长链非编码RNA（lncRNA）在肿瘤恶化中发挥重要作用。前期工作中，我们发现上调hnRNPAB能促进肝癌侵袭转移，且与Kap1蛋白互作和lncRNA2012表达调控密切相关；进一步发现过表达lncRNA2012可抑制肝癌细胞运动和侵袭。本课题通过共转染hnRNPAB和lncRNA2012过表达质粒或shRNA，进行体内外功能分析，明确lncRNA2012介导的hnRNPAB在肝癌侵袭转移中的作用；应用染色质免疫沉淀、荧光素报告基因分析hnRNPAB表达对lncRNA2012转录调控的影响；通过免疫共沉淀和免疫印迹明确hnRNP AB和Kap1的互作，观察shRNA-Kap1干预前后肝癌细胞内hnRNPAB亚细胞定位和转录活性的改变及其对调控lncRNA2012表达和肝癌侵袭转移的影响，同时分析三者在肝癌组织中的表达及与肝癌转移复发间的关系。本研究为探讨肝癌治疗中新的干预靶点提供实验依据。

Long chain non encoding RNA (lncRNA),a class of non encoding RNA with transcription length of more than 200 nucleotides, plays an important role in tumor progression. In our previous study, we found that over-expression of heterogeneous nuclear ribonucleoprotein AB (HNRNP AB) in hepatocellular carcinoma (HCC) cells promoted cell invasion in vitro and lung metastasis in vivo; this effect may be associated with its interaction with Kap1 and its transcriptional regulation of lncRNA2012 expression. Further study shows that over-expression of lncRNA-2012 can inhibit the migration and invasion of HCC, suggesting that lncRNA-2012 can promote HCC metastasis, which might be regulated by HNRNP AB. In this study, through short hairpin RNA (shRNA) interference or up-regulation of hnRNP AB and lncRNA2012 expression in HCC cell lines with various metastatic potentials, we will assess their roles in tumor invasion and in vivo metastasis and determine the role of lncRNA2012 in HNRNP AB-enhanced HCC invasion and metastasis. Furthermore, chromatin immunoprecipitation (ChIP) and luciferase assays will be used to analyze whether hnRNP AB is involved in the transcriptional regulation of lncRNA2012 expression in HCC cells. Subsequently, western blotting and co-immunoprecipitation will be used to determine the interaction between hnRNP AB and Kap1. Through shRNA-mediated down-regulation of Kap1 in HCC cells, we will determine the changes on cellular location of hnRNP AB and its transcriptional activity. We will also assess its roles on regulation of lncRNA2012 expression induced by hnRNP AB and determine its roles on tumor invasion and in vivo metastasis. The relationship between the expression of these genes and the invasive phenotype of HCC cells will be studied. We will determine whether these genes could be important independent factors in determining clinical outcomes of HCC. This study will provide a biological foundation for the discovery of new potential therapeutic targets for HCC.