title: Enhanced antiproliferative effects of combination hexokinase II shRNA and NIS gene therapy on vascular smooth muscle cells.
abstract: BACKGROUND
  This study was designed to determine the antiproliferative effects of combination gene therapy using sodium iodide symporter (NIS)-based radioiodine and lentivirus-mediated short hairpin RNA (shRNA) against hexokinase II (HKII) on vascular smooth muscle cells (VSMCs).
  
  
  METHODS
  A7r5 rat VSMCs were stably transfected with a dual-expression vector of NIS and Fluc (A7r5-NL cells). Functional assessment was performed by radioiodine uptake assay, luciferase assay and confocal microscopy. After exposure to lentivirus-HKII-shRNA, the (18)F-FDG uptake test and HK activity assay were performed. The effects of combination therapy with (131)I and lentivirus-HKII-shRNA on VSMCs were assessed with an in vitro clonogenic assay. In vivo bioluminescence and nuclear imaging were undertaken using a xenografted mouse model.
  
  
  RESULTS
  In vitro functional assessment confirmed expression of NIS and Fluc genes in A7r5-NL, but not in parent A7r5 cells. Transfection of lentivirus-HKII-shRNA resulted in a significant decrease in messenger RNA expression of the HKII gene, (18)F-FDG uptake and HK activity. The cell survival rate of A7r5-NL decreased to 61.9% and 90.5% by single therapy with 7.4 MBq of (131)I or lentivirus-HKII-shRNA, respectively, and further decreased to 42.9% by combined therapy (P<.05). In vivo bioluminescent and gamma camera images clearly demonstrated optical signals and (99m)Tc pertechnetate uptake at the site of A7r5-NL cell inoculation in nude mice.
  
  
  CONCLUSIONS
  The enhanced antiproliferative effect on VSMCs was achieved by a combination of NIS-based radioiodine and lentivirus-mediated HKII shRNA gene therapy. Successful demonstration of in vivo dual reporter gene imaging assures the potential for further application in an animal model.
authors.affiliation: Department of Nuclear Medicine, School of Medicine, Kyungpook National University, Daegu 700-721, Republic of Korea.
authors.forename: Mi-Hye
authors.initials: MH
authors.lastname: Hwang
authors.forename: Sang-Woo
authors.initials: SW
authors.lastname: Lee
authors.forename: Jung Eun
authors.initials: JE
authors.lastname: Kim
authors.forename: Ho Won
authors.initials: HW
authors.lastname: Lee
authors.forename: Yong Hyun
authors.initials: YH
authors.lastname: Jeon
authors.forename: Jeoung-Hee
authors.initials: JH
authors.lastname: Ha
authors.forename: Byeong-Cheol
authors.initials: BC
authors.lastname: Ahn
authors.forename: Jaetae
authors.initials: J
authors.lastname: Lee
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D019788:Fluorodeoxyglucose F18 / Q000493:pharmacokinetics; D015316:Genetic Therapy / Q000379:methods*; D006593:Hexokinase / Q000235:genetics / Q000378:metabolism; D007457:Iodine Radioisotopes / Q000494:pharmacology*; D016086:Lentivirus / Q000235:genetics; D008156:Luciferases; D049408:Luminescent Agents; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D018613:Microscopy, Confocal; D009131:Muscle, Smooth, Vascular / Q000000981:diagnostic imaging / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000000981:diagnostic imaging / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011877:Radionuclide Imaging; D019275:Radiopharmaceuticals / Q000494:pharmacology; D051381:Rats; D013670:Sodium Pertechnetate Tc 99m / Q000493:pharmacokinetics; D027981:Symporters / Q000235:genetics / Q000378:metabolism; D000099209:Sodium-Iodide Symporters
chemical_list: D019788:Fluorodeoxyglucose F18; D006593:Hexokinase; D007457:Iodine Radioisotopes; D008156:Luciferases; D049408:Luminescent Agents; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D019275:Radiopharmaceuticals; D013670:Sodium Pertechnetate Tc 99m; D027981:Symporters; D000099209:Sodium-Iodide Symporters