Angiogenic balance in human melanoma: Expression of VEGF, bFGF, IL-8, PDGF and angiostatin in relation to vascular density of xenograftsin vivo

Westphal, Johan R.; Hullenaar, Rianne Van't; Peek, Ron; Willems, Riki W.; Crickard, Kent; Crickard, Ulla; Askaa, Jon; Clemmensen, Inge; Ruiter, Dirk J.; Waal, R.M.W. de

doi:10.1002/(sici)1097-0215(20000615)86:6<768::aid-ijc3>3.0.co;2-e

Cited by 100 publications

(12 citation statements)

References 24 publications

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“…Although all cell lines gave rise to tumors within 1 week after inoculation, the growth characteristics of the various BLM transfectants appeared to be markedly different from those of the controls. In line with previous studies (11,16,17), the parental cell line and the vector controls grew to tumors exceeding a volume of 2.5 cm 3 within 4 weeks after tumor induction. In contrast, the RSV-CDT6 transfectants were about 6-fold smaller than the controls 4 weeks after tumor induction (Fig.…”

Section: Cdt6 Is a Secreted Protein Expressed In The Stromalsupporting

confidence: 89%

“…Ten g of RNA was size-fractionated on a 1% agarose, 2 M formaldehyde gel in MOPS buffer and blotted to Hybond N filter in 10ϫ SSC after transfer blots were hybridized with random prime-labeled cDNA probes. For reverse transcriptase-PCR analysis, 5 g of total RNA was reverse-transcribed in the presence of oligo(dT) [12][13][14][15][16][17][18] and 200 units of Superscript Range H reverse transcriptase (Invitrogen) according to the manufacturers' instructions. PCR was performed in a thermocycler (PerkinElmer Life Sciences) for 23 cycles.…”

Section: Methodsmentioning

confidence: 99%

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The Angiopoietin-like Factor Cornea-derived Transcript 6 Is a Putative Morphogen for Human Cornea

Peek¹,

Kammerer²,

Frank³

et al. 2002

Journal of Biological Chemistry

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The human cornea-specific protein cornea-derived transcript 6 (CDT6) is a member of the angiopoietin gene family. We report the structural and functional characterization of CDT6. We demonstrate that CDT6 is a secreted protein that folds into disulfide-linked homotetramers by coiled-coil interactions. The finding that CDT6 is expressed at high levels in the avascular corneal stromal layer suggested that the protein, similar to certain angiopoietins, acts as a negative regulator of angiogenesis. To test this hypothesis and to assay the effect of the protein on a growing tissue with high vascular density, CDT6 was expressed in a mouse xenograft model. Expression of CDT6 led to a reduction in tumor growth and aberrant blood vessel formation by inducing massive fibrosis. Interestingly, expression of CDT6 also resulted in the deposition of extracellular matrix components typical for the mature corneal stromal layer. These observations strongly suggest a role as morphogen for CDT6 in inducing a corneal phenotype in vivo.

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Section: Cdt6 Is a Secreted Protein Expressed In The Stromalsupporting

confidence: 89%

Section: Methodsmentioning

confidence: 99%

The Angiopoietin-like Factor Cornea-derived Transcript 6 Is a Putative Morphogen for Human Cornea

Peek¹,

Kammerer²,

Frank³

et al. 2002

Journal of Biological Chemistry

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“…Only when parental cells were transfected with VEGF 165 , a reduction in tumor volume was observed compared to VEGF 165 b-overexpressing tumors [23]. Similarly, significant differences in tumor volumes were only obtained in Mel57 (<600 pg/mL, [31]) and A375 (~600 pg/mL, [32]) melanoma xenograft models when VEGF 165 - and VEGF 165 b-overexpressing clones were compared [14,23]. On the contrary, in LS174t cells, which secrete high VEGF levels (~2000 pg/mL, [33]), overexpression of VEGF 165 b or VEGF 121 b [15], or administration of VEGF 165 b recombinant protein [24] led to tumor shrinkage compared to controls (parental cells with no further VEGF overexpression).…”

Section: Discussionmentioning

confidence: 99%

VEGF121b and VEGF165b are weakly angiogenic isoforms of VEGF-A

et al. 2010

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BackgroundDifferent isoforms of VEGF-A (mainly VEGF121, VEGF165 and VEGF189) have been shown to display particular angiogenic properties in the generation of a functional tumor vasculature. Recently, a novel class of VEGF-A isoforms, designated as VEGFxxxb, generated through alternative splicing, have been described. Previous studies have suggested that these isoforms may inhibit angiogenesis. In the present work we have produced recombinant VEGF121/165b proteins in the yeast Pichia pastoris and constructed vectors to overexpress these isoforms and assess their angiogenic potential.ResultsRecombinant VEGF121/165b proteins generated either in yeasts or mammalian cells activated VEGFR2 and its downstream effector ERK1/2, although to a lesser extent than VEGF165. Furthermore, treatment of endothelial cells with VEGF121/165b increased cell proliferation compared to untreated cells, although such stimulation was lower than that induced by VEGF165. Moreover, in vivo angiogenesis assays confirmed angiogenesis stimulation by VEGF121/165b isoforms. A549 and PC-3 cells overexpressing VEGF121b or VEGF165b (or carrying the PCDNA3.1 empty vector, as control) and xenotransplanted into nude mice showed increased tumor volume and angiogenesis compared to controls. To assess whether the VEGFxxxb isoforms are differentially expressed in tumors compared to healthy tissues, immunohistochemical analysis was conducted on a breast cancer tissue microarray. A significant increase (p < 0.05) in both VEGFxxxb and total VEGF-A protein expression in infiltrating ductal carcinomas compared to normal breasts was observed. A positive significant correlation (r = 0.404, p = 0.033) between VEGFxxxb and total VEGF-A was found.ConclusionsOur results demonstrate that VEGF121/165b are not anti-angiogenic, but weakly angiogenic isoforms of VEGF-A. In addition, VEGFxxxb isoforms are up-regulated in breast cancer in comparison with non malignant breast tissues. These results are to be taken into account when considering a possible use of VEGF121/165b-based therapies in patients.

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“…However, the cell lines used in these studies are less tumorigenic and have a lower metastatic potential than the highly metastatic human M14 melanoma cell line that we use here [15-17]. …”

Section: Bachgroundmentioning

confidence: 99%

Overexpression of the ATP binding cassette gene ABCA1 determines resistance to Curcumin in M14 melanoma cells

et al. 2009

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BackgroundCurcumin induces apoptosis in many cancer cells and it reduces xenograft growth and the formation of lung metastases in nude mice. Moreover, the plant derived polyphenol has been reported to be able to overcome drug resistance to classical chemotherapy. These features render the drug a promising candidate for tumor therapy especially for cancers known for their high rates concerning therapy resistance like melanoma.ResultsWe show here that the melanoma cell line M14 is resistant to Curcumin induced apoptosis, which correlates with the absence of any effect on NFκB signaling. We show that CXCL1 a chemokine that is down regulated in breast cancer cells by Curcumin in an NFκB dependant manner is expressed at variable levels in human melanomas. Yet in M14 cells, CXCL1 expression did not change upon Curcumin treatment. Following the hypothesis that Curcumin is rapidly removed from the resistant cells, we analyzed expression of known multi drug resistance genes and cellular transporters in M14 melanoma cells and in the Curcumin sensitive breast cancer cell line MDA-MB-231. ATP-binding cassette transporter ABCA1, a gene involved in the cellular lipid removal pathway is over-expressed in resistant M14 melanoma as compared to the sensitive MDA-MB-231 breast cancer cells. Gene silencing of ABCA1 by siRNA sensitizes M14 cells to the apoptotic effect of Curcumin most likely as a result of reduced basal levels of active NFκB. Moreover, ABCA1 silencing alone also induces apoptosis and reduces p65 expression.ConclusionResistance to Curcumin thus follows classical pathways and ABCA1 expression should be considered as response marker.

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Angiogenic balance in human melanoma: Expression of VEGF, bFGF, IL-8, PDGF and angiostatin in relation to vascular density of xenograftsin vivo

Cited by 100 publications

References 24 publications

The Angiopoietin-like Factor Cornea-derived Transcript 6 Is a Putative Morphogen for Human Cornea

The Angiopoietin-like Factor Cornea-derived Transcript 6 Is a Putative Morphogen for Human Cornea

VEGF121b and VEGF165b are weakly angiogenic isoforms of VEGF-A

Overexpression of the ATP binding cassette gene ABCA1 determines resistance to Curcumin in M14 melanoma cells

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