin expression on malignant cells even though the endothelial cells were often VCAM‐1 positive. These results suggest that carcinomas do not use α4β1‐VCAM‐1 in extravasation and, taken together, provide circumstantial evidence that in vitro findings of α4β1‐VCAM‐1‐dependent sarcoma cell adhesion to endothelium can be extended to in vivo situations.
Frequency of cytotoxic T lymphocyte precursors (CTLp) interacting with autologous tumor via the T‐cell receptor: Limiting dilution analysis of specific CTLp in peripheral blood and tumor‐invaded lymph nodes of melanoma patients
Authors: Mazzocchi Arabella , Belli Filiberto , Mascheroni Luigi , Vegetti Claudia , Parmiani Giorgio , Anichini Andrea 
Published: February 2007

AbstractThe frequencies of cytotoxic T‐lymphocyte precursors (CTLp) that lyse auto logo us tumor by a T‐cell receptor (TCR) dependent mechanism (specific CTLp) were evaluated by limiting dilution analysis (LDA) using lymphocytes from peripheral blood (PBL) and from surgically resected, tumor‐invaded lymph nodes (LNL) in 9 melanoma patients. The frequency of specific CTLp was determined in PBLs and/or LNIs of all patients by a modified LDA assay, enabling us to measure lytic activity on the autologous tumor that could be significantly inhibited by an anti‐CD3 monoclonal antibody (MAb). This assay allowed us to detect frequencies of specific CTLp ranging from 1/720 to 1/32,037 in peripheral blood and from 1/328 to 1/22,061 in tumor‐invaded lymph nodes. These frequencies indicated that lymphoid populations from PBLs or LNLs of melanoma patients may contain as low as 30 to as much as 3,000 specific CTLp/106 lymphocytes. In addition, comparison of wells containing specific CTLp with those showing no inhibition by anti‐CD3 MAb indicated that specific CTLp represent between 3 and 88% of all precursors with lytic activity on the tumor. In 6 of 9 patients, no marked differences between PBLs and LNIs in specific CTLp frequencies were found. A 10‐fold increase of specific CTLp, in comparison to PBL and LNL, was found only in lymphocytes isolated from a subcutaneous metastasis of one patient. Our results indicate that CTLp interacting with autologous tumor by a TCR‐dependent mechanism exist in PBL and LNL of most melanoma patients, although a wide variation in their absolute number is evident among different patients.
High levels of transglutaminase expression in doxorubicin‐resistant human breast carcinoma cells
Authors: Mehta Kapil 
Published: February 2007

AbstractTissue type II transglutaminase (TGase) is a member of the TGase family that catalyzes Ca2+‐dependent covalent cross‐linking of several amines to the γ‐carboxamide group of protein‐bound glutamine residues. The degree of therapeutic efficacy or toxicity of drugs may be related to their ability to serve as a substrate for TGase and their covalent linkage to glutamine residues of regulatory proteins through the catalytic action of this enzyme. Here, doxorubicin (adriamycin)‐resistant human breast carcinoma MCF‐7ADR cells exhibited 40‐ to 6C‐fold higher TGase activity than control drug‐sensitive MCF‐7wt cells. The same was observed in vivo: a small proportion of tumor cells became positive for TGase after administration of adriamycin‐based chemotherapy to patients with breast carcinoma. Similarly, continuous culture of MCF‐7WT, cells in the presence of adriamycin led to the appearance of the drug‐resistant phenotype that was in turn associated with increased expression of TGase. This increase in TGase was specific for adriamycin resistance. Like most known TGases, MCF‐7ADR TGase was completely dependent on the presence of Ca2+ for its catalytic activity. Based on its immunoreactivity, the TGase in MCF.7ADR cells was identified as an 85‐kDa tissue‐type TGase and was present only in the soluble form. Immunoblot analysis revealed that the increase in TGase activity was due to accumulation of the protein. Two cytosolic proteins of approximately 20 and 30 kDa in MCF‐7 cells served as suitable acyl donor substrates in TGase‐catalyzed reactions.