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新型微小RNA-9500通过靶向Akt1调控人肺癌细胞的增殖和迁移。

The novel miR-9500 regulates the proliferation and migration of human lung cancer cells by targeting Akt1.

作者信息

Yoo J K, Jung H Y, Lee J M, Yi H, Oh S-H, Ko H Y, Yoo H, Kim H-R, Song H, Kim S, Kim J K

机构信息

Department of Pharmacy, College of Pharmacy, CHA University, Yatap-dong, Bundang-gu, Seongnam-si, Gyeonggi-do, Republic of Korea.

Department of Neurology, CHA Bundang Medical Center, CHA University, Seongnam-si, Gyeonggi-do, Republic of Korea.

出版信息

Cell Death Differ. 2014 Jul;21(7):1150-9. doi: 10.1038/cdd.2014.33. Epub 2014 Mar 21.

Abstract

MicroRNAs have crucial roles in lung cancer cell development. They regulate cell growth, proliferation and migration by mediating the expression of tumor suppressor genes and oncogenes. We identified and characterized the novel miR-9500 in human lung cancer cells. The miR-9500 forms a stem-loop structure and is conserved in other mammals. The expression levels of miR-9500 were reduced in lung cancer cells and lung cancer tissues compared with normal tissues, as verified by TaqMan miRNA assays. It was confirmed that the putative target gene, Akt1, was directly suppressed by miR-9500, as demonstrated by a luciferase reporter assay. The miR-9500 significantly repressed the protein expression levels of Akt1, as demonstrated via western blot, but did not affect the corresponding mRNA levels. Akt1 has an important role in lung carcinogenesis, and depletion of Akt1 has been shown to have antiproliferative and anti-migratory effects in previous studies. In the current study, the overexpression of miR-9500 inhibited cell proliferation and the expression of cell cycle-related proteins. Likewise, the overexpression of miR-9500 impeded cell migration in human lung cancer cells. In an in vivo assay, miR-9500 significantly suppressed Fluc expression compared with NC and ASO-miR-9500, suggesting that cell proliferation was inhibited in nude mice. Likewise, miR-9500 repressed tumorigenesis and metastasis by targeting Akt1. These data indicate that miR-9500 might be applicable for lung cancer therapy.

摘要

微小RNA在肺癌细胞发展过程中发挥着关键作用。它们通过介导肿瘤抑制基因和癌基因的表达来调节细胞生长、增殖和迁移。我们在人肺癌细胞中鉴定并表征了新型的miR - 9500。miR - 9500形成茎环结构,并且在其他哺乳动物中保守。经TaqMan miRNA检测验证,与正常组织相比,miR - 9500在肺癌细胞和肺癌组织中的表达水平降低。荧光素酶报告基因检测表明,miR - 9500可直接抑制假定的靶基因Akt1。蛋白质印迹法显示,miR - 9500显著抑制Akt1的蛋白质表达水平,但不影响相应的mRNA水平。Akt1在肺癌发生过程中起重要作用,先前的研究表明,Akt1缺失具有抗增殖和抗迁移作用。在本研究中,miR - 9500过表达抑制细胞增殖及细胞周期相关蛋白的表达。同样,miR - 9500过表达阻碍人肺癌细胞的迁移。在体内实验中,与阴性对照(NC)和反义寡核苷酸miR - 9500(ASO - miR - 9500)相比,miR - 9500显著抑制荧光素酶(Fluc)表达,表明裸鼠体内细胞增殖受到抑制。同样,miR - 9500通过靶向Akt1抑制肿瘤发生和转移。这些数据表明,miR - 9500可能适用于肺癌治疗。

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