Qu Qiang, Chu Xiangdong, Wang Peng
Department of Orthopedics, Affiliated Hospital of Shaanxi University of Chinese Medicine, Xianyang 712000, China.
Cell Biol Int. 2017 Mar;41(3):287-295. doi: 10.1002/cbin.10723. Epub 2017 Jan 11.
MiR-195-5p has been shown to play an essential role in human cancer progression. Nevertheless, the biological role of miR-195-5p in osteosarcoma development remains unclear. In this study, real-time PCR was performed to examine the miR-195-5p expression in human osteosarcoma cell lines. CCK-8 assay and Transwell assay were carried out to measure the effect of miR-195-5p on cell proliferation and invasion. Luciferase reporter assay was used to identify the targets of miR-195-5p. The results showed that miR-195-5p was significantly downregulated in osteosarcoma cells. Forced expression of miR-195-5p significantly inhibited cell proliferation, suppressed cell migration and invasion, compared with wild-type and control-transfected osteosarcoma cells. Luciferase reporter assay revealed that miR-195-5p binds to the 3'-untranslated region (UTR) of Naked cuticle homolog 1 (NKD1), indicating that NKD1 was a novel target of miR-195-5p. NKD1 mRNA and protein levels were reduced after overexpression of miR-195-5p. Moreover, silencing of NKD1 significantly inhibited the proliferation and invasion of osteosarcoma cells. Accordingly, our results support a tumor suppressor role of miR-195-5p in osteosarcoma through inhibiting NKD1, and it may be a promising therapeutic target for osteosarcoma.
已证明miR-195-5p在人类癌症进展中起重要作用。然而，miR-195-5p在骨肉瘤发生中的生物学作用仍不清楚。在本研究中，采用实时定量PCR检测人骨肉瘤细胞系中miR-195-5p的表达。进行CCK-8检测和Transwell检测以测量miR-195-5p对细胞增殖和侵袭的影响。使用荧光素酶报告基因检测来鉴定miR-195-5p的靶标。结果显示，miR-195-5p在骨肉瘤细胞中显著下调。与野生型和对照转染的骨肉瘤细胞相比，miR-195-5p的强制表达显著抑制细胞增殖，抑制细胞迁移和侵袭。荧光素酶报告基因检测显示，miR-195-5p与裸角质层同源物1（NKD1）的3'-非翻译区（UTR）结合，表明NKD1是miR-195-5p的新靶标。miR-195-5p过表达后，NKD1的mRNA和蛋白水平降低。此外，沉默NKD1可显著抑制骨肉瘤细胞的增殖和侵袭。因此，我们的结果支持miR-195-5p通过抑制NKD1在骨肉瘤中发挥肿瘤抑制作用，并且它可能是骨肉瘤的一个有前景的治疗靶点。
