Ding Hui, Li Liangpeng, Gu Biao, Ni Yaojun, Chen Sheng
Department of Thoracic Surgery, Huai'an First People's Hospital, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huai'an, Jiangsu 223300, P.R. China.
Department of Thoracic Surgery, Nanjing First Hospital, Nanjing Medical University, P.R. China.
Exp Ther Med. 2021 Apr;21(4):358. doi: 10.3892/etm.2021.9789. Epub 2021 Feb 13.
Νon-small cell lung cancer (NSCLC) is the most frequently diagnosed type of cancer, and the most prevalent cause of cancer-associated mortality. The present study aimed to investigate whether microRNA (miR)-564 influences NSCLC progression by regulating NSCLC cell growth and migration, via targeting plexin A4. Therefore, the expression levels of miR-564 and plexin A4 were evaluated in NSCLC specimens or cells using reverse transcription-quantitative PCR. Furthermore, colony formation and Cell Counting Kit-8 assays were performed to determine the proliferative ability of NSCLC cells. The cell migration capacity was assessed using a Transwell assay. In addition, to examine the binding ability of miR-564 on the plexin A4 3'-untranslated region (3'UTR), a dual-luciferase reporter assay was performed. A mouse xenograft model was established to evaluate the effect of miR-564 knockdown on tumor growth , whereas the expression of plexin A4 and Ki67 in NSCLC tissues was detected using immunohistochemistry. Notably, miR-564 was downregulated in both NSCLC cell lines and tissues, while its overexpression, following transfection with miR-564 mimics, attenuated the proliferation and proliferation, migration and invasion of NSCLC cells. By contrast, silencing of miR-564 using a miR-564 inhibitor promoted NSCLC cell proliferation, migration and invasion. The luciferase assay revealed that miR-564 directly targeted the plexin A4 3'UTR in A549 and H460 cells. Additionally, the overexpression of plexin A4 rescued the effect of miR-564 on NSCLC cell proliferation, migration and invasion abilities. Further studies demonstrated that miR-564 knockdown promoted NSCLC growth, while miR-564 overexpression resulted in the opposite effect in nude mice. Overall, the results of the present study revealed that miR-564 promotes the proliferation and migration of NSCLC cells, both and , via targeting plexin A4. Therefore, miR-564 may be considered as a possible therapeutic target for NSCLC.
非小细胞肺癌（NSCLC）是最常被诊断出的癌症类型，也是癌症相关死亡的最常见原因。本研究旨在探讨微小RNA（miR）-564是否通过靶向丛状蛋白A4调节NSCLC细胞生长和迁移来影响NSCLC进展。因此，使用逆转录定量PCR评估NSCLC标本或细胞中miR-564和丛状蛋白A4的表达水平。此外，进行集落形成和细胞计数试剂盒-8测定以确定NSCLC细胞的增殖能力。使用Transwell测定评估细胞迁移能力。此外，为了检测miR-564与丛状蛋白A4 3'-非翻译区（3'UTR）的结合能力，进行了双荧光素酶报告基因测定。建立小鼠异种移植模型以评估miR-564敲低对肿瘤生长的影响，而使用免疫组织化学检测NSCLC组织中丛状蛋白A4和Ki67的表达。值得注意的是，miR-564在NSCLC细胞系和组织中均下调，而在用miR-564模拟物转染后其过表达减弱了NSCLC细胞的增殖、迁移和侵袭。相比之下，使用miR-564抑制剂沉默miR-564可促进NSCLC细胞增殖、迁移和侵袭。荧光素酶测定显示miR-564直接靶向A549和H460细胞中的丛状蛋白A4 3'UTR。此外，丛状蛋白A4的过表达挽救了miR-564对NSCLC细胞增殖、迁移和侵袭能力的影响。进一步的研究表明，miR-564敲低促进了NSCLC生长，而miR-564过表达在裸鼠中产生相反的效果。总体而言，本研究结果表明，miR-564通过靶向丛状蛋白A4促进NSCLC细胞的增殖和迁移。因此，miR-564可能被视为NSCLC的一个可能治疗靶点。
