Abstract
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.
Highlights
Cancer cannot be cured using the traditional treatments such as surgery, radiotherapy, and chemotherapy
The present study evaluated the effect of neuroepithelial transforming protein 1 (NET-1) siRNA-conjugated sub-micron bubble (SMB) complexes combined with low-frequency ultrasound exposure in gene transfection
These results proved that NET-1 siRNA-SMB complexes and low-frequency ultrasound (LFUS) exposure could deliver NET-1 siRNA to SMMC-7721 cells, exhibiting high-performance and inhibit the NET-1 mRNA and protein expressions
Summary
Cancer cannot be cured using the traditional treatments such as surgery, radiotherapy, and chemotherapy. Hepatocellular carcinoma (HCC) is the sixth most common cancer and the third most common cause of cancer-related deaths worldwide [2, 3], standard chemotherapy shows a poor treatment efficacy [4]. Gene silencing therapy has been developed as a novel therapeutic strategy in recent years. Gene silencing and delivery have been demonstrated as promising methods for the treatment of cancer [5, 6]. The low transfection efficiency to specific tissues or organs impeded its wide clinical applications. Low-frequency ultrasound (LFUS) could enhance the transient permeability of plasma membranes to facilitate the uptake [7]
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