新基因C6orf223在肾癌发生发展中的分子机制研究

Renal cell carcinoma accounts for adult tumor 2-3%, its mechanism is still not clarified.Our prior transcriptome analysis revealed that the expression of open reading frame C6orf223 was higher in clear cell renal cell carcinoma (ccRCC) tissues compared to that in matched normal renal tissues, which was confirmed by quantitative real-time PCR (RT-PCR） and immunohistochemical staining (IHC);C6orf223 was low expression or no expression in a panel of renal cancer cell lines, and Hypoxia can induce C6orf223 gene expression in renal cancer cell lines; ectopic expression of C6orf223 can promote proliferation of renal cancer cells,which suggested that C6orf223 is hypoxia-inducible gene and is associated with renal caner tumorigenesis. However, there is no report on C6orf223 in MEDLINE database. In order to confirm the hypothesis, we will first detect C6orf223 expression by RT-PCR, Western blot and IHC, and determinate the association between its expression and clinicopathological parameters in large ccRCC cancer samples. Then, we will investigate whether C6orf223 is a novel direct target of HIF-1α or HIF-2α by Luciferase reporter assay, and chromatin immunoprecipitation assay. At last, we will investigate the biological function and signaling pathway of C6orf223 in ccRCC by the lentiviral expression system, nude mice xenograft tumor model, and transcriptome analysis to clarify the roles of C6orf223 in renal cell carcinoma and its molecular mechanisms, and provide new clues to early diagnosis and gene therapy for renal cell carcinoma.

肾癌约占成人肿瘤的2-3%，其发生机制仍未阐明。本课题组通过对10例肾透明细胞癌的转录组分析，筛选出一个在癌组织中表达显著高于癌旁正常组织的开放阅读框C6orf223基因。荧光定量PCR及Western blot检测证实C6orf223基因在肾癌组织中表达上调；低氧可诱导肾癌细胞株中C6orf223表达上调；过表达C6orf223显著促进肾癌细胞增殖，上述结果提示C6orf223参与了肾癌的发生发展，可能是个低氧诱导的新候选癌基因。本课题拟采用RT-PCR、Western blot、免疫组化、荧光素酶报告系统、染色质免疫共沉淀、慢病毒载体和裸鼠模型等方法研究肾癌组织、细胞及裸鼠移植瘤中C6orf223基因的表达特征，低氧诱导C6orf223基因促进肾癌发生的信号通路及其分子调控机制。上述研究有望进一步揭示肾癌发生的分子机制，并为肾癌的早期诊断及靶标治疗提供科学依据。

肾癌是泌尿系统最常见的肿瘤之一，约占成人肿瘤的2-3%，但其发生机制仍未阐明。本课题组通过对10例肾透明细胞癌的转录组分析，筛选出一个在癌组织中表达显著高于癌旁正常组织的开放阅读框C6orf223基因，并通过qPCR、Western blot和免疫组织化学染色方法证实在肾癌组织和细胞系中表达显著上调；低氧可促进C6orf223在肾癌细胞中的表达，进而通过调节下游EMT相关分子表达，促进肾癌细胞的侵袭和转移；另外miR-532-3p与C6orf223表达成负相关，miR-532-3p可直接靶向调控C6orf223，进而影响肾癌细胞的浸润和迁移。上述研究有望进一步揭示肾癌发生的分子机制，并为肾癌的早期诊断及靶标治疗提供科学依据。