Ultrasound-mediated destruction of gene-loaded microbubbles enhanced the transfection of phospholamban antisense RNA into cardiac myocytes
10.3760/cma.j.issn.1004-4477.2010.10.026
- VernacularTitle:超声破裂载基因微泡增强心肌细胞受磷蛋白反义RNA转染
- Author:
Guozhong WANG
;
Jinghua LIU
;
Shuzheng Lü
;
Chengjun GUO
;
Yingchuan ZHANG
;
Fei YUAN
;
Xiantao SONG
;
Yuan ZHOU
;
Changjiang GE
- Publication Type:Journal Article
- Keywords:
Ultrasonography;
Microbubbles;
RNA,antisense;
Transfection;
Phospholamban
- From:
Chinese Journal of Ultrasonography
2010;19(10):897-900
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore whether ultrasound-mediated microbubbles destruction could enhance anti-sense RNA transfection and expression. Methods Phospholamban antisense RNA eukaryon vector PcDNA 4. 1-asPLB was successfully constructed and it was transfected into cardiac myocytes by various methods including calcium phosphate precipitation, ultrasound exposure and ultrasound-mediated microbubbles destruction. The expression of PLB and sarcoplasmic retculum Ca2+ ATPase (SERCA2a) in cardiac myocytes was tested by RT-PCR and western blot. Results The transfection and expression of PcDNA 4. 1-asPLB increased significantly in cells treated with ultrasound-mediated microbubbles destruction compared to other transfer groups( P ＜0.05). The expression of PLB was inhibited specifically after cardiac myocytes were transfected with PcDNA 4. 1-asPLB. There was no change of PLB expression after cardiac myocytes transfected with PcDNA 4. 1 ( P ＜0.05). Though the expression of SERCA2a never exhibited any changes after PcDNA 4. 1-asPLB transfection, the PLB/SERCA2a ratio decreased markedly. Conclusions As a highly effective antisense RNA transfer method, ultrasound-mediated microbubbles destruction can enhance the transfection and expression of the PcDNA 4. 1-asPLB significantly. The PcDNA4. 1-asPLB transfection inhibits the expression of PLB and result in decrease of PLB/SERCA2a ratio in cardiac myocytes.
