Knockdown of FBXO39 inhibits proliferation and promotes apoptosis of human osteosarcoma U-2OS cells.
Knockdown of FBXO39 inhibits proliferation and promotes apoptosis of human osteosarcoma U-2OS cells.
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DOI：
10.3892/ol.2018.8876
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发表时间：
2018-06
期刊：
影响因子：
2.9
通讯作者：
Jianrong Zheng;W. You;Chuanxi Zheng;Peng Wan;Jinquan Chen;Xiaochun Jiang;Zhixiang Zhu;Zhixiong Zhang;Anqi Gong;Wei Li;Ji-Hong Tan;T. Ji;Wei Guo;Shiquan Zhang
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文献类型：
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作者：
Jianrong Zheng;W. You;Chuanxi Zheng;Peng Wan;Jinquan Chen;Xiaochun Jiang;Zhixiang Zhu;Zhixiong Zhang;Anqi Gong;Wei Li;Ji-Hong Tan;T. Ji;Wei Guo;Shiquan Zhang
F-box蛋白是Skp-cullin-F-box复合物（一种E3泛素连接酶）的重要组成部分，通过泛素蛋白酶体系统参与细胞周期和免疫应答。F-box蛋白39（FBXO 39）属于F-box家族，已被报道与癌症的发生和进展相关。本研究旨在探讨FBXO 39在体外骨肉瘤细胞增殖和凋亡中的作用。结果显示U-2OS细胞中FBXO 39的表达高于HOS和SaOS-2细胞。因此，使用慢病毒介导的短发夹RNA（shRNA）转染进行FBXO39的敲低，以验证FBXO39在U-2OS细胞中的作用。使用Western印迹和RT-qPCR分析通过分析FBXO39的表达水平来确认感染效率。使用基于Celigo的细胞计数和MTT测定，证明与对照相比，FBXO 39敲低显著降低了细胞增殖速率。Caspase 3/7活性测定和荧光激活细胞分选证实了FBXO 39敲低后U-2OS细胞中的凋亡诱导。结论：FBXO39基因敲减可显著抑制U-2OS细胞增殖并促进其凋亡。因此，FBXO 39可能在OS进展中发挥重要作用。
F-box proteins are essential components of the Skp-cullin-F-box complex (a type of E3 ubiquitin ligase), and participate in cell cycle and immune responses through the ubiquitin proteasome system. F-box protein 39 (FBXO39) belongs to the F-box family, which has been reported to be associated with cancer oncogenesis and progression. The present study aimed to investigate the role of FBXO39 in osteosarcoma (OS) cell proliferation and apoptosis in vitro. It was demonstrated that U-2OS cells exhibited high expression of FBXO39 compared with HOS and SaOS-2 osteosarcoma cells. Thus, knockdown of FBXO39 was performed using lentivirus-mediated short hairpin RNA (shRNA) transfection to validate the effect of FBXO39 in U-2OS cells. Western blotting and RT-qPCR analysis were used to confirm the efficiency of infection by analyzing the expression level of FBXO39. Using Celigo-based cell counting and MTT assays, it was demonstrated that FBXO39 knockdown significantly reduced the rate of cell proliferation compared with control. Caspase 3/7 activity assays and fluorescence-activated cell sorting confirmed the induction of apoptosis in U-2OS cells following FBXO39 knockdown. In conclusion, it was demonstrated that FBXO39 knockdown may significantly inhibit proliferation and promote apoptosis of U-2OS cells. Thus, FBXO39 may serve an important role in OS progression.