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超声微泡介导的 ANLN 沉默抑制 EZH2 表达缓解宫颈癌进展。

Ultrasound microbubble-mediated delivery of ANLN silencing-repressed EZH2 expression alleviates cervical cancer progression.

机构信息

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.

出版信息

Tissue Cell. 2022 Aug;77:101843. doi: 10.1016/j.tice.2022.101843. Epub 2022 May 27.

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。

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