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See detailThe 6-Kilobase C-Erbb2 Promoter Contains Positive and Negative Regulatory Elements Functional in Human Mammary Cell Lines
Grooteclaes, Madeleine; Pasleau, Françoise ULg; Dijkmans, Huguette et al

in Cancer Research (1994), 54(15), 4193-9

A 6-kilobase fragment extending at the 5'-end of the c-erbB2 protooncogene was isolated from a normal human lymphocyte genomic DNA library. The full-length fragment and five subfragments with identical 3 ... [more ▼]

A 6-kilobase fragment extending at the 5'-end of the c-erbB2 protooncogene was isolated from a normal human lymphocyte genomic DNA library. The full-length fragment and five subfragments with identical 3'-ends were obtained by progressive unidirectional deletion from the 5'-end and were cloned in front of the luciferase reporter gene. The hybrid genes were analyzed for transcriptional activity in human mammary cell lines synthesizing low (HBL-100 and T-47D), moderate (MDA-MB-453), or high (BT-474) amounts of the c-erbB2 mRNA and were also analyzed in HeLa cells. Gene-specific expression was observed, indicating the presence of multiple cis-acting sequences in the c-erbB2 promoter. A major negative element is located in the -2- to -4-kilobase region. It is flanked on both sides by positive elements that display enhanced transcriptional activity in the BT-474 tumor cells only. While predominant in the low-expressing cells, the effect of the repressor appears to be overcome by the distal transactivator in the high-expressing BT-474 cells, resulting in a 15 to 50 times increase in luciferase activity relative to the HBL-100 and T-47D cells, respectively. Cell-specific expression relies on the trans-acting factors present in the different cell lines. The formation of cell-specific protein-DNA complexes was demonstrated by gel retardation assay. [less ▲]

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See detailExpression of Bone Sialoprotein, a Bone Matrix Protein, in Human Breast Cancer
Bellahcene, Akeila ULg; Merville, Marie-Paule ULg; Castronovo, Vincenzo ULg

in Cancer Research (1994), 54(11), 2823-6

Microcalcifications are often associated with human mammary lesions, particularly with breast carcinomas. To date, the molecular mechanism that leads to the deposition of hydroxyapatite in the mammary ... [more ▼]

Microcalcifications are often associated with human mammary lesions, particularly with breast carcinomas. To date, the molecular mechanism that leads to the deposition of hydroxyapatite in the mammary tissue has not been elucidated. Bone sialoprotein (BSP) is a glycoprotein the expression of which coincides with the appearance of the first hydroxyapatite crystals during bone development. In this study, we report the observation that BSP, a bone matrix protein, is expressed in human mammary cancer cells. Using an immunoperoxidase technique, we studied the expression of BSP in 79 breast lesions, including 28 benign and 51 malignant specimens. Two polyclonal antibodies, one directed against intact human BSP and the other against a synthetic peptide of BSP (residues 277-294), were used and gave identical results. Normal mammary glands expressed undetectable or barely detectable amounts of BSP, and the majority of the benign lesions examined were generally unstained (0) or weakly stained (1+). Most of the breast carcinoma specimens (around 87%) showed a significant increase (P = 0.0001) in BSP expression. Breast carcinomas with microcalcifications had the highest immunoreactivity (2+ or 3+) to BSP antibodies. This is the first demonstration that BSP expression is significantly increased in breast cancer. Expression of BSP by breast cancer cells could play a major role in the deposition of microcalcifications and in the preferred bone homing of breast cancer cells. [less ▲]

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See detailAgm-1470, a Potent Angiogenesis Inhibitor, Prevents the Entry of Normal but Not Transformed Endothelial Cells into the G1 Phase of the Cell Cycle
Antoine, Nadine ULg; Greimers, Roland ULg; De Roanne, C. et al

in Cancer Research (1994), 54(8), 2073-6

AGM-1470 is a potent angiogenesis inhibitor that is very effective in inhibiting endothelial cell proliferation in both in vitro and in vivo models and that prevents tumor growth in vivo. Although this ... [more ▼]

AGM-1470 is a potent angiogenesis inhibitor that is very effective in inhibiting endothelial cell proliferation in both in vitro and in vivo models and that prevents tumor growth in vivo. Although this molecule appears to be a most promising anticancer drug, its mechanism of action has not yet been elucidated. In this study, we examined the effects of AGM-1470 on the cell cycle of normal and transformed endothelial cells. We showed that AGM-1470, at picomolar concentrations, specifically inhibits the proliferation of both bovine aortic endothelial cells and human umbilical vein endothelial cells. AGM-1470 was ineffective in significantly inhibiting the proliferation of Ea.hy926 cells, a hybrid cell line obtained by the fusion of human umbilical vein endothelial cells with a human carcinoma cell line, or cEnd.1 cells, a polyoma middle T oncogene-transformed endothelioma cell line derived from mouse embryo. Using a double labeling technique with anti-Ki67 antibodies and propidium iodide, we demonstrated, with flow cytometry analysis, that AGM-1470 specifically prevents the entry of endothelial cells into the G1 phase of the cell cycle. We also showed that AGM-1470 was ineffective in inhibiting endothelial cell migration toward laminin or capillary-like tube formation inside a type I collagen matrix induced by phorbol esters. Our data strongly suggest that AGM-1470 is a molecule that specifically inhibits a cell cycle control pathway active in normal cells but which could be bypassed or altered in transformed cells. [less ▲]

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See detailTamoxifen restores the E-cadherin function in human breast cancer MCF-7/6 cells and suppresses their invasive phenotype
Bracke, M. E.; Charlier, Corinne ULg; Bruyneel, E. A. et al

in Cancer Research (1994), 54

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See detailTumor Cell Surface-Associated Binding Site for the M(R) 72,000 Type Iv Collagenase
Emonard, H. P.; Remacle, A. G.; Noël, Agnès ULg et al

in Cancer Research (1992), 52(20), 5845-8

We have studied the capacity of two human breast adenocarcinoma cells, MDA-MB231 and MCF-7, to bind exogenous M(r) 72,000 type IV collagenase by both morphological and radioreceptor binding assays. By ... [more ▼]

We have studied the capacity of two human breast adenocarcinoma cells, MDA-MB231 and MCF-7, to bind exogenous M(r) 72,000 type IV collagenase by both morphological and radioreceptor binding assays. By indirect immunofluorescence, staining with a specific anti-M(r) 72,000 type IV collagenase antibody was strongly induced when cells were preincubated with the purified enzyme. Scatchard plot analysis indicated the existence of a binding site for the M(r) 72,000 type IV collagenase with high affinity for both cell lines (Kd = 2 x 10(-9) M). These results are the first demonstration of the existence of a tumor cell membrane-associated putative receptor for a member of the matrix metalloproteinase family, as previously evidenced for the urokinase-type plasminogen activator. [less ▲]

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See detailExtracellular matrix receptors and mouse skin carcinogenesis: altered expression linked to appearance of early markers of tumor progression.
Tennenbaum, T.; Yuspa, S. H.; Grover, A. et al

in Cancer Research (1992), 52(10), 2966-76

Interaction of cells with the basement membrane is important for cell proliferation and differentiation. Disruption of the basement membrane is an early event during progression of benign tumors to cancer ... [more ▼]

Interaction of cells with the basement membrane is important for cell proliferation and differentiation. Disruption of the basement membrane is an early event during progression of benign tumors to cancer. Using the techniques of immunohistochemistry and immunofluorescence, we show that cell-matrix interactions via the cell surface integrin receptors alpha 3 beta 1, alpha 5 beta 1, alpha 6 beta 4, the Mr 67,000 laminin receptor (67LR) laminin-binding protein, and the secreted matrix protein laminin are strictly regulated during differentiation of mouse epidermis. While alpha 6 beta 4 and alpha 5 beta 1 are polarized to the basal surface of basal cells in contact with the basement membrane, alpha 3 beta 1 and the non-integrin 67LR are primarily detected in the cell periphery of suprabasal cells, where cell to cell contacts are found. Sequential changes in expression of matrix receptors occur following multistage carcinogenesis of mouse skin. In an analysis of benign and malignant skin tumors induced by chemical carcinogens or oncogene transduction, we found that alpha 3 beta 1 and alpha 5 beta 1 as well as the non-integrin 67LR are sequentially down-regulated in the progression from benign to malignant, while alpha 6 beta 4 is the predominant receptor expressed in the carcinomas. Tumor expression of alpha 6 beta 4 is not polarized and is dissociated from its colocalized normal partner bullous pemphigoid antigen, which remains restricted to the basement membrane. The changes in matrix receptors are linked to appearance of keratin 13 in suprabasal regions, but always in alpha 6 beta 4 negative cells. The predominance of alpha 6 beta 4 in the proliferating cells during progression is associated with decreased expression of keratin 13 in carcinomas. These results suggest that matrix interactions with its receptors are important determinants of ordered differentiation in normal skin and show characteristic alterations during carcinogenesis that parallel changes in differentiation of the tumors. [less ▲]

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See detailEnhancement of 1,3-bis(2-chloroethyl)-1-nitrosourea resistance by gy gamma-irradiation or drug pretreatment in rat hepatoma cells
Habraken, Yvette ULg; Laval, Françoise

in Cancer Research (1991), 51(4), 499-503

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See detailInvasion of Reconstituted Basement Membrane Matrix Is Not Correlated to the Malignant Metastatic Cell Phenotype
Noël, Agnès ULg; Calle, A.; Emonard, H. et al

in Cancer Research (1991), 51(1), 405-14

Interactions between tumor cells and basement membranes represent a critical step in the progression of neoplasia and in the metastatic process. Reconstituted basement membrane matrix, matrigel, has been ... [more ▼]

Interactions between tumor cells and basement membranes represent a critical step in the progression of neoplasia and in the metastatic process. Reconstituted basement membrane matrix, matrigel, has been recently used with the aim of developing an in vitro assay of tumor cell invasiveness. We have extended these studies by comparing the invasiveness of a large series of normal and malignant epithelial and mesenchymal cells of human and animal origin cultured on matrigel. Normal cells (fibroblasts, glomerular mesangial cells, keratinocytes), human fibrosarcoma cells (HT1080), and reticular sarcoma cells (M5076) clearly established invasive capabilities in the matrix. However, all the other tested cell lines, malignant or virally transformed cells invasive in vivo (MCF7, T47D, SA52, SW613, MO4, A431, BeWo), as well as normal nontransformed cells (MOH22) were incapable of penetration. The morphological features of matrigel invasion by normal fibroblasts and HT1080 cells are described at the light and electron microscope levels. The extent of degradation of a radiolabeled matrigel is minimal and similar in several cell lines reported to be noninvasive or invasive in vivo. Our data suggest that matrigel does not provide a universal model to correlate the invasiveness of cells in vivo and in vitro. [less ▲]

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See detailLaminin receptor complementary DNA-deduced synthetic peptide inhibits cancer cell attachment to endothelium.
Castronovo, Vincenzo ULg; Taraboletti, G.; Sobel, M. E.

in Cancer Research (1991), 51(20), 5672-8

Stable attachment of cancer cells to the endothelium is a key step in the formation of metastasis. In this study, we have investigated the possibility that interaction between laminin and its Mr 67,000 ... [more ▼]

Stable attachment of cancer cells to the endothelium is a key step in the formation of metastasis. In this study, we have investigated the possibility that interaction between laminin and its Mr 67,000 high-affinity receptor (67 LR) could play a major role in this process. Scatchard analysis of laminin-binding studies showed that bovine aortic endothelial cells exhibit 46,000 high-affinity receptors that mediate, at least in part, the attachment of highly invasive melanoma cells. This endothelial cell-melanoma cell interaction was significantly inhibited by soluble laminin and by anti-laminin antibodies. Peptide G, an eicosapeptide derived from the complementary DNA sequence of the 67 LR precursor (IPCNNKGAHSVGLMWWMLAR) that specifically binds to laminin and presumably contains the active ligand-binding site of the receptor, specifically prevented attachment of the melanoma cells to both the bovine aortic endothelial cell monolayer and human umbilical vein endothelium. Thus, peptide G may selectively interfere with the metastatic cascade by inhibiting tumor cell attachment to endothelium via the laminin-67 LR pathway and is a potential new antimetastatic agent. [less ▲]

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See detailRelease of 7-alkylguanines from N-(2-chloroethyl)-N'-cyclohexyl-N-nitrosourea-modified DNA by 3 methyladenine DNA glycosylase
Habraken, Yvette ULg; Carter, C.; Kirk, M. et al

in Cancer Research (1991), 51

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See detailImmunofluorescence localization of fibronectin in chondrosarcoma cartilage matrix.
Kimata, K.; Foidart, Jean-Michel ULg; Pennypacker, J. P. et al

in Cancer Research (1982), 42(6), 2384-91

In this study, we have compared the extracellular matrix components and the in vitro adhesion characteristics of normal rat epiphysial chondrocytes with those from the Swarm rat chondrosarcoma, which has ... [more ▼]

In this study, we have compared the extracellular matrix components and the in vitro adhesion characteristics of normal rat epiphysial chondrocytes with those from the Swarm rat chondrosarcoma, which has many of the biochemical characteristics of normal cartilage. With the use of immunofluorescence techniques, tissue slices and chondrocytes in culture were tested for the presence of collagen types I and II, cartilage-characteristic proteoglycan, and fibronectin. Both normal and tumor matrix contained type II collagen and cartilage proteoglycan, but only the tumor matrix contained fibronectin. In culture, tumor-derived chondrocytes continued to accumulate fibronectin in their matrix, even after deposition of type II collagen and proteoglycans, while normal chondrocytes did not. When the attachment characteristics of both types of chondrocytes were compared, tumor chondrocytes required fibronectin for attachment, while normal chondrocytes used another attachment factor that had been identified previously as chondronectin. These studies suggest that, although biochemically similar to normal chondrocytes, tumor chondrocytes are no longer able to express the regulatory mechanisms for fibronectin accumulation. [less ▲]

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See detailReversal by glucocorticoid hormones of the loss of a fibronectin and probollagen matrix around transformed human cells.
Furcht, L. T.; Mosher, D. F.; Wendelschafer-Crabb, G. et al

in Cancer Research (1979), 39(6Pt 1), 2077-83

Confluent cultured human skin fibroblasts had an extracellular fibrillar matrix of fibronectin and procollagen. Human skin fibroblasts transformed by SV40 did not have such a matrix. Treatment of ... [more ▼]

Confluent cultured human skin fibroblasts had an extracellular fibrillar matrix of fibronectin and procollagen. Human skin fibroblasts transformed by SV40 did not have such a matrix. Treatment of transformed fibroblasts with 10(-5) to 10(-8) M dexamethasone and 10(-5) to 10(-7) M cortisol, but not testosterone or progesterone, caused partial restoration of the matrix. Glucocorticoid-treated transformed human fibroblasts can serve as a model for partial reversion toward normal or differentiation of transformed human fibroblasts. [less ▲]

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See detailHumoral antibody response to bovine leukemia virus infection in cattle and sheep.
Bex, Françoise; Bruck, Claudine; Mammerickx, Marc et al

in Cancer Research (1979), 39(3), 1118-1123

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