Optimization and apoptosis induction by RNAi with UTMD technology in vitro.
Apoptosis induction by short hairpin RNA (shRNA) expression vectors may be an efficient and promising strategy for cancer gene therapy. Ultrasound-targeted microbubble destruction (UTMD) is an appealing technique; however, there few data are available to demonstrate the feasibility and to optimize the methodology for this technology. The aim of this study was to optimize this technique and to elucidate the effects on gene inhibition and apoptosis induction in vitro. Human cervical cancer cell lines were obtained and cultured.shRNA vectors were constructed, and the UTMD technique was examined to determine whether or not it was suitable for shRNA transfection into cells. Cells were then examined using flow cytometry. The results revealed that the optimal irradiation parameters obtained higher transfection efficiency and did not affect the integrity of plasmid DNA. We concluded that survivin downregulation with shRNA expression vectors, mediated by the optimal UTMD parameters, markedly induced cell apoptosis and caused cell cycle arrest, laying a foundation for further investigation of this cancer therapy.
Department of Medical Ultrasound, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, P.R. China.
Zhi-Yi
ZY
Chen
Kun
K
Liang
Xiu-Jie
XJ
Sheng
Bing
B
Si-Tu
Xiao-Fang
XF
Sun
Jian-Qiao
JQ
Liu
Ri-Xiang
RX
Qiu
Hua
H
Zhang
Yue-Wei
YW
Li
Xin-Xin
XX
Zhou
Jiang-Xiu
JX
Yu