Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of monoamine transporters in neuroblastoma cell lines: Correlations to meta-iodobenzylguanidine (MIBG) uptake and tyrosine hydroxylase gene expression

doi:10.1016/0959-8049(95)00039-L

European Journal of Cancer

Volume 31, Issue 4, 1995, Pages 586-590

https://doi.org/10.1016/0959-8049(95)00039-L Get rights and content

Abstract

Radiolabelled meta-iodobenzylguanidine (MIBG) has been widely used in scintigraphy and targeted radiotherapy in patients with neuroblastoma. Recently, it has been demonstrated that MIBG is incorporated into neuroblastoma cells by the noradrenaline transporter. In vitro experiments on SK-N-SH human neuroblastoma cells performed in the present study showed that uptake of MIBG is inhibited by noradrenaline, more so by dopamine and to a lesser extent, by serotonin, indicating that the respective transporters may also contribute to MIBG uptake. However, neither dopamine nor serotonin transporter gene expression was detected. Noradrenaline transporter gene expression was found in 4 of 6 investigated cell lines, which correlated with specific MIBG uptake. Furthermore, an inverse correlation of noradrenaline transporter and tyrosine hydroxylase gene expression, the key regulatory enzyme of catecholamine synthesis, was observed. These data show that MIBG is specifically incorporated only in neuroblastoma cells in which there is noradrenaline transporter gene expression. Furthermore, the catecholamine status in neuroblastoma cells is regulated by a coordinate expression of the key elements of catecholamine synthesis and reuptake systems.

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For imaging of neuroblastoma and phaeochromocytoma, [¹²³I]meta-iodobenzylguanidine ([¹²³I]mIBG) is routinely used, whereas [¹⁸F]6-fluorodopamine ([¹⁸F]6-FDA) is sporadically applied for positron emission tomography in pheochromocytoma. Both substances are taken up by catecholamine transporters (CATs). In competition, some other cell types are able to take up catecholamines and related compounds probably by organic cation (OCT) [extraneuronal monoamine (EMT)] transporters (OCT1, OCT2, OCT3=EMT). In this study, we investigated the uptake of radioiodine-labeled meta-iodobenzylguanidine (mIBG) as well as [³H]dopamine (mimicring 6-fluorodopamine) and [³H]noradrenaline. SK-N-SH (neuroblastoma) and PC-12 (phaeochromocytoma) cells were used and compared with HEK-293 cells transfected with OCT1, OCT2 and OCT3, respectively. In order to gain a more selective uptake in CAT expressing tumor cells, different specific inhibitors were measured. Uptake of mIBG into OCT-expressing cells was similar or even better as into both CAT-expressing cell lines, whereas dopamine and noradrenaline uptake was much lower in OCT-expressing cells. In presence of corticosterone (f.c. 10⁻⁴ M], catecholamine and mIBG uptake into SK-N-SH and PC-12 cells was only slightly reduced. In contrast, this process was significantly inhibited in OCT2 and OCT3 transfected HEK-293 as well as in Caki-1 cells, which naturally express OCT3. We conclude that the well-known corticosteroid corticosterone might be used in combination with [¹⁸F]6-FDA or [¹²³I]mIBG to improve specific imaging of neuroblastoma and pheochromocytoma and to reduce irradiation dose to nontarget organs in [¹³¹I]mIBG treatment.

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PaperReverse transcriptase-polymerase chain reaction (RT-PCR) analysis of monoamine transporters in neuroblastoma cell lines: Correlations to meta-iodobenzylguanidine (MIBG) uptake and tyrosine hydroxylase gene expression

Abstract

Lancet

Mol BrainRes

Brain Res

Radiolabelied adrenergic neuron-blocking agents: adrenomedullary imaging with [131I]iodobenzylguanidine

J Nucl Med

Uptake and storage of m-iodobenzylguanidine are frequent neuronal functions of human neuroblastoma cell lines

Cancer Res

Iodine-131 MIBG scintigraphy of neuroendocrine tumors other than pheochromocytoma and neuroblastoma

J Nucl Med

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Phase III study of iodine 131 metaiodobenzylguanidine in chemoresistant neuroblastoma: a United Kingdom Children's Cancer Study Group investigation

J Clin Oncol

Comparison of the sodium dependency of uptake of meta-iodobenzylguanidine and norepinephrine into cultured bovine adrenomedullary cells

Mol Pharmac

Specific uptake of m-[125I]iodobenzylguanidine in the human neuroblastoma cell line SK-N-SH

Cancer Res

Effect of uptake-one inhibitors on the uptake of norepinephrine and metaiodobenzylguanidine

J Nucl Med

Meta-iodobenzylguanidine (MIBG) uptake and storage in the human neuroblastoma cell line SK-N-BE(2C)

Int J Cancer

Expression cloning of a cocaine- and antidepressant sensitive human noradrenaline transporter

Nature

Paper
Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of monoamine transporters in neuroblastoma cell lines: Correlations to meta-iodobenzylguanidine (MIBG) uptake and tyrosine hydroxylase gene expression

Phase $I II$ study of iodine 131 metaiodobenzylguanidine in chemoresistant neuroblastoma: a United Kingdom Children's Cancer Study Group investigation