一个大鼠肝再生相关的新lncRNA（rLALR1）调节肝细胞增殖的作用及机理研究

In recent years, studies have shown that lncRNAs have important biological functions, involving in cell proliferation and apoptosis. Our laboratory have revealed that a new lncRNA-rLALR1 has an important role in rat hepatocyte proliferation and liver regeneration (LR) by high-throughput sequencing, gene expression profile chip detection, bioinformatics analysis and preliminary experiment, and its target gene have been detected as SIK1 preliminarily, but its mechanism is unclear. To investigate the mechanism of rLALR1 regulating hepatocyte proliferation and LR, we will study the distribution of rLALR1 in hepatoma cells, primary hepatocyte that cultured in vitro and regenerating liver qualitatively and quantitatively at first, reveal the regulation mechanism of rLALR1 and target gene by RNA pull down、ChIRP-seq, and detect the gene and protein expression changes and its influence on cell proliferation, cell cycle, etc. in vitro and in vivo after rLALR1 was over-expressed or knocked down. The function channels, ways and molecular mechanism of rLALR1 in regulating hepatocyte proliferation will be analyzed during rat LR. This study will set a basis for us to understand the mechanism of liver regeneration and liver disease and establish the treatment and prevention method of liver disease.

近几年研究发现，lncRNA具有重要的生物学功能，其中一些参与了细胞增殖与凋亡的调节。本实验室通过lncRNA高通量测序、基因表达谱芯片检测、生物信息学分析和初步实验发现，一个新的lncRNA—rLALR1在大鼠肝细胞增殖和肝再生中起重要作用，并初步确定其靶基因是SIK1，但其作用机理尚不清楚。为探讨rLALR1调控肝细胞增殖和肝再生的机理，本项目拟先在肝癌细胞系、原代肝细胞及再生肝中对rLALR1进行定性、定位和定量研究，并用RNA pull down、ChIRP-seq等方法验证其调控下游靶基因的机制，然后分别从体外和体内两个水平检测rLALR1或其靶基因过表达或沉默后相关基因和蛋白的表达变化以及对细胞增殖、细胞周期等的影响，从而解析rLALR1的作用途径和方式，阐明大鼠肝再生中rLALR1调控肝细胞增殖的分子机理，为了解肝再生和肝脏疾病发生的机制、建立治疗和预防肝病方法奠定基础。

肝再生调控的分子机制复杂而有序，非编码RNA也在其中发挥了重要的作用。近几年研究发现，lncRNA具有重要的生物学功能，其中一些参与了细胞增殖与凋亡的调节。本项目通过lncRNA高通量测序、基因表达谱芯片检测、生物信息学分析和初步实验发现，一个新的lncRNA—rLALR1（rat lncRNA associated with liver regeneration 1）在大鼠肝细胞增殖和肝再生中起重要作用，并初步确定其靶基因是SIK1，但其作用机理尚不清楚。为探讨rLALR1调控肝细胞增殖和肝再生的机理，本项目在大鼠肝癌细胞系、大鼠正常肝细胞系和再生肝中对rLALR1进行定性和定量研究，并用RNA pull down、ChIRP-seq等方法验证其调控下游靶基因的机制，然后分别从体外和体内两个水平检测rLALR1或其靶基因过表达或沉默后相关基因和蛋白的表达变化以及对细胞增殖、细胞周期等的影响，揭示rLALR1能正向调节下游靶基因SIK1表达促进肝再生中肝细胞增殖，推测大鼠肝再生中rLALR1调控肝细胞增殖的分子机理，但验证rLALR1如何正向调节SIK1的表达遇到了一定困难，还在进一步优化实验条件。此外，分析了肝再生整个进程中lncRNA相关数据，以及lncRNA与mRNA，miRNA的互作情况。本研究为了解肝再生和肝脏疾病发生的机制、建立治疗和预防肝病方法奠定基础。