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Microbubble and Pluronic P85 Under Ultrasound Irradiation Mediated Gene Transfection to HepG2 Cell

Author: ZhangXiJun
Tutor: LiKaiYan
School: Huazhong University of Science and Technology
Course: Medical Imaging and Nuclear Medicine
Keywords: Pluronic Micro bubble Gene Transfection
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 4
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Abstract


Objective To investigate whether Pluronic P85 and plasmid EGFP which adhered on the microbubble (MB) could enhance the gene transfection and expression to HepG2 cell under ultrasound (US) irradiation.Methods Plasmid encoding enhanced green fluorescent protein (pEGFP) was used as a report gene. HepG2 cell and plasmids DNA with or without microbubble/P85 were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm2, 20 s, 20% duty cycle,PRF 100Hz) and after 24 h transfection rates and cell viability were assessed by fluorescence microscopy, fluorescence activated cell sorting (FACS) analysis, and trypan blue exclusion.Results The gene transfection to HepG2 cell under ultrasound irradiation was much higher compared to that without ultrasound irradiation. When they were exposed to ultrasound, microbubble alone or P85 alone expressed lower green fluorescent protein, microbubble plus P85 obtained the higher expression efficiency at 30% microbubble concentration without obvious decreasing of survival rate(55.73±3.32)%.Conclusions Microbubble with pluronic P85 could enhance the gene transfection under ultrasound irradiation, and it could deserve further attention and investigation. Objective To investigate the appropriate microbubble (MB) concentration with pluronic P85 could enhance the gene transfection and expression to HepG2 cell under ultrasound (US) irradiation.Methods Plasmid encoding enhanced green fluorescent protein (pEGFP) was used as a report gene. HepG2 cell and plasmids DNA with P85/different microbubble concentration were exposed to ultrasound (US parameters:1 MHz, 1.0 W/cm2, 20 s, 20% duty cycle and 100 Hz pulse recurrent frequency)and after 24 h transfection rates and cell viability were assessed by fluorescence microscopy, FACS and trypan blue exclusion.Results The transfection efficiency of pEGFP+US+P85 was (9.72±1.21)%,it could get higher transfection rate when we added different concentration of microbubble. With the concentration of microbubble increased, the transfection efficiency was added as well as the death rate. The approving transfection efficiency was got at 30% microbubble concentration, with ideal gene transfection efficiency(22.14±3.06)% and without obvious decreasing of survival rate (55.73±3.32)%.Conclusions Microbubble with pluronic P85 could enhance the gene transfection under ultrasound irradiation, and it got the approving transfection rate at 30% microbubble concentration.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Liver tumors
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