In Vitro Radionuclide Therapy and In Vivo Scintigraphic Imaging of Alpha-Fetoprotein-Producing Hepatocellular Carcinoma by Targeted Sodium Iodide Symporter Gene Expression

Abstract

Purpose This study aimed to develop a gene expression targeting method for specific imaging and therapy of alpha-fetoprotein (AFP)-producing hepatocellular carcinoma (HCC) cells, using an adenovirus vector containing the human sodium/iodide symporter (hNIS) gene driven by an AFP enhancer/promoter. Methods The recombinant adenovirus vector, AdAFPhNIS (containing the hNIS gene driven by human AFP enhancer/ promoter) was prepared. After in vitro infection by the adenovirus, hNIS gene expression in AFP-producing cells and in AFP-nonproducing cells was investigated using 125 I uptake assay and semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). The killing effect of 131 I on AdAFPhNIS-infected HCC cells was studied using an in vitro clonogenic assay. In addition, tumor-bearing mice were intravenously injected with the adenovirus, and scintigraphic images were obtained. Results The expression of hNIS was efficiently demonstrated by 125 I uptake assay in AFP-producing cells, but not in AFP-nonproducing cells. AFP-producing HCC-targeted gene expression was confirmed at the mRNA level. Furthermore, in vitro clonogenic assay showed that hNIS gene expression induced by AdAFPhNIS infection in AFPproducing cells caused more sensitivity to 131 I than that in AFP-nonproducing cells. Injected intravenously in HuH-7 tumor xenografts mice by adenovirus, the functional hNIS gene expression was confirmed in tumor by in vivo scintigraphic imaging. Conclusions An AFP-producing HCC was targeted with an adenovirus vector containing the hNIS gene using the AFP enhancer/promoter in vitro and in vivo. These findings demonstrate that AFP-producing HCC-specific molecular imaging and radionuclide gene therapy are feasible using this recombinant adenovirus vector system.

Adenovirus-mediated expression of human sodium-iodide symporter gene permits in vivo tracking of adipose tissue-derived stem cells in a canine myocardial infarction model

“…After a 24-h incubation, the cells were assessed for NIS activity by radioiodine uptake at 37°C [17,18]. Briefly, the cells were incubated at 37°C for 120 min in 1 mL of Hank's Balanced Salt Solution (HBSS) containing 0.5% BSA, 3.7 kBq (0.1 μCi) of carrierfree 125 I-NaI, and 10 μmol/L of NaI to yield a specific activity of 740 MBq/mmol.…”

“…In the stem cell field, molecular imaging of cell tracking has been studied in a variety of ways, including direct cell labeling methods by using 18 F-fluorodeoxyglucose, 64 Cu-diacetyl-bis(N4-methylsemicarbazone), or iron oxide, and indirect methods using genetically transduced cells expressing reporter protein such as herpes simplex virus type 1 thymidine kinase (HSV1-tk), the sodium iodide symporter (NIS), or the firefly luciferase [3][4][5][6][7]. However, pre-clinical studies using optical imaging systems have limitations when applied to clinical practice [8].…”

Enhanced transferrin modified targeting delivery system via AFP promoter MR imaging reporter gene transfection

“…Alpha-fetoprotein (AFP) is a primitive embryonic protein, which is expressed positively in 80% of hepatocellular carcinoma and some embryonal tumors [25]. Therefore, as a serum tumor marker of HCC, its elevation often indicates the presence of HCC [25,26]. Utilize the specific expression of AFP in hepatocellular carcinoma to introduce the plasmid which concluded an AFP promoter and Fth as subsequent genes.…”

“…Utilize the specific expression of AFP in hepatocellular carcinoma to introduce the plasmid which concluded an AFP promoter and Fth as subsequent genes. Promote the expression of follow-up ferritin gene for early diagnosis [25,27].…”

Establishment of Animal Models with Orthotopic Hepatocellular Carcinoma

“…Investigation of HCC has been done conducted at the level of genes, ribonucleic acid (RNA), proteins, cells, and animals [10][11][12][13][14]. The resultant development of animal models and monitoring methods has improved the effectiveness of guidelines provided to researchers working with preclinical HCC models.…”