Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Jinan University, 613 W. Huangpu Avenue, Guangzhou City, Guangdong Province, 510630, China; Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Bengbu Medical College, 287 Changhuai Road, Longzihu District, Bengbu City, Anhui Province, 233000, China.
Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Bengbu Medical College, 287 Changhuai Road, Longzihu District, Bengbu City, Anhui Province, 233000, China.
Tissue Cell. 2022 Feb;74:101719. doi: 10.1016/j.tice.2021.101719. Epub 2021 Dec 23.
Ultrasound-targeted microbubble destruction (UTMD) is a new type of gene delivery technology. MiR-21-5p was highly expressed in a variety of cancers. In this paper, miR-21-5p inhibitor was transfected into lung cancer cells by UTMD to observe its role in lung cancer.
StarBase was used to analyze the miR-21-5p expression in lung cancer patients and its relationship with the prognosis of the patients. MiR-21-5p expression in lung cancer tissues or cell lines was determined by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Effects of gradient concentration (0, 5, 10, 20, 30%) of SonoVue or gradient mechanical index (MI) (0, 0.5, 1, 1.5, 2 W/cm) on the cell viability were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The targeting relationship between miR-21-5p and B-cell translocation gene 2 (BTG2) was predicted by TargetScan and confirmed by dual-luciferase reporter assay, while the expressions of the two genes were determined by qRT-PCR. Through liposome transfection or UTMD transfection, the effects of miR-21-5p/BTG2 on the biological behaviors of lung cancer cells, the size of xenograft tumors and the expressions of ki67 and miR-21-5p were measured by qRT-PCR, western blot, cell function experiments and immunohistochemistry, respectively.
MiR-21-5p expression was upregulated in lung cancer, which was associated with a poor prognosis. The optimal ultrasound conditions were 10% SonoVue concentration and 1 W/cm. UTMD transfection exerted a stronger effect than liposome transfection. MiR-21-5p promoted cell viability, proliferation and migration yet suppressed apoptosis by targeting BTG2. MiR-21-5p inhibitor reduced the size and volume of xenograft tumor and the expressions of ki67 and miR-21-5p in xenograft tumor tissues.
UTMD-mediated miR-21-5p inhibitor can more effectively suppress the development of lung cancer.
超声靶向微泡破坏（UTMD）是一种新型的基因传递技术。miR-21-5p 在多种癌症中高表达。本文通过 UTMD 将 miR-21-5p 抑制剂转染入肺癌细胞，观察其在肺癌中的作用。
利用 StarBase 分析肺癌患者 miR-21-5p 的表达及其与患者预后的关系。采用实时定量逆转录聚合酶链反应（qRT-PCR）检测肺癌组织或细胞系中 miR-21-5p 的表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测不同浓度（0、5、10、20、30%）声诺维或不同机械指数（MI）（0、0.5、1、1.5、2 W/cm）对细胞活力的影响。通过 TargetScan 预测 miR-21-5p 与 B 细胞易位基因 2（BTG2）的靶向关系，并通过双荧光素酶报告基因实验进行验证，同时采用 qRT-PCR 检测这两个基因的表达。通过脂质体转染或 UTMD 转染，采用 qRT-PCR、western blot、细胞功能实验和免疫组织化学法分别检测 miR-21-5p/BTG2 对肺癌细胞生物学行为、异种移植瘤大小以及 ki67 和 miR-21-5p 表达的影响。
miR-21-5p 在肺癌中表达上调，与预后不良相关。最佳超声条件为 10%声诺维浓度和 1 W/cm。UTMD 转染比脂质体转染效果更强。miR-21-5p 通过靶向 BTG2 促进细胞活力、增殖和迁移，抑制细胞凋亡。miR-21-5p 抑制剂降低了异种移植瘤的大小和体积，以及异种移植瘤组织中 ki67 和 miR-21-5p 的表达。
UTMD 介导的 miR-21-5p 抑制剂能更有效地抑制肺癌的发展。
