miR-365b调控肝癌侵袭转移的机制研究

MicroRNAs (miRNAs) are a class of small, single strand, non-coding RNA molecules. They regulate the expression of target genes through post-transcriptionally regulating and function in various cellular processes. The previous study found that several miRNAs located in chromosome fragile sites and differentially expressed in hepatocellular carcinoma (HCC); however, the role and function of most of these miRNAs in HCC were still unknown. In this study, we first found that miR-365b which is amplified and upregulated in HCC could regulate liver cancer cell migration and invasion. Furthermore, these observations were confirmed by gain-of-function and loss-of-function assays and the experiment in vivo would be performed. Additionally, it is planned to search the potential target gene for miR-365b in HCC by integrating bioinformatics prediction, microarray analysis, protein mass spectrometry and luciferase assays. Still, we intend to detect the expression of the target gene in HCC through immunohistochemical staining assays and examine its clinical significance. In addition, the function of the target gene and the signal pathway is to be explored. In conclusion, this study is to comprehensively investigate the expression, function and molecular mechanisms of miR-365b in HCC. It is hopeful that the findings would provide novel potential targets for HCC therapy and prognosis.

MicroRNA是一类小分子单链非编码RNA，具有转录后调控基因表达的功能。前期研究发现一批与肝癌染色体脆性位点相关且癌与癌旁表达显著差异的miRNA分子，但它们在肝癌中的功能尚不完全清楚。本研究发现在肝癌组织中DNA拷贝数增加、表达显著上调的miR-365b，可参与调节肝癌细胞的迁移和侵袭。随后利用多种肝癌细胞系，并通过上调/下调miR-365b从正反两方面进行功能分析，接着拟通过体内实验以明确miR-365b对肝癌侵袭转移的作用。进一步拟结合基因芯片技术与生物信息学软件、蛋白质谱以及荧光素酶活性检测，寻找miR-365b 的直接靶基因；免疫组化检测该基因在肝癌中的表达并分析其临床意义；同时进行相关功能实验以观察其在肝癌中的作用及探索其下游通路。 综上，本论文拟探讨肝癌相关miR-365b及其靶基因在肝癌中的表达及功能，并对其相关分子作用机制进行研究，希望为肝癌的治疗和预后提供新的靶标。

MicroRNA（miRNAs）是一类进化上高度保守的小分子非编码RNA，主要在转录后负调控基因表达，且与肿瘤的发生发展密切相关。前期研究发现miR-365b与肝癌染色体脆性位点相关，在肝癌组织中DNA拷贝数增加、表达显著上调，但它在肝癌中的功能尚不完全清楚。本研究发现miR-365b可参与调节肝癌细胞的迁移和侵袭。首先检测其在多种肝癌细胞系的表达情况，并通过上调/下调miR-365b进行功能分析，明确了miR-365b对肝癌细胞的促迁移、侵袭作用。接着通过mRNA芯片与生物信息学软件筛出一批候选靶基因，进一步通过real-time PCR、western 实验和荧光素酶活性检测进行验证，发现SGTB（small glutamine-rich tetratricopeptide repeat (TPR)-containing, beta）可能为miR-365b 的直接靶基因；该基因在肝癌中的表达及其临床意义有待通过免疫组化检测进一步分析；同时其在肝癌中的作用也需相关功能实验进一步明确。 .综上，本课题探讨了miR-365b在肝癌中的表达及功能，并对其相关分子机制进行了研究，初步发现了其直接靶基因SGTB，尚待补充实验进一步明确，希望能为肝癌的治疗和预后提供新的靶标。