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- Title
Fluorescence Microscopy Experimental Study of Cell Transfection Induced by Ultrasonic Cavitation.
- Authors
Zihong Liu; Hui You; Ping Zhang
- Abstract
The development of non-viral gene transfer methods will provide a valuable supplement for gene therapy. Ultrasonic cavitation is an effective method for cell transfection. Ultrasonic energy can produce a variety of biological effects through acoustic cavitation, cavitation bubble can make cell dead or cell membrane permeability changes, which is beneficial to gene transfer. Ultrasound cavitation for cell transfection has been confirmed in in vitro culture experiments and mammalian experiments. Which provides a promising clinical medical approach for the development of new gene therapies? The aim of this study was to investigate the efficiency and biosafety of transfection of HA2741 gene into human breast cancer cells by ultrasound cavitation. The experimental method is to culture human breast cancer cells in vitro, respectively conduct the treatment of simple ultrasound, simple contrast agent, simple HA2741, ultrasonic irradiation + HA2741, contrast agent + HA2741, contrast agent +HA2741 + ultrasound irradiation, liposome + HA2741, liposome + HA2741+ultrasonic irradiation, the acoustic emission intensity was -18.0~0 dB, and the sonication time was 5~240 seconds. After treatment, use the fluorescence microscope to observe the transfection rate and cell activity of HA2741, and use the immunohistochemistry to detect the expression of protein HER-2. The experimental results showed that the transfection rate of the contrast agent +HA2741+ultrasound group was as high as about 94.6%, which was much higher than other groups; the transfection efficiency was the highest when the sonic intensity was -3.0~-6.0dB and the irradiation was 30~60 seconds; and the transfection efficiency is higher, the expression of HER-2 in breast cancer cells is lower. The results show that the ultrasonic cavitation micro bubble contrast agent can improve the transfection efficiency, and has the advantages of safety and simplicity.
- Subjects
FLUORESCENCE microscopy; GENE transfection; BREAST cancer gene therapy; CAVITATION; GENETIC transformation; ULTRASONIC imaging; ACOUSTIC emission
- Publication
Acta Microscopica, 2019, Vol 28, Issue 6, p1851
- ISSN
0798-4545
- Publication type
Article