长链非编码RNA-AK002107在肝细胞癌侵袭转移中的作用及其分子机制

Epithelial-mesenchymal transition (EMT) is an important mechanism for tumor invasion and metastasis. Our previous study found that the expression of lncRNA-AK0020107 was up-regulated and was closely associated with malignant pathological features of hepatocellular carcinoma (HCC). It could promote the invasion and migration of HCC cells with concomitant EMT phenomenon. Then we found.that it was co-expressed with chemokine ligand 21 (CCL21) in HCC tissues. Down-regulation of lncRNA-AK002107 by siRNA inhibited the expression of CCL21 in HCC cell lines. Thus, we speculate that lncRNA-AK002107 may promote HCC invasion and metastasis by induction of EMT via regulating CCL21/CCR7. Firstly, to investigate the influence of lncRNA-AK0020107 on HCC EMT, invasion and migration, meanwhile detect the expression of CCL21 and CCR7. Then, through animal study, to determine lncRNA-AK002107 promotes HCC invasion and metastasis by induction of EMT through regulating CCL21/CCR7. Finally, to analyze the clinical significance of lncRNA-AK002107 by detecting its expression in HCC tissues. This study aims to elucidate the molecular mechanism how lncRNA-AK0020107 promote HCC invasion and metastasis and provide scientific basis for lncRNA-AK002107 acting as a novel target of targeted therapy for HCC.

上皮-间质转化（EMT）是肿瘤侵袭转移的重要机制。我们前期研究发现lncRNA-AK0020107在肝细胞癌组织中高表达，与肝癌的恶性病理特征密切相关，体外预实验证实其能促进肝癌细胞侵袭转移且伴有显著的EMT现象。继而分析发现在肝癌组织中其与趋化因子配体21（CCL21）共表达，下调其表达后，CCL21表达也下调。由此，我们假设：lncRNA-AK002107通过调控CCL21/CCR7诱导EMT促进肝细胞癌侵袭转移。本项目拟首先观察该基因对肝细胞癌EMT和侵袭转移等生物学行为的影响，并检测该过程中CCL21、CCR7的表达水平变化；其次采用动物实验证实该基因通过调控CCL21/CCR7诱导EMT促进肝细胞癌侵袭转移，最后在临床标本中分析其表达的临床意义。本课题旨在阐明lncRNA-AK002107促进肝细胞癌侵袭转移的分子机制，为该基因作为肝细胞癌治疗新靶点提供科学依据。

长链非编码RNA的异常表达与人类肿瘤密切相关，本项目的和主要内容是研究lncRNA AK002107在肝细胞癌进展中的作用和潜在机制。结果显示lncRNA AK002107在肝细胞癌中高表达，其高表达与甲胎蛋白、肿瘤大小、大血管侵犯和微血管侵犯密切相关。高表达lncRNA AK002107患者的无瘤生存率和总体生存率明显低于低表达患者。体外功能实验证实抑制肝癌细胞lncRNA AK002107的表达，细胞的增殖、克隆形成和侵袭能力明显减弱。体内皮下移植瘤结果显示抑制lncRNA AK002107表达后，肝癌细胞体内生长受抑制。通过生信分析并荧光素酶报告基因验证lncRNA AK002107直接结合miR-140-5p并能够抑制miR-140-5p的表达。细胞实验证实lncRNA AK002107在肝细胞癌起致癌作用是通过调节miR-140-5p的表达。最后机制分析发现lncRNA AK002107负性调节miR-140-5p表达，进而靶向调控TGFBR1表达诱导肝细胞上皮-间质转化促进肝癌进展。本研究证实lncRNA AK002107/miR-140-5p/TGFBR1/EMT调控网络在肝细胞癌发生和进展中起重要作用，有望发展成为肝癌新颖诊断和治疗方法的有价值靶标。