Article Figures & Data
Figures
- FIGURE 1.
In vivo experimental schedule of combined Lenti-HKII shRNA and 131I therapy. BALB/c nude mice were challenged with 5 × 106 ARO-NG cells by subcutaneous injection into right hind limbs and then were treated with PBS, Lenti-scramble shRNA, or Lenti-HKII shRNA by intratumoral injection on days 1 and 4. 131I was intraperitoneally injected during initial administration of Lenti-HKII shRNA therapy. Tumor size was measured with calipers every week, and tumor volume was calculated using formula m12 × m2 × 0.5236 (where m1 represents length of short tumor axis and m2 represents that of long tumor axis) until day 21 after therapy.
- FIGURE 2.
Immunohistochemistry with anti-hNIS antibody in ARO-NG cells using confocal microscopy. Cell nuclei were visualized with 4′,6-diamidino-2-phenylindole staining (blue). GFP expression (green) was observed throughout cells, and hNIS expression was mainly detected on cell membrane (red). Merged image showed true location of hNIS in cells, which was expected to be on cell membrane. Parent ARO cells did not show GFP or hNIS staining. DPAI = 4′,6-diamidino-2-phenylindole.
- FIGURE 3.
In vitro 18F-FDG uptake measured 72 h after Lenti-HKII shRNA or Lenti-scramble shRNA transfection in ARO-NG cells. Cells transfected with Lenti-HKII shRNA showed lower 18F-FDG uptake than cells treated with PBS or Lenti-scramble shRNA. *Mean P < 0.05.
- FIGURE 4.
In vitro clonogenic assay for cell survival on day 3 after Lenti-HKII shRNA or on day 7 after 131I therapy or combined Lenti-HKII shRNA and 131I therapy. Clone-forming ability of ARO-NG cells was inhibited 58% ± 8.1% by 131I or 64.5% ± 8.4% by Lenti-HKII shRNA therapy alone and further inhibited 29% ± 8.8% by combined 131I and Lenti-HKII shRNA therapy. *Mean P < 0.05.
- FIGURE 5.
(A) In vivo effects of Lenti-HKII shRNA and 131I therapy on ARO-NG tumor xenografts in animal model. Tumor volumes of all therapy groups were significantly smaller than those of control and Lenti-scramble shRNA therapy groups. *Mean P < 0.05. †Mean P < 0.001. (B) Tumor volumes of 131I therapy group were significantly smaller than those of Lenti-HKII shRNA therapy group on day 21. Tumor volumes of combined therapy group were significantly decreased, compared with other individual therapy groups on days 14 and 21. *Mean P < 0.05.
- FIGURE 6.
(A) 18F-FDG small-animal PET was performed in vivo before and at 7, 14, and 21 d after Lenti-HKII shRNA, 131I, or combined therapies. (B) Quantitated %ID/g data of 18F-FDG uptake. 18F-FDG uptake of ARO-NG tumor xenografts was inhibited by Lenti-HKII shRNA, 131I therapy, and combined therapies. Lenti-HKII shRNA therapy group showed gradual increase of tumor 18F-FDG uptake on days 14 and 21, compared with that of day 7. 131I therapy group showed similar levels of tumor 18F-FDG uptake on days 14 and 21, compared with day 7. Combined therapy group showed lowest tumor 18F-FDG uptake on days 14 and 21, compared with other groups receiving individual therapies. *Mean P < 0.05. Arrows = location of ARO-NG tumors.
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