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Gene Expression Profiles, Histologic Analysis, and Imaging of Squamous Cell Carcinoma Model Treated with Focused Ultrasound Beams

¹ Department of Radiology, Lucas MRS Research Center, Stanford University School of Medicine, Stanford, CA, 94305.
² Present address: Department of Clinical Radiology, Clinic of Grosshadern, Ludwig Maximilians University, Marchioninistrasse 15, Munich 81377, Germany.

View larger version (135K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 1 —Modified Sonablate 200 (Focus Surgery) with ultrasound probe. Photograph shows experimental setup of single-component ultrasound transducer, high-frequency amplifier, diagnostic and therapeutic ultrasound probe, and experimental water bath.

View larger version (133K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2A —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Control images; no application of high-intensity focused ultrasound (HIFU). Unenhanced T1-weighted images. In unenhanced T1-weighted image (A), no contrast uptake was seen as sign of devascularization.

View larger version (92K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2B —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Control images; no application of high-intensity focused ultrasound (HIFU). Unenhanced T1-weighted images. In unenhanced T1-weighted image (A), no contrast uptake was seen as sign of devascularization.

View larger version (96K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2C —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Control images; no application of high-intensity focused ultrasound (HIFU). Unenhanced T1-weighted images. In unenhanced T1-weighted image (A), no contrast uptake was seen as sign of devascularization.

View larger version (84K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2D —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Images after application of continuous-wave HIFU. Unenhanced T1-weighted image is D; no contrast material uptake (arrow) on contrast-enhanced T1-weighted image (E) was seen as sign of devascularization. On T2-weighted image (F), treated tissue shows higher signal intensity (arrow).

View larger version (116K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2E —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Images after application of continuous-wave HIFU. Unenhanced T1-weighted image is D; no contrast material uptake (arrow) on contrast-enhanced T1-weighted image (E) was seen as sign of devascularization. On T2-weighted image (F), treated tissue shows higher signal intensity (arrow).

View larger version (118K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2F —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Images after application of continuous-wave HIFU. Unenhanced T1-weighted image is D; no contrast material uptake (arrow) on contrast-enhanced T1-weighted image (E) was seen as sign of devascularization. On T2-weighted image (F), treated tissue shows higher signal intensity (arrow).

View larger version (86K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2G —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Images after pulsed-wave HIFU. No differences in signal intensities were seen between untreated tumors and tumors treated with pulsed-wave HIFU.

View larger version (94K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2H —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Images after pulsed-wave HIFU. No differences in signal intensities were seen between untreated tumors and tumors treated with pulsed-wave HIFU.

View larger version (86K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 2I —MRI of SCCVII (squamous cell carcinoma) tumors. From left to right, images are unenhanced T1-weighted, contrast-enhanced T1-weighted, and T2-weighted. Images after pulsed-wave HIFU. No differences in signal intensities were seen between untreated tumors and tumors treated with pulsed-wave HIFU.

View larger version (180K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3A —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. Untreated SCCVII tumors show few apoptotic cells in TUNEL staining (B) and no stain for heat shock protein 70 (HSP70) (C).

View larger version (161K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3B —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. Untreated SCCVII tumors show few apoptotic cells in TUNEL staining (B) and no stain for heat shock protein 70 (HSP70) (C).

View larger version (187K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3C —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. Untreated SCCVII tumors show few apoptotic cells in TUNEL staining (B) and no stain for heat shock protein 70 (HSP70) (C).

View larger version (139K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3D —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. After application of continuous-wave high-intensity focused ultrasound (HIFU), necrosis in tumor (arrows) was seen. In TUNEL staining (E), apoptotic cells and in immunohistochemistry, positive stain for HSP70 (F) was seen.

View larger version (101K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3E —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. After application of continuous-wave high-intensity focused ultrasound (HIFU), necrosis in tumor (arrows) was seen. In TUNEL staining (E), apoptotic cells and in immunohistochemistry, positive stain for HSP70 (F) was seen.

View larger version (125K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3F —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. After application of continuous-wave high-intensity focused ultrasound (HIFU), necrosis in tumor (arrows) was seen. In TUNEL staining (E), apoptotic cells and in immunohistochemistry, positive stain for HSP70 (F) was seen.

View larger version (170K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3G —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. In pulsed-wave mode, note many apoptotic cells (arrow, H) in TUNEL staining (H) but no stain for HSP70 (I) in immunohistochemistry.

View larger version (120K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3H —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. In pulsed-wave mode, note many apoptotic cells (arrow, H) in TUNEL staining (H) but no stain for HSP70 (I) in immunohistochemistry.

View larger version (142K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 3I —Histology of SCCVII (squamous cell carcinoma) tumors. From left to right, images are stained with H and E, terminal deoxynucleotidyl transferase-mediated dUTP (deoxyuridine triphosphate) nick-end labeling (TUNEL), and HSP70. In pulsed-wave mode, note many apoptotic cells (arrow, H) in TUNEL staining (H) but no stain for HSP70 (I) in immunohistochemistry.

View larger version (88K): [in this window] [in a new window] [as a PowerPoint slide]   Fig. 4 —Western blot analysis of heat shock protein 70 (HSP70) expression in SCCVII (squamous cell carcinoma) cells. Lane 1: marker proteins (M), Bio-Rad Protein Assay (Bio-Rad); lane 2: positive heat shock cell lysate (pos C), product # LYC-HL101 (StresssGene); lane 3: SCCVII cell lysate not exposed to high-intensity focused ultrasound (HIFU) treatment (notx); lane 4: continuous-wave HIFU-treated SCCVII tumor cells harvested directly after application (cw); lane 5: continuous-wave HIFU-treated SCCVII tumor cells harvested after 4 hours (cw-4h); lane 6: short-pulse wave HIFU-treated SCCVII tumor cells harvested after 4 hours (pw-4h). Each sample contained 100 µg of proteins from tissue lysates prepared as described.

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