Yang Ming, Liu Shi Chang, Hao Ding Jun, Yan Liang, Liu Zhong Kai, Yin Xin Hua
Department of Spine Surgery, Hong Hui Hospital, Xi'an Jiaotong University College of Medicine Xi'an, Shaanxi, China.
Am J Cancer Res. 2022 Mar 15;12(3):1264-1281. eCollection 2022.
Chordoma is a rare bone tumor, and the recurrence rate of chordoma is high, the treatment is difficult, and the prognosis is poor. Therefore, it is of great significance to find key target genes for the treatment of chordoma. Microarray was used to analyze the significant gene associated with chordoma. Western blot and RT-PCR were used to detect protein and mRNA expression levels of RP11-867G2.8 and FUT4. Fluorescence in situ hybridization (FISH) assay was used to locate the position of RP11-867G2.8 in chordoma cells. MTT assay, colony formation assay, transwell assay and Xenograft Mouse Model were used to clarify the function of RP11-867G2.8 and FUT4. RNA pull-down, RNA immunoprecipitation, RNA stability assay and polysome profiling analysis were used to clarify the relationship between RP11-867G2.8 and FUT4. We found that RP11-867G2.8 is highly expressed in chordoma tissues and cells, and RP11-867G2.8 overexpression promotes the malignant biological behavior of chordoma cells. RP11-867G2.8 overexpression alters the expression pattern of genes modulating signaling pathway. FUT4 is accumulated in chordoma tissues, and RP11-867G2.8 is antisense RNA of FUT4. RP11-867G2.8 can bind to FUT4 mRNA, increasing FUT4 mRNA stability and facilitating translation of FUT4. RP11-867G2.8 binds to EIF4B and PABPC1, which increases the translation of FUT4. Further studies found that FUT4 silence counteracts the effect of RP11-867G2.8 and . Our results suggest that RP11-867G2.8 promotes the development and progression of chordoma by up-regulating the expression of FUT4.
脊索瘤是一种罕见的骨肿瘤，脊索瘤的复发率高，治疗困难，预后较差。因此，寻找治疗脊索瘤的关键靶基因具有重要意义。利用基因芯片分析与脊索瘤相关的显著基因。采用蛋白质免疫印迹法（Western blot）和逆转录-聚合酶链反应（RT-PCR）检测RP11-867G2.8和岩藻糖基转移酶4（FUT4）的蛋白质和mRNA表达水平。采用荧光原位杂交（FISH）试验确定RP11-867G2.8在脊索瘤细胞中的位置。采用MTT法、集落形成试验、Transwell试验和异种移植小鼠模型阐明RP11-867G2.8和FUT4的功能。采用RNA下拉试验、RNA免疫沉淀试验、RNA稳定性试验和多核糖体谱分析阐明RP11-867G2.8与FUT4之间的关系。我们发现RP11-867G2.8在脊索瘤组织和细胞中高表达，RP11-867G2.8过表达促进脊索瘤细胞的恶性生物学行为。RP11-867G2.8过表达改变了调节信号通路的基因表达模式。FUT4在脊索瘤组织中蓄积，且RP11-867G2.8是FUT4的反义RNA。RP11-867G2.8可与FUT4 mRNA结合，增加FUT4 mRNA稳定性并促进FUT4的翻译。RP11-867G2.8与真核翻译起始因子4B（EIF4B）和多聚腺苷酸结合蛋白1（PABPC1）结合，从而增加FUT4的翻译。进一步研究发现，FUT4沉默可抵消RP11-867G2.8的作用。我们的结果表明，RP11-867G2.8通过上调FUT4的表达促进脊索瘤的发生和发展。
