Effect of NET-1 siRNA conjugated sub-micron bubble complex combined with low-frequency ultrasound exposure in gene transfection
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Bolin Wu1, Xitian Liang1, Hui Jing1, Xue Han1, Yixin Sun1, Cunli Guo1, Ying Liu1 and Wen Cheng1
1Department of Ultrasound, Harbin Medical University Cancer Hospital, Nangang, Harbin, 150081, Heilongjiang Province, China
Wen Cheng, email: [email protected]
Keywords: sub-micron bubbles; low-frequency ultrasound; NET-1 siRNA; hepatocellular carcinoma; gene silencing
Received: April 21, 2017 Accepted: December 14, 2017 Published: December 23, 2017
The present study evaluated the effect of NET-1 siRNA-conjugated sub-micron bubble (SMB) complexes combined with low-frequency ultrasound exposure in gene transfection. The NET-1 gene was highly expressed level in SMMC-7721 human hepatocellular carcinoma cell line. The cells were divided into seven groups and treated with different conditions. The groups with or without low-frequency ultrasound exposure, groups of adherent cells, and suspension cells were separated. The NET-1 siRNA-conjugated SMB complexes were made in the laboratory and tested by Zetasizer Nano ZS90 analyzer. Flow cytometry was used to estimate the transfection efficiency and cellular apoptosis. Western blot and quantitative real-time polymerase chain reaction (qPCR) were used for the estimation of the protein and mRNA expressions, respectively. Transwell analysis determined the migration and invasion capacities of the tumor cells. The results did not show any difference in the transfection efficiency between adherent and suspension cells. However, the NET-1 siRNA-SMB complexes combined with low-frequency ultrasound exposure could enhance the gene transfection effectively. In summary, the NET-1 siRNA-SMB complexes appeared to be promising gene vehicle.
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