Aims: DMXAA (AS1404), a small-molecule vascular disrupting agent that has now completed Phase II clinical trial, induces endothelial cell apoptosis, increased vascular permeability and decreased tumour blood flow in vivo. Its action is incompletely understood and we wished to develop an in vitro system to study its effects.
Methods: Human tumour cell lines developed from aggressive tumours were grown on Matrigel to simulate a tumour microenvironment. Cells were analysed by light microscopy and by gene expression profiling.
Results: Several cell lines formed networks when grown on Matrigel and the NZM7 melanoma cell line was chosen for further study. Addition of DMXAA at a clinically achievable concentration (30 μg/mL) prevented network formation, but co-addition of SB203580 (10 μM), a selective inhibitor of p38 MAP kinase, reversed the effect of DMXAA and restored network formation. Analysis of expression genes for endothelial and related functions showed that cells growing on Matrigel expressed a pattern similar to that of NZM7 cells growing as xenografts in vivo but different from that of cells grown on standard tissue culture plates. Addition of DMXAA resulted in the inhibition of expression of several genes including the transcriptional activator Ets1 and matrix metalloproteinase-2 (MMP2), but co-addition of SB203580 did not reverse these effects of DMXAA on gene expression.
Conclusion: The results suggest that p38 MAP kinase plays an important role in the action of DMXAA and that growth of tumour cells on Matrigel provides a promising model for further studies on the action of this drug.
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This research was supported by the Auckland Cancer Society and by an Antisoma Postdoctoral Fellowship.
Tozer GM, Kanthou C, Baguley BC (2005) Disrupting tumour blood vessels. Nature Rev Cancer 5:423–435CrossRefGoogle Scholar
Rewcastle GW, Atwell GJ, Li ZA, Baguley BC, Denny WA (1991) Potential antitumor agents. 61. Structure-activity relationships for in vivo colon 38 activity among disubstituted 9-oxo-9H-xanthene-4-acetic acids. J Med Chem 34:217–222PubMedCrossRefGoogle Scholar
Galbraith SM, Rustin GJ, Lodge MA, Taylor NJ, Stirling JJ, Jameson M, Thompson P, Hough D, Gumbrell L, Padhani AR (2002) Effects of 5,6-dimethylxanthenone-4-acetic acid on human tumor microcirculation assessed by dynamic contrast-enhanced magnetic resonance imaging. J Clin Oncol 20:3826–3840PubMedCrossRefGoogle Scholar
Ching LM, Cao Z, Kieda C, Zwain S, Jameson MB, Baguley BC (2002) Induction of endothelial cell apoptosis by the antivascular agent 5,6-dimethylxanthenone-4-acetic acid. Br J Cancer 86:1937–1942PubMedCrossRefGoogle Scholar
Kestell P, Zhao L, Jameson MB, Stratford MR, Folkes LK, Baguley BC (2001) Measurement of plasma 5-hydroxyindoleacetic acid as a possible clinical surrogate marker for the action of antivascular agents. Clin Chim Acta 314:159–166PubMedCrossRefGoogle Scholar
Jameson MB, Thompson PI, Baguley BC, Evans BD, Harvey VJ, Porter DJ, McCrystal MR, Small M, Bellenger K, Gumbrell L, Halbert GW, Kestell P (2003) Clinical aspects of a phase I trial of 5,6-dimethylxanthenone-4-acetic acid (DMXAA), a novel antivascular agent. Br J Cancer 88:1844–1850PubMedCrossRefGoogle Scholar
McKeage MJ, Fong P, Jeffery M, Baguley BC, Kestell P, Ravic M, Jameson MB (2006) 5,6-Dimethylxanthenone-4-acetic acid in the treatment of refractory tumors: a phase I safety study of a vascular disrupting agent. Clin Cancer Res 12:1776–1784PubMedCrossRefGoogle Scholar
Rustin GJ, Bradley C, Galbraith S, Stratford M, Loadman P, Waller S, Bellenger K, Gumbrell L, Folkes L, Halbert G (2003) 5,6-dimethylxanthenone-4-acetic acid (DMXAA), a novel antivascular agent: phase I clinical and pharmacokinetic study. Br J Cancer 88:1160–1167PubMedCrossRefGoogle Scholar
McKeage MJ, AS1404-201 Study Group Investigators (2006) Phase Ib/II study of DMXAA combined with carboplatin and paclitaxel in non-small cell lung cancer (NSCLC). J Clin Oncol 24:7102Google Scholar
Parmar J, Marshall ES, Charters GA, Holdaway KM, Shelling AN, Baguley BC (2000) Radiation-induced cell cycle delays and p53 status of early passage melanoma cell lines. Oncol Res 12:149–155PubMedGoogle Scholar
Marshall ES, Holdaway KM, Shaw JH, Finlay GJ, Matthews JH, Baguley BC (1993) Anticancer drug sensitivity profiles of new and established melanoma cell lines. Oncol Res 5:301–309PubMedGoogle Scholar
Drinkwater SL, Smith A, Sawyer BM, Burnand KG (2002) Effect of venous ulcer exudates on angiogenesis in vitro. Br J Surg 89:709–713PubMedCrossRefGoogle Scholar
Sengupta S, Sellers LA, Matheson HB, Fan TP (2003) Thymidine phosphorylase induces angiogenesis in vivo and in vitro: an evaluation of possible mechanisms. Br J Pharmacol 139:219–231PubMedCrossRefGoogle Scholar
Rybak SM, Sanovich E, Hollingshead MG, Borgel SD, Newton DL, Melillo G, Kong D, Kaur G, Sausville EA (2003) “Vasocrine” formation of tumor cell-lined vascular spaces: implications for rational design of antiangiogenic therapies. Cancer Res 63:2812–2819PubMedGoogle Scholar
Ching LM, Zwain S, Baguley BC (2004) Relationship between tumour endothelial cell apoptosis and tumour blood flow shutdown following treatment with the antivascular agent DMXAA in mice. Br J Cancer 90:906–910PubMedCrossRefGoogle Scholar
Zhao L, Ching LM, Kestell P, Kelland LR, Baguley BC (2005) Mechanisms of tumor vascular shut-down induced by 5,6-dimethylxanthenone-4-acetic acid (DMXAA); increased tumor vascular permeability. Int J Cancer 116:322–326PubMedCrossRefGoogle Scholar
Ferrero E, Zocchi MR, Magni E, Panzeri MC, Curnis F, Rugarli C, Ferrero ME, Corti A (2001) Roles of tumor necrosis factor p55 and p75 receptors in TNF-alpha-induced vascular permeability. Am J Physiol - Cell Physiol 281:C1173–C1179PubMedGoogle Scholar
McMullen ME, Bryant PW, Glembotski CC, Vincent PA, Pumiglia KM (2005) Activation of p38 has opposing effects on the proliferation and migration of endothelial cells. J Biol Chem 280:20995–21003PubMedCrossRefGoogle Scholar
Hess AR, Seftor EA, Seftor RE, Hendrix MJ (2003) Phosphoinositide 3-kinase regulates membrane Type 1-matrix metalloproteinase (MMP) and MMP-2 activity during melanoma cell vasculogenic mimicry. Cancer Res 63:4757–4762PubMedGoogle Scholar
Hendrix MJ, Seftor EA, Hess AR, Seftor RE (2003) Vasculogenic mimicry and tumour-cell plasticity: lessons from melanoma. Nature Rev Cancer 3:411–421CrossRefGoogle Scholar
Reisdorff J, En-Nia A, Stefanidis I, Floege J, Lovett DH, Mertens PR (2002) Transcription factor Ets-1 regulates gelatinase a gene expression in mesangial cells. J Am Soc Nephrol 13:1568–1578PubMedCrossRefGoogle Scholar