lncRNA-NONHSAT024778竞争性结合miR-1290调控Robo1促进脊索瘤增殖、侵袭的作用及机制研究

Chordoma is a malignant bone tumor known to arise from the embryonic remnants of the notochord. The invasiveness of chordoma is strong, recurrences and metastasis usually occurred, and the prognosis is poor. Long non-coding RNAs (lncRNA), which may act as competing endogenous RNAs (ceRNAs), play a critical role in the pathological process of tumors. However, their biological role and underlying molecular mechanism in chordoma remains largely unknow. Our previous study has found that lncRNA NONHSAT024778 high-regulation is inversely correlated with the upregulation of miR-1290 in chordoma, which were also predicted by bioinfomatics. We also found that NONHSAT024778 could supress the expression of miR-1290 in vitro experiments. Furthermore, the results of bioinformatics analysis suggests that NONHSAT024778 may function as a ceRNA to regulate the expression of the downstream target gene Robo1 by sponging miR-1290. Therefore, we propose the hypothesis: NONHSAT024778 functions as a ceRNA to regulate Robo1 by sponging miR-1290 and promote the proliferation and invasion of chordoma. To investigate the effect and mechanism of NONHSAT024778 in the regulation of miR-1290 in the progression of chordoma, we utilize a series of experiments in vitro and in vivo, including examining chordoma tissue samples, constructing NONHSAT024778 overexpression/ deletion cell model, and establishment of the nude mice model, cell transfection, RNA Binding Protein Immunoprecipitation (RIP) and luciferase reporter assays. This study has important significance to clarify the proliferation and invasion mechanism of chordoma and will provide us new ideas and experimental basis to early diagnosis and targeted therapy.

脊索瘤侵袭性强，复发、转移率高，预后差。研究表明lncRNA可作为ceRNA参与肿瘤的增殖、侵袭，而其在脊索瘤中的作用尚未阐明。我们的前期研究发现lncRNA-NONHSAT024778在脊索瘤中高表达且与miR-1290的表达负相关；体外实验发现NONHSAT024778可抑制miR-1290的表达；生物信息学结果显示NONHSAT024778可吸附miR-1290并作用于靶基因Robo1。因此，我们提出假设：NONHSAT024778可竞争性结合miR-1290调控Robo1促进脊索瘤增殖、侵袭。本研究通过脊索瘤组织、构建NONHSAT024778过/缺失表达脊索瘤细胞模型及裸鼠模型，采用细胞转染、RNA免疫共沉淀、荧光报告基因等技术，多层面、系统的研究脊索瘤中NONHSAT024778的生物作用及其作为ceRNA的分子调控机制，以期为脊索瘤早期诊断和靶向治疗提供新的思路和实验基础。

项目背景：脊索瘤侵袭性强，复发、转移率高，预后差。研究表明lncRNA可作为ceRNA参与肿瘤的增殖、侵袭，而其在脊索瘤中的作用尚未阐明。我们的前期研究发现 lncRNA-NONHSAT024778在脊索瘤中高表达；生物信息学结果显示NONHSAT024778可吸附miRNA并作用于靶基因。因此，我们提出假设：NONHSAT024778可竞争性结合miRNA-1290调控靶基因促进脊索瘤增殖、侵袭。本研究通过脊索瘤组织、构建NONHSAT024778过/缺失表达脊索瘤细胞模型及裸鼠模型，采用细胞转染、RNA免疫共沉淀荧光报告基因等技术，多层面、系统的研究脊索瘤中NONHSAT024778的生物作用及其作为ceRNA的分子调控机制，以期为脊索瘤早期诊断和靶向治疗提供新的思路和实验基础。主要研究内容包括：（1）研究脊索瘤中NONHSAT024778的表达情况。（2）研究NONHSAT024778对脊索瘤细胞增殖、侵袭能力的影响。（3）研究NONHSAT024778竞争性结合miR-1290促进脊索瘤增殖、侵袭的机制。（4）研究脊索瘤裸鼠模型中NONHSAT024778对脊索瘤增殖、侵袭的影响。研究结果：脊索瘤组织中NONHSAT024778表达增加与miR-1290水平降低相关。NONHSAT024778基因敲除可抑制脊索瘤细胞的增殖和侵袭。miR-1290的表达恢复了NONHSAT024778的致癌功能。生物信息学分析表明，NONHSAT024778作为ceRNA，通过在脊索瘤细胞中分泌miR-1290来调节Robo1，从而促进脊索瘤细胞恶性进展。体内实验结果证实了NONHSAT024778敲除激活miR-1290抑制癌基因Robo1的抗肿瘤作用。NONHSAT024778在脊索瘤组织中大量过度表达，而miR-1290在脊索瘤组织中减少。科学意义：本研究为脊索瘤的早期诊断和靶向治疗提供新的思路和实验依据。如可开发出适合临床应用的脊索瘤抗靶点药物，可直接降低医疗费用，并通过降低患者的复发率及致死率，改善患者身心健康，提高生活质量，使更多的劳动力能重返社会，对脊索瘤的预防干预及有效的诊断和治疗具有广阔的应用前景，并将获得深远的社会经济效益。