Cotransfecting norepinephrine transporter and vesicular monoamine transporter 2 genes for increased retention of metaiodobenzylguanidine labeled with iodine 131 in malignant hepatocarcinoma cells

Abstract

Norepinephrine transporter (NET) transfection leads to significant uptake of iodine-131-labeled metaiodobenzylguanidine (¹³¹I-MIBG) in non-neuroendocrine tumors. However, the use of ¹³¹I-MIBG is limited by its short retention time in target cells. To prolong the retention of ¹³¹I-MIBG in target cells, we infected hepatocarcinoma (HepG2) cells with Lentivirus-encoding human NET and vesicular monoamine transporter 2 (VMAT2) genes to obtain NET-expressing, NET-VMAT2-coexpressing, and negative-control cell lines. We evaluated the uptake and efflux of ¹³¹I-MIBG both in vitro and in vivo in mice bearing transfected tumors. NET-expressing and NET-VMAT2-coexpressing cells respectively showed 2.24 and 2.22 times higher ¹³¹I-MIBG uptake than controls. Two hours after removal of ¹³¹I-MIBG-containing medium, 25.4% efflux was observed in NET-VMAT2-coexpressing cells and 38.6% in NET-expressing cells. In vivo experiments were performed in nude mice bearing transfected tumors; results revealed that NET-VMAT2-coexpressing tumors had longer ¹³¹I-MIBG retention time than NET-expressing tumors. Meanwhile, NET-VMAT2-coexpressing and NET-expressing tumors displayed 0.54% and 0.19%, respectively, of the injected dose per gram of tissue 24 h after ¹³¹I-MIBG administration. Cotransfection of HepG2 cells with NET and VMAT2 resulted in increased ¹³¹I-MIBG uptake and retention. However, the degree of increase was insufficient to be therapeutically effective in target cells.

Keywords

norepinephrine transporter / vesicular monoamine transporter 2 / ¹³¹I-MIBG / gene therapy / lentivirus vector

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Acknowledgements

We thank Yuanyou Yang, PhD, for helping in the preparation of ¹³¹I-MIBG. This study was funded by the National Natural Science Foundation of China (No. 81271602).

Compliance with ethics guidelines

Yanlin Zhao, Xiao Zhong, Xiaohong Ou, Huawei Cai, Xiaoai Wu, and Rui Huang declare no conflict of interest that would prejudice their impartiality. This article does not contain any studies with human participants. All institutional and national guidelines for the care and use of laboratory animals were followed.

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2017 Higher Education Press and Springer-Verlag Berlin Heidelberg