Chen Zhi-Yi, Liang Kun, Sheng Xiu-Jie, Si-Tu Bing, Sun Xiao-Fang, Liu Jian-Qiao, Qiu Ri-Xiang, Zhang Hua, Li Yue-Wei, Zhou Xin-Xin, Yu Jiang-Xiu
Department of Medical Ultrasound, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, P.R. China.
Oncol Lett. 2012 May;3(5):1030-1036. doi: 10.3892/ol.2012.610. Epub 2012 Feb 15.
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.
短发夹RNA(shRNA)表达载体诱导细胞凋亡可能是一种有效且有前景的癌症基因治疗策略。超声靶向微泡破坏(UTMD)是一种有吸引力的技术;然而,几乎没有数据可证明该技术的可行性并对其方法进行优化。本研究的目的是优化该技术,并阐明其在体外对基因抑制和细胞凋亡诱导的影响。获取人宫颈癌细胞系并进行培养。构建shRNA载体,并检测UTMD技术是否适合将shRNA转染到细胞中。然后使用流式细胞术对细胞进行检测。结果显示,最佳照射参数可获得更高的转染效率,且不影响质粒DNA的完整性。我们得出结论,在最佳UTMD参数介导下,shRNA表达载体下调生存素可显著诱导细胞凋亡并导致细胞周期停滞,为进一步研究这种癌症治疗方法奠定了基础。