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Enhanced Expression of Adenovirus-Mediated Sodium Iodide Symporter Gene in MCF-7 Breast Cancer Cells with Retinoic Acid Treatment

¹ Department of Nuclear Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea; ² Department of Nuclear Medicine, Armed Forces Capital Hospital, Seongnam, Korea; ³ Department of Microbiology, University of Ulsan College of Medicine, Seoul, Korea; ⁴ Department of Otolaryngology–Head and Neck Surgery, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea; and ⁵ Institute for Biomacromolecules, University of Ulsan College of Medicine, Seoul, Korea

View larger version (49K): [in this window] [in a new window]   FIGURE 1.  Efficiency of adenovirus-mediated gene transfer in MCF-7 cells shown by X-Gal staining. Blue-colored cells represent expression of ß-galactosidase. Efficiency of gene transfer is >60% at 50 MOI. MI = mock infection.

View larger version (7K): [in this window] [in a new window]   FIGURE 2.  Effect of ATRA on growth inhibition of MCF-7 cells. Cellular proliferation was inhibited by ATRA in a dose-dependent manner over 10–8–10–6 mol/L concentration range (P < 0.05).

View larger version (9K): [in this window] [in a new window]   FIGURE 3.  Results of quantitative real-time RT-PCR. NIS mRNA expression was increased 2.1-fold with ATRA treatment in wild-type MCF-7 cells, whereas NIS expression increased by 118.5-fold in adenovirus-infected MCF-7 cells. Enhancement effect of ATRA is more marked in adenovirus-infected cells.

View larger version (29K): [in this window] [in a new window]   FIGURE 4.  Effect of ATRA treatment on NIS protein expression in MCF-7 cells assessed by Western blot analysis. Main immunoreactive protein in MCF-7 cells, 90 kDa, increased with ATRA treatment combined with adenoviral gene transfer. ATRA significantly increased expression in dose-dependent manner in cells infected with adenoviruses carrying NIS. However, wild-type MCF-7 cells showed only faint NIS expression despite ATRA treatment. hNIS = human NIS.

View larger version (25K): [in this window] [in a new window]   FIGURE 5.  MCF-7 cells with adenovirus-mediated NIS gene transfer showed prominent plasma membrane staining with intracellular immunoreactivity after ATRA treatment (A–D), whereas wild-type MCF-7 cells revealed only faint staining (not shown). (A) No ATRA. (B) 10–8 mol/L ATRA. (C) 10–7 mol/L ATRA. (D) 10–6 mol/L ATRA.

View larger version (24K): [in this window] [in a new window]   FIGURE 6.  MCF-7 cells with adenovirus-mediated NIS transfer showed increasing numbers of fluorescent positive cells after ATRA treatment (A–D), whereas wild-type MCF-7 cells showed only minimal increases in fluorescently labeled cells with ATRA treatment (not shown). (A) No ATRA. (B) 10–8 mol/L ATRA. (C) 10–7 mol/L ATRA. (D) 10–6 mol/L ATRA. FL2-H = 585/42 filter-height of fluorescence intensity.

View larger version (13K): [in this window] [in a new window]   FIGURE 7.  Effect of ATRA on radioiodine uptake in MCF-7 cells. 125I uptake in wild-type MCF-7 cells increased by 3.1-, 5.5-, and 7.6-fold with treatment of 10–8, 10–7, and 10–6 mol/L ATRA, respectively. Treatment with 0, 10–8, 10–7, and 10–6 mol/L ATRA in Rad-NIS–infected cells increased 125I uptake by 4.0-, 4.9-, 8.2-, and 27.6-fold greater than that in wild-type MCF-7 cells. Rad-NIS–infected MCF-7 cells with 10–6 mol/L ATRA treatment (245.0 ± 13.7 pmol/106 cells) is much greater than the sum of levels seen with ATRA treatment and Rad-NIS infection alone. RA = retinoic acid.