title: Evaluation of antitumor effects following tumor necrosis factor-α gene delivery using nanobubbles and ultrasound.
abstract: The antitumor effects of tumor necrosis factor (TNF-α) were evaluated following transfection of TNF-α plasmid DNA into solid mouse tumors using the nanobubbles (NBs) and ultrasound (US) gene delivery system. Murine breast carcinoma (EMT6) cells expressing luciferase (1 × 10(6) cells) were injected intradermally into the flanks of 6-7-week-old male SCID mice on day 0. Ten microliters of TNF-α (5 μg/μL) or TNF-α mock plasmid DNA (5 μg/μL) with/without NBs (15 μL) and saline was injected intratumorally in a total volume of 30 μL, and tumors were exposed to US (frequency, 1 MHz; intensity, 3.0 W/cm(2); duty cycle, 20%; number of pulses, 200; and exposure time, 60 s) on days 2, 4, 7, and 9. Changes in tumor size were measured with an in vivo bioluminescent imaging system and a mechanical caliper. Changes in tumor vessel area were quantified using contrast-enhanced US imaging with Sonazoid and a high frequency US imaging system (40 MHz) and immunohistochemistry (CD31). At the mRNA level, expression of TNF-α, caspase-3, and p53 were quantified using real-time quantitative RT-PCR. At the protein level, expression of caspase-3 and p53 were confirmed by immunohistochemistry. We show that repeated TNF-α gene delivery using NBs and US can lead to the local production of TNF-α. This results in antitumor effects, including activation of p53-dependent apoptosis, decrease in tumor vessel density, and suppression of tumor size. In this study, we showed the effectiveness of using NBs and US for TNF-α gene delivery into tumor cells.
authors.affiliation: Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan.
authors.forename: Sachiko
authors.initials: S
authors.lastname: Horie
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Watanabe
authors.forename: Masao
authors.initials: M
authors.lastname: Ono
authors.forename: Shiro
authors.initials: S
authors.lastname: Mori
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Kodama
mesh_terms: D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000096:biosynthesis / Q000235:genetics; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000627:therapeutic use*; D004354:Drug Screening Assays, Antitumor; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D008297:Male; D008325:Mammary Neoplasms, Experimental / Q000098:blood supply / Q000000981:diagnostic imaging / Q000473:pathology / Q000628:therapy*; D051379:Mice; D016513:Mice, SCID; D053769:Nanocapsules / Q000008:administration & dosage*; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use; D014409:Tumor Necrosis Factor-alpha / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics / Q000627:therapeutic use*; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis / Q000235:genetics; D014465:Ultrasonics; D014463:Ultrasonography
chemical_list: D004274:DNA, Recombinant; D053769:Nanocapsules; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha; D016159:Tumor Suppressor Protein p53; C505409:Casp3 protein, mouse; D053148:Caspase 3