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Introduction of plasmids into gastric cancer cells by endoscopic ultrasound

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Short hairpin RNA of frizzled-2 (shRNA-Fz2) suppresses the cell proliferation of gastric cancer cells. Endoscopic ultrasound (EUS) is considered a suitable method for the introduction of therapeutic plasmids into cells, since the device enables the access and realtime monitoring of gastric cancer tissues. In the present study, plasmids were introduced into cells by sonoporation, as evidenced by the production of H2O2. The production of H2O2 was measured by absorbance of a potassiumstarch solution irradiated with EUS. Luciferase activity was analyzed in the cells irradiated with EUS after the addition of a pMetLuc2-control in the media, and cell proliferation was analyzed using a 3(4,5dimethylthiazol2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2Htetrazolium inner salt assay after irradiation with EUS following the addition of shRNAFz2. Absorbance levels corresponding to free radical levels were found to be higher in the cells irradiated with EUS. Luciferase activities were found to be significantly higher in the transfected cells (plasmid with Lipofectamine LTX) than in untreated cells and were furthermore found to be higher in MKN45 cells irradiated for 0.5 min than in cells not subjected to irradiation. Luciferase activity was also found to be higher in MKN74 cells irradiated for 2 min than in cells that were not irradiated. Although the cell proliferation of the MKN45 cells tended to be suppressed by irradiation with EUS, this was nonsignificant suppression, while the cell proliferation of MKN74 cells was found to be suppressed by irradiation with 12 MHz for 2 min (P<0.05). In conclusion, plasmids were introduced into cultured gastric cancer cells by irradiation with EUS due to sonoporation, as evidenced by the production of H2O2; however, the efficiency of the plasmid introduction was low compared with a traditional transfection approach.
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Document Type: Research Article

Affiliations: 1: Department of Gastroenterology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 2840003, Japan 2: Department of Radiology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 2840003, Japan 3: Department of Neurology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 2840003, Japan 4: Department of Rheumatology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 2840003, Japan 5: Department of Pediatrics, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 2840003, Japan 6: Department of Neurosurgery, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 2840003, Japan

Publication date: May 1, 2017

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  • Oncology Letters is a monthly, peer-reviewed journal, available in print and online, that focuses on all aspects of clinical oncology, as well as in vitro and in vivo experimental model systems relevant to the mechanisms of disease.

    The principal aim of Oncology Letters is to provide the prompt publication of original studies of high quality that pertain to clinical oncology, chemotherapy, oncogenes, carcinogenesis, metastasis, epidemiology and viral oncology in the form of original research, reviews and case reports.
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