bluuebunny/crossref_metadata_2025_split huggingface dataset:
Apoptotic cell death induced by a mouse‐human anti‐APO‐1 chimeric antibody leads to tumor regression
Authors: Coney Leslie R. , Daniel Peter T. , Sanborn David , Dhein Jens , Debatin Klaus‐Michael , Krammer Peter H. , Zurawski V. R. 
Published: February 2007

AbstractThe murine anti‐APO‐1 antibody (γ3, k) induces programmed cell death (apoptosis) following binding to the APO‐I antigen (m.w., 48 kDa) expressed, e.g., on activated or malignant lymphocytes. APO‐I expression on malignant cell lines and tissues suggested potential clinical utility supported by anti‐APO‐I‐mediated tumor regression in a nude mouse model. A mouse‐human anti‐APO‐1 chimeric antibody (γ3, k) with an affinity similar to that of the murine antibody was produced. Chimeric anti‐APO‐1 showed the same potential to inhibit growth of the SKW6.4 B‐lymphoblastoid cell line as murine anti‐APO‐1, In addition, both the chimeric and murine anti‐APO‐1 antibodies were equally capable of mediating complete macroscopic tumor regression of a SKW6.4 xenotransplant in SCID mice by induction of apoptosis. Induction of apoptosis was the only mechanism for tumor regression because neither murine nor chimeric anti‐APO‐1 showed anti‐tumor activity against solid H53 tumor (APO‐1 antigen‐positive, anti‐APO‐1 ‐resistant) xenotransplants. Our results indicate that the chimeric anti‐APO‐1 antibody effectively induces apoptosis and suggest that chimeric anti‐APO‐1 should be evaluated for the treatment of malignant cells expressing the APO‐1 antigen. However, chimeric anti‐APO‐1 might only be used therapeutically when the antibody can be targeted specifically to tumor cells.
Cloning of feline p53 tumor‐suppressor gene and its aberration in hematopoietic tumors
Authors: Okuda Masaru , Umeda Akiko , Sakai Tadashi , Ohashi Takashi , Momoi Yasuyuki , Youn Hwa‐Young , Watari Toshihiro , Goitsuka Ryo , Tsujtmoto Hajimc , Hasegawa Atsuhiko 
Published: February 2007

AbstractAlterations of the p53 tumor‐suppressor gene have been observed in a variety of human and mouse tumors. For investigation of the role of this gene in tumors of cats, feline p53 cDNA was molecularly cloned from a feline lymph‐node cDNA library. The cloned cDNA (FF53) contained the whole open reading frame of p53 gene encoding 386 amino acids. The amino‐acid sequence of the feline p53 gene showed 82.1% and 74.9% similarities with those of the human and mouse counterparts, respectively, and had structural characteristics in common with the p53 genes of several other species. Aberrations of the p53 gene were investigated by RT‐PCR and single‐strand conformation polymorphism analyses. Of 10 primary hematopoietic tumors and 3 lymphoma cell lines examined, one lymphoma and one lymphoma cell line had a point mutation of the p53 gene, resulting in single amino‐acid substitutions.
Correlation between stromelysin‐3 mRNA level and outcome of human breast cancer
Authors: Engel Georg , Heselmeyer Kerstin , Auer Gert , Bäckdahl Martin , Eriksson Elina , Under Stig 
Published: February 2007

AbstractThe expression level of strometysin‐3 (ST3) mRNA was analyzed by in situ hybridization of formalin‐fixed, paraffin‐embedded primary breast‐tumor samples from 76 patients. Digital image analysis of the dark‐field in situ hybridization signal was used to measure the maximal level of ST3 expression in each tumor. All 55 invasive ductal carcinomas and 9 of 10 invasive lobular carcinomas were positive for ST3. Invasive tumors had significantly higher levels of ST3 than in situ tumors. Furthermore, ST3 levels were higher in invasive ductal carcinomas than in invasive lobular carcinomas. The ST3 expression level was significantly correlated to fatal metastatic disease (mean follow‐up 104 months). ST3 levels of < 2,500 units were associated with distant metastasis in 46% of patients, whereas levels of >2,500 units were associated with metastasis in 79% of patients selected for study. ST3 mRNA levels did not correlate with tumor siie, microvessel density, DNA pioidy or estrogen‐receptor levels. Studies of ST3 expression may provide information valuable for the understanding of breast cancer biology and for prognosis.
Expression of an endogenous retroviral gene product in human placenta
Authors: Kitamura Masanori , Maruyama Naoki , Shirasawa Takuji , Nagasawa Ryuji , Watanabe Kazutada , Tateno Masatoshi , Yoshiki Takashi 
Published: February 2007

AbstractTo investigate the presence and potential pathophysiological role of endogenous retroviruses in humans, we prepared a recombinant protein using clone 4‐1, a proviral sequence. DNA fragments containing the env region of clone 4‐1 were subdoned into a prokaryotic expression vector (pET3), and 2 fusion proteins, SU413 and SU415, were then expressed in Escherichia coli after treatment with isopropyl‐β‐thiogalactopyranoside (1PTG). By sonicating lysates of the transformed E. coli, the recombinant protein SU4I3 was successfully separated from the native bacterial components, and was used to raise an antiserum in rabbits. In immunoblot analysis, this antiserum specifically recognized the recombinant protein, but did not react with other components of E. coli. This antiserum was then used for an immunofluorescence study of human placenta, in which the env gene transcript has been reported. As a result, the anti‐SU4l3 serum detected substances in syncytiotropho‐blasts and vascular endothelia from a human placenta. No such reactivity was detected in human kidney or human liver. Immunoblot analysis revealed that this antiserum reacted to a single molecule of 38‐kDa in placenta, and its reactivity was reduced by the antiserum absorbed with SU4I3 antigen. These findings suggest that human placental syncytiotrophoblasts and vascular endothelia preferentially express a molecule encoded by human endogenous retrovirus clone 4‐1.
Collagens, integrins and the mesenchymal drift in glioblastomas: A comparison of biopsy specimens, spheroid and early monolayer cultures
Authors: Paulus Werner , Huettner Claudia , Tonn Jorg C. 
Published: February 2007

AbstractTo analyze the process of mesenchymal differentiation in vitro, we examined 5 human glioblastomas as biopsy specimens, monolayer cultures and 3‐dimensional fragment spheroid cultures for the immunohistochemical expression of extracellular matrix (ECM) components (collagen types I, III ‐VI, laminin) and integrin receptors (β1, β2, β3 and β4 chains). mRNA for type‐I and type‐IV collagen at chains was quantified using reverse transcription‐polymerase chain reaction. In situ, glioma cells expressed β1, the common β chain of most integrin ECM receptors, while ECM components were restricted to vascular elements. Early monolayer cultures showed a marked increase in ECM components (interstitial collagen? more than basement membrane components), and coexpression of ECM components and glial fibrillary acidic protein (GFAP) by most cells, β2 and β3 integrins were upregulated in the primary cultures. In the fifth passages, GFAP‐positive cells were decreased and collagen‐expressing cells increased. The spheroids exhibited preserved GFAP staining, neoexpression of β4 integrin in some tumors, and variable ECM expression by glioma cells which was lower than that in monolayer cultures. ECM deposition usually commenced in central spheroid areas where the Ki‐67 proliferation index was low. We conclude that different culture systems are characterized by distinct expression patterns for ECM components and receptors, and that mesenchymal features in cultured gliomas arise due to transdifferentiation of glioma cells.
Possible involvement of multidrug‐resistance‐associated protein (MRP) gene expression in spontaneous drug resistance to vincristine, etoposide and adriamycin in human glioma cells
Authors: Hasegawa Shuji , Taniguchi Ken , Yokomizo Akira , Kuwano Takashi , Ono Mayumi , Mori Temaki , Hori Shigeaki , Kohno Kimitoshi , Kuwano Michihiko 
Published: February 2007

AbstractThe multidrug‐resistance phenotype in human tumors is partly associated with over‐expression of the 170 kDa‐P‐glycoprotein encoded by the multidrug‐resistance‐1 (MDRI) gene. Another related, but non‐P‐glycoprotein, multidrug‐resistance‐associated protein (MRP) gene encodes a 190 kDa membrane ATP‐binding protein. Glioblastoma multiforme is a highly malignant primary neoplasm of the central nervous system which is refractory to anti‐cancer chemotherapy, but the mechanism underlying this drug resistance is unknown. Out of glioma cell lines, 2, namely IN500 and T98G, which had elevated MRP mRNA levels, showed the highest resistance to multiple anti‐cancer agents such as etoposide, vincristine and adriamycin, and decreased intracellular accumulation of etoposide. In the remaining 5 cell lines, various degrees of sensitivity to adriamycin and etoposide appeared to correlate with their respective MRP mRNA levels. Our study proposes that MRP may be involved in spontaneous multidrug resistance in human gliomas.
S‐phase fraction after gating on epithelial cells predicts recurrence in node‐negative breast cancer
Authors: Wingren Sten , Stå Olle , Sullivan Siw , Brisfors Ann , Nordenskjöld Bo 
Published: February 2007

AbstractWe have compared the prediction of distant recurrence for S‐phase fraction (SPF) and DNA‐ploidy, as estimated by flow cytometry, on an epithelial cell population and an unselected cell population from 268 node‐negative breast‐cancer patients diagnosed between 1985 and 1988. The tumor tissue was mechanically disintegrated and divided for flow cytometric analysis using both gated cells containing cytokeratin 8 and 18 and ungated cells treated with a detergent‐trypsin solution. The relationship to distant recurrence was investigated for flow cytometric data, tumor size and estrogen and progesterone receptor content in univariate and multivariate Cox's regression analysis. The regression analyses were performed on 209 cases with S‐phase fractions estimated by both methods. In 11 cases, DNA‐ploidy classification differed, reflecting increased sensitivity to minor aneuploid peaks but a decreased ability to separate peaks in the near‐diploid region for the gated populations. When SPF were used in univariate analysis as a continuous parameter or the upper tertile was used as cut‐off value, SPF from the cytokeratin‐gated cell population were most closely associated with recurrence and contributed additional prognostic information to SPF from the unselected cell population in the multivariate analysis. Out of the following variables:tumor size, ER and PR status, SPF and DNA ploidy, only SPF from immunoselected cells contributed prognostic information in multivariate analysis. These results indicate that SPF from immunoselected cell populations improves the prediction of recurrence in node‐negative breast cancer.
Distribution of oncofetal fibronectin isoforms in normal, hyperplastic and neoplastic human breast tissues
Authors: Kaczmarek Janusz , Castellani Patrizia , Nicolo Guido , Spina Bruno , Allemanni Giorgio , Zardi Luciano 
Published: February 2007

AbstractTwo different oncofetal fibronectins (FN) have been reported: one, generated by O‐glycosylation in the splicing region IIICS that is recognized by monoclonal antibody (MAb) FDC‐6, and another, recognized by MAb BC‐I, generated by the alternative splicing of the FN pre‐mRNA which includes an extra type‐III repeat called ED‐B. Using these and 2 other MAbs (IST‐4 which recognizes all different FN isoforms and IST‐6 which recognizes only the FN molecules that do not include the ED‐B sequence) we have immunohistochemically studied 171 normal, hyperplastic and neoplastic breast‐tissue specimens. Although all normal specimens reacted strongly with MAbs IST‐4 and IST‐6, they did not show the presence of oncofetal FNs as established by the use of BC‐1 and FDC‐6. In contrast, out of the 97 cases of invasive ductal carcinomas studied, 90 (93%) and 96 (99%) reacted positively with BC‐I and FDC‐6, respectively, the reaction being observed in the tumoral stroma connective tissue and in tumoral vessels. Furthermore, invasive lobular carcinoma showed less intense and less frequent staining with BC‐1 and FDC‐6 (10 and 11 out of 14, respectively). We found differences in the distribution of the 2 oncofetal fibronectin isoforms within the same specimens. The most remarkable difference was observed in the tumoral vessels: in invasive ductal carcinoma MAb BC‐1 revealed a positive reaction with vessels in 78% of cases while FDC‐6 showed such a reaction in only 59% of cases.
ras‐family gene mutations in neoplasia of the ampulla of vater
Authors: Scarpa Aldo , Zamboni Giuseppe , Achille Alberto , Capelli Paola , Bogina Giuseppe , Iacono Calogero , Serio Giovanni , Accolla Roberto S. 
Published: February 2007

AbstractMutations in the first and second exons of Ha‐, Ki‐ and N‐ras oncogenes were investigated in 17 epithelial tumors of the ampulla of Vater by single‐strand conformation polymorphism analysis and direct sequencing of DNA fragments amplified by polymerase chain reaction. The panel included 12 intestinaltype adenocarcinomas, 3 villous adenomas, I papillary carcinoma and I neuroendocrine carcinoma. Six cases (35%) contained ras mutations, affecting codon 12 of Ki‐ras in 2 adenomas and 3 carcinomas, and of N‐ras in I adenoma. All mutations were found in adenomas and among cancers with adenomatous areas, whereas none of the cases lacking adenomatous areas contained mutations. This suggested that ampullary cancers represent heterogeneous diseases with respect to the presence or absence of adenomatous areas and, among those with adenomatous areas, with respect to the presence of activated ras genes. Ki‐ras mutated cases included 3 of 4 tumors which mainly involved the intraduodenal bile duct, thus suggesting that a proportion of Ki‐ras ‐mutated ampullary cancers might correspond to those originating from the epithelium of the bile duct component of the ampulla.
Loss of apc protein expressed by human colonic epithelial cells and the appearance of a specific low‐molecular‐weight form is associated with apoptosis IN vitro
Authors: Browne Sara J. , Williams Ann C. , Hague Angela , Butt Alison J. , Paraskeva Christos 
Published: February 2007

AbstractAPC (adenomatous polyposis coli) protein is differentially expressed in the normal colonic crypt and believed to be involved in colonic cell maturation. In this work we investigated whether expression of the APC protein is associated with cell death in colonic epithelial cells. We have previously reported an in vitro system to study apoptosis. Briefly, cells attached to the flask have a low frequency of apoptosis (1–3%), whereas cells that detach from the flask and float in the medium have a high proportion of apoptotic cells (36–96% depending on the cell line). The full‐length 300‐kDa or truncated APC protein, normally expressed by the attached cells (detected using the FE9 antibody), was found to be lost in the floating apoptotic cells in 8/11 colon tumour cell lines examined. In addition, the APC antibody FE9 detected a 90‐kDa protein in the floating apoptotic cells of all cell lines investigated, which was not present in attached cells. Furthermore, loss of full‐length APC and gain of the 90‐kDa protein was observed in the apoptotic cells of 2 cell lines derived from other tissues: the SV40‐transformed fibro‐blast cell line CMSV40fib and the lymphoblastoid B‐cell line BJA‐B. In cells repeatedly frozen and thawed, believed to induce necrotic cell death, full‐length or truncated APC was also lost, though a 95‐kDa protein distinct from that in apoptotic cells was observed. Specific loss of full‐length or truncated APC (resulting in a 90‐kDa protein in apoptotic cells but a 95‐kDa protein in necrotic cells) is therefore associated with cell death. Our findings suggest a possible role for APC in cell survival.
Inhibition of proliferation and induction of monocytic differentiation in HL60 human promyelocytic leukemia cells treated with bile acids In vitro
Authors: Zimber Amazia , Chedeville Annick , Gespach Christian , Abita Jean‐Pierre 
Published: February 2007

AbstractWe have tested the effect of several bile acids on the proliferation and differentiation of the HL60 human promyelocytic leukemia cell line in vitro. Deoxycholate, chenodeoxycholate and lithocholic acid caused dose‐dependent inhibition of cell proliferation and induction of differentiation along the monocyte/macrophage pathway as determined by morphology, NBT test, non‐specific esterase, and staining by monoclonal antibodies against specific cell‐surface antigens. Optimal effects were obtained at 100, 75, and 60 μM of the 3 bile acids respectively. Cell‐cycle flow‐cytometric analysis showed that a substantial fraction of HL60 cells accumulated at the G0/G1 transition. Protein‐kinase‐C inhibitors such as sphinganine and H‐7 inhibited the differentiation‐inducing effect of bile acids, suggesting a possible role for PKC in this regulation. When bile acids were combined with non‐effective concentrations of all‐trans retinoic acid, enhancement of the monocytic differentiation of THP‐1 human leukemia cells was observed. Our findings demonstrate induction of tumor‐cell differentiation by bile acids, compounds that present minimal undesirable effects in humans.
Ecto‐5'‐nucleotidase (CD73) in multidrug‐resistant cell lines generated by doxorubicin
Authors: Ujházy Peter , Klobušická Margita , Babušíková Olga , Strausbauch Paul , Mihich Enrico , Ehrke1 M. Jane 
Published: February 2007

AbstractCytochemical screening for a panel of enzymes revealed increased 5'nucleotidase (5'NT) expression in 3 of 3 Pglycoprotein 170 (Pgp 170)‐positive multidrug‐resistant (MDR) variants of the murine EL4 T‐lymphoma cell line (EL4/ADM, ER2 and ER13). Electron microscopic localization established the presence of the membrane‐bound ecto‐form of the enzyme. Nine other murine, human and Chinese hamster cell lines and their MDR variants were tested for ecto‐5'NT. Of these, 4 MDR variants (human cell lines MCF7A6, MCF7A2, HeLaJ2C and the murine cell line LI2I0A) showed increased expression of ecto‐5'NT, when compared with their parental cell lines. The findings with cells of human origin were confirmed by immunofluorescent localization with a specific monoclonal antibody (MAb) (27.2) against the human ecto‐5'NT. All MDR cell lines with elevated ecto‐5' NT expression were generated by doxorubicin treatment. These cells were more sensitive than their parental cell lines to AMP at concentrations of 1.5–3.0 mM, confirming that the expressed ecto‐5'NT was biologically active. The parental and MDR cells did not differ, in general, in their sensitivity to adenosine. An inhibitor of ecto‐5'NT, α,β‐methyleneadenosine 5'‐diphosphate, completely reversed the resistance of the EL4/ADM cell line to doxorubicin. The possibility exists of a functional relationship between the ecto‐5'NT molecule and the members of the ATP‐binding cassette transporter superfamily, important components of MDR, in some cell types.
Differential intracellular processing of the anthracycline drug ME2303 in doxorubicin‐sensitive (A2780) and ‐resistant (A2780AD) human ovarian cancer cells as studied with confocal laser scanning microscopy and image analysis
Authors: Huxham Ian M. , Barlow Andrew L. , Lewis Alex D. , Plumb Jane , Mairs Robert J. , Gaze Mark N. , Workman Paul 
Published: February 2007

AbstractLaser scanning confocal microscopy has been used to follow the uptake and efflux of the 2‐fluoroglycoside of doxorubicin, ME2303, in live cultures of the human ovarian cancer cell line A2780 and its doxorubicin‐resistant variant A2780AD. Our methods combine confocal laser scanning microscopy and image analysis to examine the dynamics of anthracycline drugs in cancer cells. Cytotoxicity determined by MTT dye reduction showed that A2780AD cells were more than 400 times less sensitive to doxorubicin compared to A2780 cells but almost 9 times more sensitive to ME2303 compared to doxorubicin. The naturally fluorescent drug was tracked within live cells at 37°C to provide time‐course information in relation to nuclear and Golgi‐associated cellular domains, as indicated by BODIPY FL ceramide‐associated fluorescence. In both cell types, ME2303 was characterised by strong nuclear membrane and peri‐nucleolar fluorescence and as a localised punctate pattern within the nucleus. A2780AD cells accumulated ME2303 in their nuclei at a much reduced rate compared to the doxorubicin‐sensitive cells, and ME2303 efflux from resistant cell nuclei was approximately twice as fast as from A2780 cells. The relative uptake of ME2303 into Golgi‐associated domains and the nucleus were monitored simultaneously during the initial 35 min of exposure to 10 μM ME2303 and during the first 45 min of a “chase” culture following exposure to 20 μM ME2303. ME2303 was detectable within the Golgi‐associated domains of A2780 cells several minutes later and in less relative concentration than in A2780AD cells 15 min into a chase culture. Our results suggest the direct involvement of differences in drug processing via the Golgi apparatus in the expression of P‐glycoprotein‐related drug resistance.
Frequent deletion in chromosome 4 and duplication of chromosome 15 in liver epithelial cells derived from long‐term culture of C3H mouse hepatocytes
Authors: Nishimori Hiroyuki , Ogawa Katsuhiro , Tateno Hiroyuki 
Published: February 2007

AbstractProlonged culture of hepatocytes isolated from mouse liver results in the spontaneous development of colonies of liver epithelial cells that can proliferate indefinitely in vitro. We established 5 such cell lines from C3H/HeJ mice (C3H) and 22 cell lines from C3H/HeJ × C57BL/6J F1 mice (C3B6F1) to investigate whether any specific karyotype alterations may be associated with the development of such cells. These lines retained some properties of hepatocytes as well as showing bile‐duct‐cell characteristics, and comprised mainly near‐diploid and/or hypotetraploid cells. Karyotypic analysis of the C3H cell lines indicated that most cells have loss of chromosome 4 or deletion involving the C7 portion, while at least I (for near‐diploid cells) or 2 (for hypotetraploid cells) copies of chromosome 4 were usually intact. In addition, gain of an extra chromosome 15 was frequently observed in these cell lines. Analysis of the microsatellite DNA polymorphic markers in 22 C3B6F1 lines revealed that a majority of them showed loss of heterozygosity (LOH) for, at least, 1 of 3 polymorphic loci on chromosome 4, but not for 2 loci on chromosomes 7 and 11. Mouse chromosomes 4 and 15, therefore, may contain genes related to the ability of such liver epithelial cells to grow indefinitely in vitro [The locus on chromosome 4 was designated as liver‐cell immortalization (LCI) locus].