Transfection of human endothelial cells with plasminogen activator inhibitor type-1 promoter/reporter gene fragments reveals phorbol ester induction of gene expression

Descheemaeker, Koen; Nelles, Luc; Moreau, H; Strandberg, Leif; Ny, Tor; Collen, D.

doi:10.1016/0268-9499(92)90079-w

Cited by 14 publications

(16 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…DNA binding and dimerization are mediated by a basically charged domain preceding a helix ± span ± helix motif located in the C-terminus (Williams and Tjian, 1991a, b). Functional AP-2 binding sites have been identi®ed in the enhancer regions of viral genes such as simian virus 40 (SV40) (Mitchell et al, 1987), human T-cell leukemia virus type I (Nyborg and Dynan, 1990), and cellular genes including melanoma cellular adhesion molecule (MCAM/MUC18) (Jean et al, 1998a), tyrosine kinase receptor c-KIT (Huang et al, 1998), thrombin receptor (Schmidt et al, 1996), p21/WAF1 (Ropponen et al, 1999;Zeng et al, 1997), c-myc (Gaubatz et al, 1995), type IV collagenase/ gelatinase (Huhtala et al, 1990), intercellular adhesion molecule (ICAM-1) (Grether-Beck et al, 1996), inositol 1,4,5-triphosphate receptor (IP 3 R) (Williams and Tjian, 1991b), murine major histocompatibility complex (H-2K b ) (Kanno et al, 1989), human metallothionein-IIa (huMTIIa) (Haslinger and Karin, 1985;Lee et al, 1987), human proenkephalin (Hyman et al, 1989), human keratin K14 (Leask et al, 1991), keratin 10 ( Maytin et al, 1999), c-erb-2 (Bosher et al, 1995), plasminogen activator inhibitor type I (PAI-1) (Descheemaeker et al, 1992), and insulin-like growth factor binding protein-5 (Duan and Clemmons, 1995).…”

Section: Introductionmentioning

confidence: 99%

Dominant-negative transcription factor AP-2 augments SB-2 melanoma tumor growth in vivo

et al. 2001

View full text Add to dashboard Cite

We have previously demonstrated that the transition of melanoma to the metastatic phenotype is associated with a loss of expression of the transcription factor AP-2. To further investigate the role of AP-2 in the progression of human melanoma, we attempted to inactivate AP-2 in primary cutaneous SB-2 melanoma cells by using a dominant-negative AP-2, or AP-2B, gene. AP-2B is an alternatively spliced AP-2 variant capable of inhibiting AP-2 trans-activator function. Stable transfection of primary cutaneous melanoma SB-2 cells with the dominant-negative AP-2B gene was con®rmed by RT ± PCR and Northern blot analyses. Electromobility shift assay using nuclear extracts from these cell lines demonstrated decreased functional binding of AP-2B-transfected cells to the AP-2 consensus binding sequence compared with neo-transfected controls. In addition, CAT activity driven by a construct containing the AP-2 consensus binding sequence was downregulated in the AP-2B transfected cells, indicating AP-2 activity was quenched in the transfected cells. Orthotopic (subcutaneous) injection of the dominant-negative (AP-2B)-transfected cell lines into nude mice increased their tumorigenicity compared to control neo-transfected cells. The AP-2B-transfected cells displayed an increase in MMP-2 expression (by Northern blot) and MMP-2 activity (by zymography), which resulted in an increase in invasiveness through Matrigel-coated ®lters. The AP-2B-transfected tumors also displayed an increase in MMP-2 expression, microvessel density, and angiogenesis in vivo. These results demonstrate that inactivation of AP-2 contributes to the progression of melanoma, at least partially via deregulation of the MMP-2 gene.

show abstract

Section: Introductionmentioning

confidence: 99%

Dominant-negative transcription factor AP-2 augments SB-2 melanoma tumor growth in vivo

et al. 2001

View full text Add to dashboard Cite

show abstract

“…13) and HeLa nuclear extracts had shown a major shifted band corresponding to several protein/DNA complexes (17). In order to identify the proteins involved in these complexes, EMSA conditions have now been modified allowing resolution of four distinct complexes (C1, C2, and the C3a/C3b doublet in Fig.…”

Section: Hltf Sp1 and Sp3mentioning

confidence: 99%

“…Some cis-elements involved in gene regulations have been identified in the human PAI-1 promoter, such as those involved in the induction by transforming growth factor-␤ (9, 10), glucocorticoids (11,12), phorbol ester (13,14), p53 (15), and glucose (16).…”

mentioning

confidence: 99%

“…The same protein was independently isolated by other groups because it interacted with other DNA targets (the human immunodeficiency virus long terminal repeat and the simian virus enhancer, the myosin light chain locus enhancer, the rabbit uteroglobin promoter, a tumor necrosis factor response element), and shown to display DNA-dependent ATPase activity (22)(23)(24)(25). HLTF activates the PAI-1 promoter via specific interaction with the B box that was initially identified as a phorbol ester-responsive element (13), but was later shown only to be involved in basal expression (17). The B box is highly similar to the GT box (also called CACCC motif), which has been shown to bind Sp1 as well as other recently identified members of the expanding Sp family of transcription factors (26 -28).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Functional Interactions between Sp1 or Sp3 and the Helicase-like Transcription Factor Mediate Basal Expression from the Human Plasminogen Activator Inhibitor-1 Gene

Ding¹,

Benotmane²,

Suske³

et al. 1999

Journal of Biological Chemistry

View full text Add to dashboard Cite

Basal expression of the human plasminogen activator inhibitor-1 (PAI-1) is mediated by a promoter element named B box that binds the helicase-like transcription factor (HLTF), homologous to SNF/SWI proteins. Electrophoretic mobility shift assays performed on a set of B box point mutants demonstrated two HLTF sites flanking and partially overlapping with a GT box binding Sp1 and Sp3. Mutations affecting either the Sp1/Sp3 or the two HLTF sites inhibited by 6-and 2.5-fold, respectively, transient expression in HeLa cells of a reporter gene fused to the PAI-1 promoter. In Sp1/Sp3-devoid insect cells, co-expression of PAI-1-lacZ with Sp1 or Sp3 led to a 14 -26-fold induction while HLTF had no effect. Simultaneous presence of Sp1 or Sp3 and the short HLTF form (initiating at Met-123) provided an additional 2-3-fold synergistic activation suppressed by mutations that prevented HLTF binding. Moreover, a DNA-independent interaction between HLTFMet123 and Sp1/Sp3 was demonstrated by co-immunoprecipitation from HeLa cell extracts and glutathione S-transferase pull-down experiments. The interaction domains were mapped to the carboxyl-terminal region of each protein; deletion of the last 85 amino acids of HLTFMet123 abolished the synergy with Sp1. This is the first demonstration of a functional interaction between proteins of the Sp1 and SNF/ SWI families.

show abstract

“…In addition, inactivation of AP-2 in primary cutaneous melanoma cells (AP-2 positive) by means of a dominant-negative AP-2 gene (AP-2B) led to deregulation of the metalloproteinase MMP-2 gene and increased the cells tumor and metastatic potential in nude mice (Gershenwald et al, 2001). Furthermore, functional AP-2-binding elements have been identified in other genes involved in the progression of human melanoma such as p21/WAF1 (Zeng et al, 1997), intercellular adhesion molecule (I-CAM) (Grether-Beck et al, 1996), c-erbB-2/HER-2/neu (Hollywood and Hurst, 1993;Bosher et al, 1995;Turner et al, 1998), plasminogen activator inhibitor type I (PAI-1) (Descheemaeker et al, 1992), insulin-like growth factor binding protein-5 (Duan and Clemmons, 1995), transforming growth factor-alpha (TGF-a) (Wang et al, 1997), vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) (Gille et al, 1997(Gille et al, , 2000, E-cadherin (Batsche et al, 1998), hepatocyte growth factor (HGF) , and c-Myc (Gaubatz et al, 1995). Loss of AP-2 was also observed in clinical specimens of advanced primary and metastatic melanoma, which was associated with malignant transformation and elevated risk of tumor progression (Karjalainen et al, 1998;Baldi et al, 2001), thus supporting the hypothesis that loss of AP-2 is a crucial event in the progression and metastasis of melanoma.…”

mentioning

confidence: 99%

Role and regulation of the thrombin receptor (PAR-1) in human melanoma

2003

View full text Add to dashboard Cite

To determine treatment strategies and predict the clinical outcome of patients with melanoma it is important to understand the etiology of this disease. Recently, there has been some insight into molecular basis of melanoma including identification of a few of the regulatory factors and genes involved in this disease. For instance, the transcription factor AP-2 plays a tumor suppressor-like role in melanoma progression by regulating genes involved in tumor growth and metastasis. Previously, we have shown that the progression of human melanoma to the metastatic phenotype is associated with loss of AP-2 expression and deregulation of target genes such as MUC18/MCAM, c-KIT, and MMP-2. Increasing evidence demonstrates that the thrombin receptor (proteaseactivated receptor-1, PAR-1) plays a major role in tumor invasion and contributes to the metastatic phenotype of human melanoma. This review focuses on the role of the thrombin receptor in melanoma and its regulation by AP-2. We show that loss of AP-2 expression in metastatic melanoma cells correlates with overexpression of the thrombin receptor. Our analysis of AP-2/Sp1 complexes within the regulatory region of the thrombin receptor demonstrates that AP-2 binds the proximal 3 0 region of the promoter and diminishes PAR-1 expression. Levels of AP-2 and Sp1 proteins in a panel of melanoma cell lines demonstrated a marked decrease in the ratio of AP-2/ Sp1, a decrease that correlated with overexpression of PAR-1 in metastatic melanoma cells. We propose that loss of AP-2 results in increased expression of the thrombin receptor, which subsequently contributes to the metastatic phenotype of melanoma by upregulating the expression of adhesion molecules, proteases, and angiogenic molecules.

show abstract

Transfection of human endothelial cells with plasminogen activator inhibitor type-1 promoter/reporter gene fragments reveals phorbol ester induction of gene expression

Cited by 14 publications

References 38 publications

Dominant-negative transcription factor AP-2 augments SB-2 melanoma tumor growth in vivo

Dominant-negative transcription factor AP-2 augments SB-2 melanoma tumor growth in vivo

Functional Interactions between Sp1 or Sp3 and the Helicase-like Transcription Factor Mediate Basal Expression from the Human Plasminogen Activator Inhibitor-1 Gene

Role and regulation of the thrombin receptor (PAR-1) in human melanoma

Contact Info

Product

Resources

About