Ultrasonic Diagnosis Center, Northwest Women's and Children's Hospital, Xi'an 710061, China.
Department of Ultrasonic, Shanghai Rici Women's and Children's Hospital, Shanghai 200040, China.
Tissue Cell. 2022 Aug;77:101843. doi: 10.1016/j.tice.2022.101843. Epub 2022 May 27.
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.
超声靶向微泡破坏（UTMD）是一种新的基因治疗方法，它使用携带靶基因的超声和微泡来实现基因转染。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。使用数据库、qRT-PCR 和 Western blot 检测肿瘤组织、相邻组织和细胞中的 ANLN 水平。通过 MTT 测定确定 SF6 的最佳浓度。通过流式细胞术选择机械指数（MI）。在用脂质体或 UTMD 介导的脂质体转染后，通过细胞功能实验、qRT-PCR 和 Western blot 评估 CC 细胞的生物学行为和 EZH2 水平。利用 qRT-PCR 和 Western blot 检测上皮-间充质转化（EMT）相关标志物和细胞凋亡相关标志物的表达。选择 10%SF6 和 0.28 的 MI 进行后续测试。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 下调 CC 细胞中的 EZH2 表达。EZH2 过表达逆转了 UTMD-siANLN 对 EMT 和细胞凋亡相关标志物的调节作用。UTMD 可以提高 siANLN 转染到 CC 细胞中的效率，从而诱导 CC 细胞恶性表型的抑制，这可能成为 CC 基因治疗的新靶点。
