Combined RNA Interference of Hexokinase II and ¹³¹I-Sodium Iodide Symporter Gene Therapy for Anaplastic Thyroid Carcinoma

Abstract

The purpose of this study was to investigate the enhanced therapeutic effect of the combined use of shRNA (small hairpin RNA) therapy for the hexokinase II (HKII) gene and 131 I human sodium iodide symporter (hNIS) as a gene therapy for in vitro and in vivo treatment of anaplastic thyroid carcinoma cells (ARO) in an animal model. Methods: A recombinant lentivirus containing a plasmid with the hNIS gene driven by phosphoglycerate kinase promoter and green fluorescent protein (GFP) linked with an internal ribosome entry site sequence was produced. ARO cells were transfected with the virus and sorted by fluorescent activated cell sorting using GFP (ARO-NG). The messenger RNA expression of hNIS and GFP were evaluated with reverse-transcriptase polymerase chain reaction, and the function of hNIS was verified by 125 I uptake. The lentiviral vector expressing shRNA against HKII (Lenti-HKII shRNA) was constructed and used to infect ARO-NG cells. The effect of Lenti-HKII shRNA was evaluated by reverse-transcriptase polymerase chain reaction, 18 F-FDG uptake, and HK activity. An in vitro clonogenic assay was performed after Lenti-HKII shRNA therapy, 131 I therapy, and a combined therapy. The therapies were also applied in vivo to an animal model with an ARO-NG xenograft, and the effects were assessed with caliper measurements and 18 F-FDG PET. Results: ARO-NG cells showed an 125 I uptake 76-fold higher than the parent ARO cells. Compared with the uninfected ARO-NG cells, ARO-NG cells infected with Lenti-HKII shRNA had lower HKII messenger RNA expression, lower 18 F-FDG uptake, and HK activity. The proliferation of ARO-NG cells was inhibited by 131 I and Lenti-HKII shRNA therapies and further inhibited by the combined 131 I and Lenti-HKII shRNA therapy. Both the Lenti-HKII shRNA therapy and the 131 I therapy inhibited in vivo tumor growth in the tumor xenograft model. The combined Lenti-HKII shRNA and 131 I therapy resulted in a further decrease of tumor growth. Conclusion: Our results suggest that the combined HKII shRNA and 131 I therapy has a stronger antitumor effect than either the 131 I therapy or the HKII shRNA alone. Therefore, this combined therapy could be used as a powerful strategy for treating anaplastic thyroid carcinoma.

PI3K/Akt signaling mediated Hexokinase-2 expression inhibits cell apoptosis and promotes tumor growth in pediatric osteosarcoma

“…Consistent with these reports that inhibition of HK2 exhibits a tumor-suppressive role in thyroid cancer [29], hepatocellular carcinoma [30] and colon cancer [31], we found that silencing of HK2 expression results in increased cell apoptosis and reduced colony formation ability. However, the exact mechanism involved in HK2-mediated glucose metabolism and tumor growth remain largely unexplored.…”

In�vitro study on human umbilical cord mesenchymal stem cells transfected with lentivirus‑mediated hNIS‑EGFP dual reporter gene and co‑labeled with superparamagnetic iron oxide

“…Lentiviral vectors are also able to effectively transfect neuronal, hepatic, myocardial, tumor, endothelial and stem cells (37). For cells that are difficult to transfect, such as primary, stem and non-dividing cells, the lentiviral vector increases the transduction efficiency of target genes greatly, allowing for the incorporation of RNAi, cDNA and study reporter genes (38). The lentiviral packaging system used in the present study is a four-plasmid system, consisting of pRsc-REV, pMD1g-pRRE, pMD2G and interference plasmid.…”

Sodium Iodide Symporter for Nuclear Molecular Imaging and Gene Therapy: From Bedside to Bench and Back

“…Reducing the radionuclide dose for NIS gene therapy is able to reduce the adverse effects, but might lead to limited effectiveness 47. Combined treatment of radionuclide NIS gene therapy with other therapeutic approaches could be more efficient to improve therapeutic outcomes and can reduce adverse effects of radionuclide NIS gene therapy by reducing the radionuclide dose.…”