结直肠癌肝转移相关miRNA对肝转移的预测及其对结直肠癌细胞系的生物学影响

项目名称： 结直肠癌肝转移相关miRNA对肝转移的预测及其对结直肠癌细胞系的生物学影响

项目编号： No.30872467

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 顾晋

作者单位： 北京市肿瘤防治研究所

项目金额： 31万元

中文摘要： 利用miRNA芯片技术筛选无肝转移的CRC原发灶和伴有同时性肝转移的CRC原发灶两组间差异表达的miRNAs。发现两组miRNA表达谱有显著差异，其中有7个miRNA表达显著不同。采用实时定量PCR（LNA探针法）和LNA标记寡核苷酸探针原位杂交方法验证了差异表达miRNAs在CRC组织中的表达情况。结果表明miR-181a和miR-320在伴有同时性肝转移的CRC原发灶（PSLM）和异时性肝转移的CRC原发灶(PMLM) 组的表达要高于无肝转移的CRC原发灶(PNM)组（P<0.05）。生存分析表明miR-181a与患者的预后呈负相关。构建携带miR-181a前体的慢病毒载体并转染入结直肠癌细胞系HT29细胞系，观察基因转染对肿瘤细胞生长、迁移、体外浸润以及免疫缺陷鼠皮下移植瘤生长的影响。结果表明，慢病毒介导的miR-181a基因转染可促进HT29细胞在体外的生长、迁移和浸润能力，同时还可促进免疫缺陷鼠皮下移植瘤的生长。因此miR-181a与CRC的异时性肝转移密切相关，可能在CRC肝转移过程中发挥重要作用，有望成为预测肝转移的分子标志物。。

中文关键词： miRNA;肠癌；肝转移

英文摘要： We first isolated and compared the miRNA expression profile from CRC and CRC liver matastasis(CLM) tumor tissue by microarray. The pattern of miRNA expression in CRC was markedly different from that of CLM. Out of total miRNAs, 7 (miR-181a, 152,320,let-7d, miR-15b, 20b,223) were found to be markedly deregulated in CLM when compared with CRC. miRNAs expression was validated by realtime PCR (Locked-nucleic acid probe method) and LNA-in situ hybridization, the levels of miR-181a and 320 were much higher in the CRL than in the matched normal tissues and CRC. The level of miR-181a in metachronous LM is much higher than in synchronous LM. We found that the expression level of miR-181a was also correlated significantly with overall survival in CRC patients. We constructed a lentivirus vector expressing the human mir-181a and a green fluorescent protein (GFP), transfected the miR-181a overexpressing vector into HT29 cells and established a stable cell line.We found that miR-181a enhanced the proliferation, migration and invasion of HT29 cell in vitro. To further explore the role of miR-181a in tumor growth in vivo, HT29 cells stably expressing miR-181a were transplanted into the nude mice. We found that miR-181a enhanced tumor growth in vivo. Taken together, miR-181a might be a potential predictive factor or therapeutic target for colorectal liver matastasis.

英文关键词： miRNA;colorectal cancer; liver matastasis