NSAIDs in neuroblastoma therapy

doi:10.1016/j.canlet.2005.01.058

Cancer Letters

Volume 228, Issues 1–2, 18 October 2005, Pages 195-201

https://doi.org/10.1016/j.canlet.2005.01.058 Get rights and content

Abstract

Cyclooxygenases (COX) catalyse the conversion of arachidonic acid to prostaglandins. COX-2 is upregulated in several adult epithelial cancers. In neuroblastoma it has been shown that the majority of primary tumours and cell lines express high levels of COX-2, whereas normal adrenal medullas from children do not express COX-2. Treatment of neuroblastoma cells with nonsteroidal anti-inflammatory drugs (NSAIDs), inhibitors of COX, induces caspase-dependent apoptosis via the intrinsic mitochondrial pathway. Established neuroblastoma xenografts in nude rats treated with the dual COX-1/COX-2 inhibitor, diclofenac, or the COX-2 specific inhibitor, celecoxib significantly inhibits neuroblastoma growth in vivo. In vitro, arachidonic acid and diclofenac synergistically induces neuroblastoma cell death. This effect is further pronounced when lipoxygenases is inhibited simultaneously. Proton MR-spectroscopy (¹H MRS) of neuroblastoma cells treated with COX-inhibitors demonstrates accumulation of polyunsaturated fatty acids and depletion of choline compounds. Thus, ¹H MRS, which can be performed with clinical MR-scanners, is likely to provide pharmacodynamic markers of neuroblastoma response to COX-inhibition.

Taken together, these data suggest the use of NSAIDs as a novel adjuvant therapy for children with neuroblastoma.

Introduction

Arachidonic acid is released from cellular phospholipids by phospholipaseA₂ and converted to prostaglandins by two cyclooxygenase enzymes, COX-1 and COX-2 [1]. COX-1 is constitutively expressed in most tissues, whereas inflammatory stimuli, hormones and mitogens induce COX-2 expression [1], [2]. Increased amount of COX-2 are found commonly in both premalignant and malignant tissues of epithelial origin in adults and has been implicated in resistance to apoptosis, promotion of cell proliferation, increased tumour invasiveness, induction of metastases and angiogenesis as well as decreased immune surveillance [2]. Non-steroidal anti-inflammatory drugs (NSAIDs) are potent inhibitors of cylooxygenases. Epidemiological studies show that use of NSAID is associated with a reduced risk of several adult malignancies [3]. Consistent with this, tumour formation and growth are reduced in COX-1 or COX-2 deficient animals [4], [5], [6]. Hence, numerous experimental, epidemiologic, and clinical studies suggest that NSAIDs, particularly highly selective COX-2 inhibitors, have promise as anticancer agents.

Section snippets

Expression of cyclooxygenase-2 in neuroblastoma

A majority of neuroblastoma tissues and cell lines as well as ganglioneuromas express high levels of COX-2. In our study, 27 of 28 neuroblastoma samples (96%) showed specific expression of COX-2 protein in the cytoplasm of the tumour cells. No COX-2 protein was detected in the surrounding non-malignant adrenal medulla tissues [7]. The neuroblastoma samples were from different biological subsets and at all clinical stages. Seven out of 28 tumours were MYCN-amplified whereas nine had

Treatment of neuroblastoma with NSAIDs in vitro

NSAIDs vary in their abilities to inhibit COX-1 or COX-2. Aspirin is a relatively selective inhibitor of COX-1 whereas most other conventional NSAIDs, such as ibuprofen, sulindac, indomethacin and diclofenac, inhibit COX-1 and COX-2 to the same extent. A third class of NSAIDs, designated coxibs by the World Health Organization that selectively inhibits COX-2 were developed to suppress prostanoid formation by COX-2 in inflammation while sparing the protective effects of COX-1 and its products

NSAIDs induce apoptosis of neuroblastoma cells in vitro

Studies investigating the potential mechanisms behind NSAID-induced cytotoxicity of neuroblastoma cells demonstrated that both the dual COX-1/COX-2 inhibitor diclofenac and the COX-2 inhibitor, celecoxib induces changes in the mitochondrial transmembrane potential. This results in the activation of procaspase-9 and procaspase-3 followed by induction of apoptosis. No activation of procaspase-8 or cleavage of BID is observed, suggesting that the intrinsic apoptotic pathway is involved in

Accumulation of arachidonic acid, a possible mechanism for NSAID-induced cell death of neuroblastoma cells

In comparison with healthy nervous tissue, neuroblastoma cells contain increased levels of arachidonic acid [17], which is the main substrate for eicosanoid biosynthesis catalysed by COX and lipoxygenases (LOX) [1], [18] (Fig. 2). Inhibition of both COX- and LOX- metabolic pathways induce a synergistic cytotoxicity of SH-SY5Y neuroblastoma cells in vitro, whereas exogenous addition of arachidonic acid alone stimulates neuroblastoma cells proliferation (P<0.001; Table 2). However, the addition

Proton magnetic resonance spectroscopy can be used to monitor the effects of NSAIDs in neuroblastoma

Proton magnetic resonance spectroscopy (¹H MRS), which allows clinical monitoring of tumour biochemistry, is particularly useful for analysis of intracellular lipids [25], including polyunsaturated fatty acids (PUFAs), of which arachidonic acid is the most abundant in vivo [26]. Moreover, ¹H MRS typically shows increased content of PUFAs and methylene groups of mobile lipids in cancer cells undergoing chemotherapy-induced apoptosis [25], [27], [28]. We have investigated the possibility of

NSAIDs effectively inhibits neuroblastoma growth in vivo

The effects of NSAIDs on neuroblastoma growth in vivo have been studied in athymic rats carrying SH-SY5Y xenografts. In two independent therapeutic experiments athymic rats carrying neuroblastoma xenografts were randomly assigned to continuously receive diclofenac in their drinking water (200 or 250 mg/litre) or received 10 days of treatment with celecoxib (10 mg once daily) administered through a gastric feeding tube. In both experiments, treatment was started at the appearance of palpable

Conclusions

Accumulating data show that COX-2 is expressed in neuroblastoma and NSAIDs induce apoptosis and inhibit growth of neuroblastoma both in vitro and in vivo [7]. Since NSAIDs are clinically available, show effects against epithelial cancers in adults [2], [3] and are well tolerated in children [37], [38], trials to evaluate their efficacy as an adjuvant therapy in children with neuroblastoma are warranted. Moreover, ¹H MRS provides biochemical markers for the response of neuroblastoma cells to

Acknowledgements

The Swedish Children's Cancer Foundation, Swedish Cancer Society, The Cancer Society of Stockholm.

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NSAIDs in neuroblastoma therapy

Abstract

Introduction

Section snippets

Expression of cyclooxygenase-2 in neuroblastoma

Treatment of neuroblastoma with NSAIDs in vitro

NSAIDs induce apoptosis of neuroblastoma cells in vitro

Accumulation of arachidonic acid, a possible mechanism for NSAID-induced cell death of neuroblastoma cells

Proton magnetic resonance spectroscopy can be used to monitor the effects of NSAIDs in neuroblastoma

NSAIDs effectively inhibits neuroblastoma growth in vivo

Conclusions

Acknowledgements

Cell

Lancet

Neurosci Lett.

Lancet Oncol.

J. Biol. Chem.

Thromb. Res.

Trends Pharmacol. Sci.

Neoplasia.

J. Biol. Chem.

J. Biol. Chem.

Cyclooxygenases: structural, cellular, and molecular biology

Annu. Rev. Biochem.

Cyclooxygenase 2: a molecular target for cancer prevention and treatment

Trends Pharmacol. Sci.

Nonsteroidal anti-inflammatory drugs as anticancer agents: mechanistic, pharmacologic, and clinical issues

J. Natl. Cancer Inst.

Genetic disruption of Ptgs-1, as well as Ptgs-2, reduces intestinal tumorigenesis in Min mice

Cancer Res.

Deficiency of either cyclooxygenase (COX)-1 or COX-2 alters epidermal differentiation and reduces mouse skin tumorigenesis

Cancer Res.

Cyclooxygenase-2 is expressed in neuroblastoma, and nonsteroidal anti-inflammatory drugs induce apoptosis and inhibit tumor growth in vivo

Cancer Res.

Increased expression of cyclooxygenase-2 in malignant pheochromocytomas

J. Clin. Endocrinol. Metab.

Cyclooxygenase-2 expression in pediatric sarcomas

Pediatr. Dev. Pathol.

Cyclooxygenase-2 expression does not correlate with outcome in osteosarcoma and rhabdomyosarcoma

J. Pediatr. Hematol. Oncol.