Liu Bao-Yan, Lu Yan-Qin, Han Feng, Wang Yong, Mo Xin-Kai, Han Jin-Xiang
Shandong Medical Biotechnological Center, School of Medicine and Life Science, Shandong Academy of Medical Sciences, University of Jinan, Jinan, Shandong 250062, P.R. China.
Key Laboratory for Rare Disease Research of Shandong, Shandong Medical Biotechnological Center, Shandong Academy of Medical Sciences, Jinan, Shandong 250062, P.R. China.
Oncol Lett. 2017 Jan;13(1):111-118. doi: 10.3892/ol.2016.5426. Epub 2016 Nov 23.
The present study aimed to investigate the effects of overexpression of interferon-induced transmembrane protein 5 (IFITM5) and IFITM5 c.-14C>T mutation on osteogenic differentiation, and the proliferation, migration and invasion of SaOS2 cells. SaOS2 cells were transfected with plasmids containing wild type IFITM5 (W) or IFITM5 containing the c.-14C>T mutation (MU). The mRNA and protein expression levels of IFITM5 in SaOS2 cells were respectively detected by reverse transcription quantitative polymerase chain reaction and western blotting. The proliferative, migratory and invasive ability of SaOS2 cells was also examined. In addition, the expression levels of osteogenic differentiation markers alkaline phosphatase (ALP), osteocalcin (OCN) and runt-related transcription factor 2 (Runx2) were detected. Mineralized nodules were detected by Alizarin Red S staining and were quantified by measuring absorbance. The mRNA and protein expression levels of IFITM5 were high in cells transfected with IFITM5 and IFITM5 c.-14C>T mutation, and were higher in cells transfected with IFITM5 c.-14C>T mutation. There was no difference in proliferation between the control group (C) and the W and MU groups. However, overexpression of IFITM5 and IFITM5 c.-14C>T mutation increased apoptotic rate, decreased invasive capacity, increased the expression of ALP, OCN and Runx2, and increased the number of mineralized nodules following osteogenic induction. In addition, compared with C and W groups, cells transfected with IFITM5 c.-14C>T mutation exhibited decreased migratory ability. In conclusion, overexpression of IFITM5 and IFITM5 c.-14C>T mutation promotes tumor cell apoptosis, inhibits tumor invasion and promotes osteogenic differentiation. These findings may provide a theoretical basis for the development of a novel treatment method that targets IFITM5, and provides a platform for the potential treatment of human osteosarcoma.
本研究旨在探讨干扰素诱导跨膜蛋白5（IFITM5）过表达及IFITM5基因c.-14C>T突变对成骨分化以及SaOS2细胞增殖、迁移和侵袭的影响。将含有野生型IFITM5（W）或含有c.-14C>T突变的IFITM5（MU）的质粒转染至SaOS2细胞。通过逆转录定量聚合酶链反应和蛋白质免疫印迹法分别检测SaOS2细胞中IFITM5的mRNA和蛋白表达水平。同时检测SaOS2细胞的增殖、迁移和侵袭能力。此外，检测成骨分化标志物碱性磷酸酶（ALP）、骨钙素（OCN）和 runt相关转录因子2（Runx2）的表达水平。通过茜素红S染色检测矿化结节，并通过测量吸光度进行定量分析。在转染IFITM5和IFITM5基因c.-14C>T突变的细胞中，IFITM5的mRNA和蛋白表达水平较高，且在转染IFITM5基因c.-14C>T突变的细胞中更高。对照组（C）与W组和MU组之间的增殖无差异。然而，IFITM5过表达和IFITM5基因c.-14C>T突变可增加凋亡率，降低侵袭能力，增加ALP、OCN和Runx2的表达，并在成骨诱导后增加矿化结节的数量。此外，与C组和W组相比，转染IFITM5基因c.-14C>T突变的细胞迁移能力降低。总之，IFITM5过表达和IFITM5基因c.-14C>T突变可促进肿瘤细胞凋亡，抑制肿瘤侵袭并促进成骨分化。这些发现可能为开发以IFITM5为靶点的新型治疗方法提供理论依据，并为人类骨肉瘤的潜在治疗提供平台。
