The Study Of Transfection Efficiency Of P53 Gene Into Cervical Carcinoma Cell Line Hela With Ultrasound Microbubbles

PARTⅠOPTIMAL PARAMETER SELECTION FOR TRANSFERRING P53 GENE INTO CERVICAL CARCINOMA CELLS HELA BY ULTRASOUND MICROBUBBLESObjective:To study the optimal transfection parameter of P53 gene into cervical carcinoma cells Hela by ultrasound microbubbles. Methods: Hela cells growth characteristics was observed by morphology.The inhibitory effects of different ultrasound condition on Hela cells was determined by MTT assay.The resulting ultrasound promoted P53 to transfect Hela cells,and ultimately to determine the optimal ultrasound parameters.Results:1,The cell survival rate were respectively ( 81.19±2.00 ) %,(77.61±1.93)%,(61.97±1.86)% when exposing to ultrasound (0.5, 0.75,1W/cm2 for 30s ).The cell survival rate were respectively (84.75±0.75)%,(81.19±2.00)%,(69.56±0.32)% when exposing to ultrasound (10,30,60s for 0.5 W/cm2). Ultrasound inhibited cell growth as an intensity and exposure-duration dependent manner.2,Selected ultrasound condition promoted P53 to transfect Hela cells. With ultrasound time of 30s and intensity of 0.5W/cm2, Hela cells expressed more GFP than the other groups, with a rate of 7.1%, the difference has statistical significance.Conclusion: ultrasound microbubble could inhibit cervical cancer Hela cells in vitro. With ultrasound time of 30s and intensity of 0.5 W/cm2, ultrasound microbubble could maximize the promotion of P53 plasmid transfection into cervical cancer Hela cells.PARTⅡTHE STUDY OF TRANSFECTION EFFICIENCY OF P53 GENE INTO CERVICAL CARCINOMA CELL LINE HELA BY ULTRASOUND MICROBUBBLESObjective: To study the transfection efficiency of P53 into Hela cells with ultrasound microbubbles.Methods: Ultrasonic intensity of 0.5W/cm2 and consecutive irradiation time of 30s were used for transfection,Hela cells were divided into 5 groups: blank control , plasmid , plasmid +microbubble, plasmid + ultrasound, plasmid + ultrasound + microbubble.The transfection efficiency was detected by RT-PCR.Results: plasmid + ultrasound + microbubble group has obvious light strip, plasmid + ultrasound group has a weak band, that the former P53/β-actin optical density ratio was more than the latter, and the difference has statistical significance, while the remaining group could not be detected the target gene expression.Conclusion: At optimized conditions, ultrasound microbubble can promote wild-type P53 plasmid transfect into cervical cancer Hela cells; separate ultrasonic irradiation has a weaker role in promoting transfection.