Chen Zhi-Yi, Liang Kun, Lin Yan, Yang Feng
Department of Medical Ultrasound, Key Laboratory for Major Obstetric Diseases of Guangdong Province, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China.
Int J Mol Sci. 2013 Jan 16;14(1):1763-77. doi: 10.3390/ijms14011763.
Apoptosis induction by short hairpin RNA (shRNA) expression vectors could be an efficient and promising strategy for cancer gene therapy. Ultrasound-targeted microbubble destruction (UTMD) is an appealing technique. In this study, we investigated the apoptosis induction and suppression of cell proliferation in vivo transfected by the UTMD-based shRNA delivery system. Nude mice with transplanted tumors of cervical cancer were randomly arranged into three groups: control group, plasmid injection and ultrasound (P + US), P + UTMD group. Expressions of Survivin and proliferating cell nuclear antigen (PCNA), Bcl-2, Bax, Caspase-3, Ki-67, nucleostemin (NS) were investigated by immunohistochemistry. Furthermore, microvessel density (MVD) was detected by CD34 protein expressions and apoptotic index (AI) was measured by TUNEL. As compared with those in the control and P + US groups, protein expressions of PCNA, Ki-67, Bcl-2, Survivin and NS in P + UTMD groups were down-regulated markedly, while those of Bax, Caspase-3 were up-regulated significantly (p < 0.05). MVD decreased significantly, whereas AI increased remarkably (p < 0.05). We suggested that UTMD-based shRNA delivery system could induce apoptosis and inhibit proliferation significantly, without causing any apparently adverse effect, representing a new, promising technology that would be used in the future gene therapy and research.
短发夹RNA（shRNA）表达载体诱导细胞凋亡可能是一种高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。在本研究中，我们调查了基于UTMD的shRNA递送系统体内转染后诱导细胞凋亡及抑制细胞增殖的情况。将移植有宫颈癌肿瘤的裸鼠随机分为三组：对照组、质粒注射加超声组（P + US）、P + UTMD组。通过免疫组织化学研究存活素、增殖细胞核抗原（PCNA）、Bcl-2、Bax、半胱天冬酶-3、Ki-67、核干细胞因子（NS）的表达。此外，通过CD34蛋白表达检测微血管密度（MVD），通过TUNEL检测凋亡指数（AI）。与对照组和P + US组相比，P + UTMD组中PCNA、Ki-67、Bcl-2、存活素和NS的蛋白表达明显下调，而Bax、半胱天冬酶-3的蛋白表达显著上调（p < 0.05）。MVD显著降低，而AI显著增加（p < 0.05）。我们认为基于UTMD的shRNA递送系统可显著诱导细胞凋亡并抑制增殖，且不会引起任何明显的不良反应，是一种可用于未来基因治疗和研究的新型、有前景的技术。
