Abstract：
Ultrasound-targeted microbubble destruction (UTMD) is a new gene therapy method that uses ultrasound and microbubbles carrying target genes to achieve gene transfection. However, whether UTMD-mediated ANLN silencing transfection helps to restrain the growth of cervical cancer (CC) is obscure. ANLN level in tumor tissues, adjacent tissues, and cells was tested using the database, qRT-PCR, and western blot. The optimal concentration of SF6 was determined by MTT assay. Mechanical index (MI) was selected by flow cytometry. After transfection with liposome or UTMD-mediated liposome, cell function experiments, qRT-PCR, and western blot were employed to assess CC cell biological behaviors and EZH2 level. Epithelial-mesenchymal transition (EMT)-related marker and apoptosis-related marker expressions were examined utilizing qRT-PCR and western blot. 10% SF6 and MI of 0.28 were selected for subsequent tests. ANLN was highly expressed in CC and cells. The transfection efficiency of the UTMD-siANLN group was higher than that of the L-siANLN group. Moreover, the repression of UTMD-siANLN on CC cell malignant phenotypes was stronger than L-siANLN. UTMD-siANLN attenuated EZH2 expression in CC cells. The modulatory role of UTMD-siANLN on EMT- and apoptosis-related markers was reversed by EZH2 overexpression. UTMD can improve the efficiency of siANLN transfection into CC cells to induce suppression of CC cell malignant phenotypes, which may become a new target of gene therapy for CC.
摘要：
超声靶向微泡破坏(UTMD)是利用超声和携带靶基因的微泡实现基因转染的一种新的基因治疗方法。然而,UTMD介导的ANLN沉默转染是否有助于抑制宫颈癌（CC）的生长尚不清楚。肿瘤组织中的ANLN水平，邻近组织，使用数据库测试细胞，qRT-PCR,和westernblot.MTT法测定SF6的最佳浓度。通过流式细胞术选择机械指数(MI)。用脂质体或UTMD介导的脂质体转染后，细胞功能实验,qRT-PCR,采用免疫印迹法评估CC细胞生物学行为和EZH2水平。利用qRT-PCR和westernblot检测上皮-间质转化(EMT)相关标志物和凋亡相关标志物的表达。选择10%SF6和MI为0.28用于后续测试。ANLN在CC和细胞中高表达。UTMD-siANLN组的转染效率高于L-siANLN组。此外，UTMD-siANLN对CC细胞恶性表型的抑制作用强于L-siANLN。UTMD-siANLN减弱了CC细胞中的EZH2表达。UTMD-siANLN对EMT和凋亡相关标志物的调节作用被EZH2过表达逆转。UTMD可以提高siANLN转染CC细胞的效率，诱导CC细胞恶性表型的抑制，可能成为CC基因治疗的新靶点。
