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Am J Nucl Med Mol Imaging 2012;2(4):483-498
Original Article
Efficient multicistronic co-expression of hNIS and hTPO in prostate cancer cells for
nonthyroidal tumor radioiodine therapy
Guoquan Li*, Lei Xiang*, Weidong Yang, Zhe Wang, Jing Wang, Kai Chen
Department of Nuclear Medicine, Xijing Hospital, The Fourth Military Medical University, Xi’an, Shaanxi, China, 710032; Molecular
Imaging Center, Department of Radiology, Keck School of Medicine, University of Southern California, CA 90033, USA; Department of
Pediatrics, Shaanxi Maternal and Child Care Service Center, Xi’an, Shaanxi, China, 710003. *These authors contributed equally to
this work.
Received August 6, 2012; Accepted August 24, 2012; Epub October 15, 2012; Published October 30, 2012
Abstract: Radioiodine therapy has proven to be a safe and effective approach in the treatment of differentiated thyroid cancer. Similar
treatment strategies have been exploited in nonthyroidal malignancies by transfecting hNIS gene into tumor cells or xenografts.
However, rapid radioiodine efflux is often observed after radioiodine uptake, limiting the overall antitumor effects. In this study, we
aimed at constructing multicistronic co-expression of hNIS and hTPO genes in tumor cells to enhance the radioiodine uptake and
prolong the radioiodine retention. Driven by the cytomegalovirus promoter, hNIS and hTPO were simultaneously inserted into the
expression cassette of adenoviral vector. An Ad5 viral vector (Ad-CMV-hTPO-T2A-hNIS) was assembled as a gene therapy vehicle by
Gateway technology and 2A method. The co-expression of hNIS and hTPO genes was confirmed by a double-label
immunofluorescence assay. The radioiodine (125I) uptake and efflux effects induced by co-expression of hNIS and hTPO genes
were determined in transfected and non-transfected PC-3 cells. Significantly higher uptake (6.58 ± 0.56 fold, at 1 h postincubation)
and prolonged retention (5.47 ± 0.36 fold, at 1 h of cell efflux) of radioiodine (125I) were observed in hNIS and hTPO co-expressed
PC-3 cells as compared to non-transfected PC-3 cells. We concluded that the new virus vector displayed favorable radioiodine
uptake and retention properties in hNIS-hTPO transfected PC-3 cells. Our study will provide valuable information on improving the
efficacy of hNIS-hTPO co-mediated radioiodine gene therapy. (ajnmmi1208002).
Keywords: Gene therapy, prostate cance, hNIS, hTPO, gateway cloning system
Address all correspondence to:
Dr. Jing Wang
Department of Nuclear Medicine, Xijing Hospital
The Fourth Military Medical University
Xi’an, Shaanxi, China, 710032.
Tel: +86 029 84775449; Fax: +86 029 81230242
E-mail: wangjing@fmmu.edu.cn;
Dr. Kai Chen
Molecular Imaging Center
Department of Radiology, Keck School of Medicine
University of Southern California, CA 90033, USA.
Tel: +1323 442 3858; Fax: +1 323 442 3253
E-mail: chenkai@usc.edu.
Original Article
Efficient multicistronic co-expression of hNIS and hTPO in prostate cancer cells for
nonthyroidal tumor radioiodine therapy
Guoquan Li*, Lei Xiang*, Weidong Yang, Zhe Wang, Jing Wang, Kai Chen
Department of Nuclear Medicine, Xijing Hospital, The Fourth Military Medical University, Xi’an, Shaanxi, China, 710032; Molecular
Imaging Center, Department of Radiology, Keck School of Medicine, University of Southern California, CA 90033, USA; Department of
Pediatrics, Shaanxi Maternal and Child Care Service Center, Xi’an, Shaanxi, China, 710003. *These authors contributed equally to
this work.
Received August 6, 2012; Accepted August 24, 2012; Epub October 15, 2012; Published October 30, 2012
Abstract: Radioiodine therapy has proven to be a safe and effective approach in the treatment of differentiated thyroid cancer. Similar
treatment strategies have been exploited in nonthyroidal malignancies by transfecting hNIS gene into tumor cells or xenografts.
However, rapid radioiodine efflux is often observed after radioiodine uptake, limiting the overall antitumor effects. In this study, we
aimed at constructing multicistronic co-expression of hNIS and hTPO genes in tumor cells to enhance the radioiodine uptake and
prolong the radioiodine retention. Driven by the cytomegalovirus promoter, hNIS and hTPO were simultaneously inserted into the
expression cassette of adenoviral vector. An Ad5 viral vector (Ad-CMV-hTPO-T2A-hNIS) was assembled as a gene therapy vehicle by
Gateway technology and 2A method. The co-expression of hNIS and hTPO genes was confirmed by a double-label
immunofluorescence assay. The radioiodine (125I) uptake and efflux effects induced by co-expression of hNIS and hTPO genes
were determined in transfected and non-transfected PC-3 cells. Significantly higher uptake (6.58 ± 0.56 fold, at 1 h postincubation)
and prolonged retention (5.47 ± 0.36 fold, at 1 h of cell efflux) of radioiodine (125I) were observed in hNIS and hTPO co-expressed
PC-3 cells as compared to non-transfected PC-3 cells. We concluded that the new virus vector displayed favorable radioiodine
uptake and retention properties in hNIS-hTPO transfected PC-3 cells. Our study will provide valuable information on improving the
efficacy of hNIS-hTPO co-mediated radioiodine gene therapy. (ajnmmi1208002).
Keywords: Gene therapy, prostate cance, hNIS, hTPO, gateway cloning system
Address all correspondence to:
Dr. Jing Wang
Department of Nuclear Medicine, Xijing Hospital
The Fourth Military Medical University
Xi’an, Shaanxi, China, 710032.
Tel: +86 029 84775449; Fax: +86 029 81230242
E-mail: wangjing@fmmu.edu.cn;
Dr. Kai Chen
Molecular Imaging Center
Department of Radiology, Keck School of Medicine
University of Southern California, CA 90033, USA.
Tel: +1323 442 3858; Fax: +1 323 442 3253
E-mail: chenkai@usc.edu.
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