miR-330-5p抑制肝细胞癌复发转移作用机制的研究

The long-term prognosis of HCC remains unsatisfactory, mainly due to the high recurrence and metastasis rate of HCC. In our previous study, miR-330-5p was frequently up-regulated in HCC. Moreover, knock-down the expression of miR-330-5p can effectively inhibit the HCCLM3 cells invasion and metastasis. Therefore, we speculated that miR-330-5p could promote tumor cells invasion and metastasis in HCC. In this study, we plan to use lentiviral vector to knock down the expression of miR-330-5p and then use wound healing assay, transwell assay and HCC nude mouse model to validate the role of miR-330-5p in recurrence and metastasis of HCC. Then we will adopt bioinformatics, protein chip, luciferase reporter assay to screen the target genes and find out the molecular mechanisms. Finally, we will build the glycosylation of polyethylenimine (PEI) nanoparticles mediated by miR-330-5p targeted therapy system to find out the feasibility of HCC therapy by inhibiting the expression of miR-330-5p. In conclusion, the aim of this study is to clarify the role of miR-330-5p in recurrence and metastasis of human hepatocellular carcinoma, which would provide us a wider prospective on HCC intervetion/prevention and treatment.

肝癌的术后复发转移是制约其长期生存水平提高的关键因素。我们前期研究显示miR-330-5p在肝癌组织中呈显著高表达，敲除miR-330-5p的表达能有效地抑制肝癌细胞系HCCLM3的侵袭转移能力。故我们推测miR-330-5p很可能具有较强的促进肝癌侵袭转移的作用。为此，我们拟采用慢病毒介导的敲除技术、划痕实验、Transwell及肝癌裸鼠成瘤模型等进一步验证miR-330-5p在肝癌侵袭转移中的作用；继而采用生物信息学、蛋白芯片、荧光素酶报告基因筛选其靶基因并进行系统的体外、体内研究，以期阐明其作用的分子机制；最后拟构建糖基化修饰的聚乙烯亚胺（PEI）纳米颗粒介导的miR-330-5p靶向治疗系统，探讨其体内系统干预对肝癌侵袭转移能力的抑制作用。研究miR-330-5p及其靶基因的作用和机制能够让我们更好地理解肝癌发展过程中的分子机制并为研究肝癌的防治新措施提供了一个更好的前景。

MicroRNAs在肿瘤的生长和增殖中发挥重要作用，这些是导致肝癌发生侵袭转移的重要原因。我们的前期研究发现miR-330-5p在肝癌组织中往往呈高表达。但是，它的作用机制尚不明确。在本研究中我们进一步确认了miR-330-5p在肝癌组织及细胞系中呈高表达，且与肝癌的肿瘤大小、数目、有无包膜以及TNM分期、肝癌患者生存预后密切相关。而且通过体内体外实验证实过表达miR-330-5p可促进肝癌的生长和增殖，而敲除miR-330-5p则抑制其生长增殖。接着我们通过双荧光素梅报告基因系统进一步证实在肝癌中SPRY2是miR-330-5p的靶基因。最后我们通过一系列体内体外实验证实在肝癌中miR-330-5p抑制SPRY2的表达来调控 MAPK/ERK信号通路活性从而发挥调控肝癌进展的作用。我们的研究结果将为肝癌的靶向治疗提供一个非常有应用价值的靶点及为肝癌患者生存预后提供一个有价值的预后标志物。