Bcl‐2 overexpression in human melanoma cells increases angiogenesis through VEGF mRNA stabilization and HIF‐1mediated transcriptional activity

Iervolino, Angela; Trisciuoglio, Daniela; Ribatti, Doménico; Candiloro, Antonio; Biroccio, Annamaria; Zupi, Gabriella; Bufalo, Donatella Del

doi:10.1096/fj.02-0122fje

Cited by 117 publications

(151 citation statements)

References 47 publications

(78 reference statements)

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“…We further investigated the effect of PI3K and MAPK inhibitors on VEGF mRNA-induction by bcl-2/hypoxia in MB5 and MB6 bcl-2-overexpressing clones ( Figure 3A). As expected (Iervolino et al, 2002), 24-h hypoxia induced about a three-to fourfold increase in mRNA expression in MB5 and MB6 clones compared with normoxic conditions. The two inhibitors reduced the induction of VEGF mRNA expression by bcl-2 in both bcl-2 transfectants.…”

supporting

confidence: 86%

“…EMSA was performed, as described previously (Iervolino et al, 2002), after exposure to hypoxia for 24 h in the presence or absence of UO126, PD98059, Wort, or LY29402. The following double-strand oligomer as radiolabeled probes or cold competitor: HIF-1 (Ϫ985 to -951 of human VEGF 5Ј gene promoter), 5ЈTCGACCACAGTGCATACGTGGGCTCCAACAGGTCCTCTTC-3Ј.…”

Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

“…Receptor tyrosine kinase activity also leads to signaling via the ERK and p38 MAP kinase pathways. PI3K and AKT also have been implicated in the induction of HIF-1 expression by hypoxia (Berra et al, 2000;Semenza, 2002b).We previously demonstrated that bcl-2 overexpression in human melanoma cells exposed to hypoxic conditions increases VEGF promoter activity and mRNA stability and enhances HIF-1 DNA binding activity (Iervolino et al, 2002 However, the steps that participate in VEGF expression mediated by HIF-1 activity in response to bcl-2 overexpression in human melanoma cells exposed to hypoxia are still unknown. We have demonstrated that the MAPK pathway is implicated in signal transduction by bcl-2 (Trisciuoglio et al, 2004).…”

mentioning

confidence: 99%

“…On the contrary, no modulation of ERK1/2 phosphorylation was evidenced in the MN8 control clone after down-regulation of the bcl-2 protein ( Figure 4C). As mentioned above and previously described, bcl-2 is able to increase VEGF expression and transcriptional activity (Iervolino et al, 2002). To exclude the possibility that the decrease in cell survival induced by bcl-2 RNAi (Figure 4) affects VEGF expression, conditions were chosen under which bcl-2 RNAi did not activate the apoptotic program in the cells.…”

mentioning

confidence: 99%

“…Treatment with either UO126 or Wort strongly reduced the DNA binding activity of bcl-2 transfectants, clearly suggesting that HIF-1 DNA binding activity was MAPK and PI3K dependent. On the contrary, no significant reduction of DNA binding activity was observed in the parental cells exposed to either inhibitor.Because we previously found the ability of bcl-2 to increase hypoxic responsive element (HRE) promoter activity in hypoxia (Iervolino et al, 2002), we evaluated the role of the PI3K and MAPK signaling pathways in the transcriptional activation of HIF-1 by bcl-2/hypoxia. Parental and bcl-2-overexpressing cells were transiently transfected with a pHRE-Luc reporter plasmid, which contains four concatamerized HIF-1 binding sites, and then exposed to normoxia or hypoxia for 24 h in the presence or absence of PI3K and MAPK inhibitors.…”

mentioning

confidence: 99%

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Involvement of PI3K and MAPK Signaling in bcl-2-induced Vascular Endothelial Growth Factor Expression in Melanoma Cells

Trisciuoglio¹,

Iervolino²,

Zupi³

et al. 2005

MBoC

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We have previously demonstrated that bcl-2 overexpression in tumor cells exposed to hypoxia increases the expression of vascular endothelial growth factor (VEGF) gene through the hypoxia-inducible factor-1 (HIF-1). In this article, we demonstrate that exposure of bcl-2 overexpressing melanoma cells to hypoxia induced phosphorylation of AKT and extracellular signal-regulated kinase (ERK)1/2 proteins. On the contrary, no modulation of these pathways by bcl-2 was observed under normoxic conditions. When HIF-1␣ expression was reduced by RNA interference, AKT and ERK1/2 phosphorylation were still induced by bcl-2. Pharmacological inhibition of mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) signaling pathways reduced the induction of VEGF and HIF-1 in response to bcl-2 overexpression in hypoxia. No differences were observed between control and bcl-2-overexpressing cells in normoxia, in terms of VEGF protein secretion and in response to PI3K and MAPK inhibitors. We also demonstrated that RNA interference-mediated down-regulation of bcl-2 expression resulted in a decrease in the ERK1/2 phosphorylation and VEGF secretion only in bcl-2-overexpressing cell exposed to hypoxia but not in control cells. In conclusion, our results indicate, for the first time, that bcl-2 synergizes with hypoxia to promote expression of angiogenesis factors in melanoma cells through both PI3K-and MAPK-dependent pathways. INTRODUCTIONAngiogenesis is a complex and multilevel process of new capillary formation on the basis of already existing blood vessels. Physiologically, it is a very strictly regulated process, which results in a balance between stimulatory (angiogenic) and inhibitory (angiostatic) factors to control the correct development of blood vessels. Angiogenesis is a rate-limiting step in tumor growth and progression, and many tumors develop a severely hypoxic microenvironment and secrete angiogenesis-stimulating factors (Folkman et al., 1971;Talks et al., 2000). Vascular endothelial growth factor (VEGF), a potent and specific mitogen for vascular endothelial cells, is a critical mediator of angiogenesis. Its expression is induced in cancer cells as a result of hypoxia and multiple genetic alterations, including p53 and PTEN loss-of-function, RAS and SRC gain-of-function, and autocrine tyrosine kinase signaling pathways (Mukhopadhyay et al., 1995;Arbiser et al., 1997;Petit et al., 1997;Akagi et al., 1998;Ellis et al., 1998;Yen et al., 2000). Based on the analysis of melanoma and breast cancer cells recently reported by our group Iervolino et al., 2002;Del Bufalo et al., 2003;Trisciuoglio et al., 2004), this list can now be extended to include the increased VEGF expression resulting from bcl-2 overexpression. Hypoxia-and growth factor-induced VEGF expression in tumor cells is regulated by the hypoxia inducible factor 1 (HIF-1) (Jiang et al., 1997;Mazure et al., 1997;Ravi et al., 2000;Zhong et al., 2000;Zundel et al., 2000;Laughner et al., 2001;Fukuda et al., 2002), a transcription factor that is compose...

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supporting

confidence: 86%

Section: Electrophoretic Mobility Shift Assay (Emsa)mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Involvement of PI3K and MAPK Signaling in bcl-2-induced Vascular Endothelial Growth Factor Expression in Melanoma Cells

Trisciuoglio¹,

Iervolino²,

Zupi³

et al. 2005

MBoC

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Hypoxia‐inducible factor‐1α in non small cell lung cancer: Relation to growth factor, protease and apoptosis pathways

Swinson

Jones

Cox

et al. 2004

Intl Journal of Cancer

151

107

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Hypoxia-inducible factor (HIF)-1␣ is the regulatory subunit of HIF-1 that is stabilized under hypoxic conditions. Under different circumstances, HIF-1␣ may promote both tumorigenesis and apoptosis. There is conflicting data on the importance of HIF-1␣ as a prognostic factor. This study evaluated HIF-1␣ expression in 172 consecutive patients with stage I-IIIA non small cell lung cancer (NSCLC) using standard immunohistochemical techniques. The extent of HIF-1␣ nuclear immunostaining was determined using light microscopy and the results were analyzed using the median (5%) as a low cut-point and 60% as a high positive cut-point.

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Bcl‐2 overexpression in melanoma cells increases tumor progression‐associated properties and in vivo tumor growth

Trisciuoglio¹,

Desideri²,

Ciuffreda³

et al. 2005

Journal Cellular Physiology

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In this study, we demonstrated that bcl-2 overexpression in human melanoma cells consistently enhanced the activity of multiple metastasis-related proteinases, in vitro cell invasion, and in vivo tumor growth. In particular, by using the M14 parental cell line, the MN8 control clone, and two bcl-2 overexpressing derivatives, we found that bcl-2 overexpressing cells exposed to hypoxia, when compared to parental cells, expressed higher level of several metalloproteases (MMPs) such as MMP-2, MMP-7, MT1-MMP, and tissue inhibitors of metalloproteases-1 and -2. Moreover, bcl-2 overexpression in melanoma cells enhanced in vitro invasion on matrigel and, in vivo tumor growth. The more aggressive behavior of bcl-2 transfectants tumors is significantly associated to an increase in MMP-2 expression as well as in a more elevated microvessel density as compared to the parental line. Taken together, our data suggest that bcl-2 plays a pivotal role in the regulation of molecules associated with the migratory and invasive phenotype, contributing, in cooperation to hypoxia, to tumor progression.

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Bcl‐2 overexpression in human melanoma cells increases angiogenesis through VEGF mRNA stabilization and HIF‐1mediated transcriptional activity

Cited by 117 publications

References 47 publications

Involvement of PI3K and MAPK Signaling in bcl-2-induced Vascular Endothelial Growth Factor Expression in Melanoma Cells

Involvement of PI3K and MAPK Signaling in bcl-2-induced Vascular Endothelial Growth Factor Expression in Melanoma Cells

Hypoxia‐inducible factor‐1α in non small cell lung cancer: Relation to growth factor, protease and apoptosis pathways

Bcl‐2 overexpression in melanoma cells increases tumor progression‐associated properties and in vivo tumor growth

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