血清长链非编码RNA作为宫颈癌新型分子标记物的筛查及功能研究

A growing body of research suggests that Long non-coding RNA (LncRNA) is closely related to the occurrence and development of tumor and can be used as new molecular markers of tumor diagnosis and treatment, but the research in cervical cancer is rare. Recently, we confirmed that specific LncRNA can be detected in serum of cervical cancer patients. Subsequently, we independently developed a tumor-related LncRNA expression profile microarray. We used this microarray to investigate samples from cervical cancer patients (n=10) and healthy women (n=10). Preliminary confirmed specific LncRNA expression profile exists in the serum of cervical cancer patients. This project will test 120 cases of serum from cervical cancer patients and 120 cases of serum from healthy female volunteers using the tumor related LncRNA expression profile microarray to identify the LncRNA expression profile and screen out the specific LncRNA with differentially expressed(LncRNA-X) in cervical cancer. Furthermore, we will determine the expression level of LncRNA-X using real time qRT-PCR, explore its clinical significance combined with clinical data analysis, reveal its molecular biology function by overexpressing or knocking down LncRNA-X expression in vitro and in vivo. The molecular targets of LncRNA-X will be identified using RNA Pull Down and mass spectrometry technology to clarify the underlying molecular mechanisms. The implementation of this project will establish a novel screening approach for cervical cancer tumor markers and has important clinical implications for the diagnosis and treatment of cervical cancer.

越来越多的研究表明，长链非编码RNA(LncRNA)与肿瘤的发生发展密切相关，可作为肿瘤诊断和治疗的新型分子标记物，但其在宫颈癌中的研究甚少。近日，我们证实在血清中可检测到宫颈癌特异性LncRNA，并自主研发出一款肿瘤相关LncRNA表达谱基因芯片，利用这款芯片试检测了10例宫颈癌和10例健康女性血清，初步证实宫颈癌患者血清中存在特异性的LncRNA表达谱。本课题拟用上述基因芯片检测120例宫颈癌和120例健康女性血清，鉴定出宫颈癌血清LncRNA表达谱，筛查出宫颈癌特异性差异表达LncRNA，结合临床病理资料分析明确其临床意义；对有重要临床意义的LncRNA用基因过表达及基因沉默的方法对其进行功能研究，揭示其分子生物学功能；用RNA Pull Down及质谱分析鉴定其作用的靶蛋白，探索其发挥功能的作用机制。本项目的实施将开辟宫颈癌肿瘤标记物筛查的新方向，对宫颈癌的临床诊治有重要的意义。

长链非编码RNA(LncRNA)与肿瘤的发生发展密切相关，可作为肿瘤诊断和治疗的新型分子标记物，但其在宫颈癌中的研究甚少。本项目在前期研究的基础上，建立了LncRNA探针微阵列，筛查出11个宫颈癌相关LncRNA。验证出XLOC_012799在宫颈癌中表达明显上调，并且其高表达与淋巴结转移及患者的不良预后明显相关；为明确XLOC_012799生物学功能，通过RACE实验获得了其全长序列；功能探索研究发现XLOC_012799可促进宫颈癌细胞侵袭及转移；机制研究发现XLOC_012799在宫颈癌细胞中可与TBL1XR1竞争性结合miR-454-3p，抑制 miR-454-3p与TBL1XR1mRNA 3'UTR结合，使TBL1XR1蛋白翻译表达增加，从而促进细胞的转移。本项目的实施对理解和阐明宫颈癌的转移具有重要的科学意义，对宫颈癌的分子靶向治疗具有重要的临床意义。