肝细胞癌骨转移：稳定的小鼠肿瘤模型系统的建立及分子机制研究

肝细胞癌是最常见的恶性肿瘤之一，进展期骨转移多见且呈逐年上升趋势，给病人造成严重痛苦。由于缺乏模拟人肝细胞癌骨转移过程的模型系统，基于模型的肝癌骨转移分子机制研究未见报道。本研究拟解决以下问题：一.利用肝癌细胞系或临床肝癌标本，行肝脏原位种植或左心室注射，建立自发性或实验性肝癌骨转移动物模型。二.行"肝－骨"及"左心室－骨"连续体内筛选三代以上，获得具有特异性骨转移潜能的肝癌细胞系，并分析其与亲代细胞在增殖、侵袭、成瘤等表型差异。三.利用具有特异性骨转移潜能的肝癌细胞系，与其亲代细胞系行基因表达谱芯片及miRNA芯片检测并分析差异，寻找与肝癌骨转移相关的候选基因及miRNA。四.临床收集人肝癌及骨转移癌标本，验证上述候选基因/miRNA的表达,确定肝癌骨转移相关基因及miRNA。五.开展体内/外实验，探索肝癌骨转移相关基因及miRNA的作用机制，为肝癌骨转移的治疗提供新的思路和理论依据。

Hepatocellular carcinoma (HCC) is one of the commonest malignant tumours. An increasingly high occurrence of bone metastases in HCC patients highlights the importance of fundamental research on HCC bone metastasis, which have been in poor situation for long due to lack of suitable model system. In this research program, we are supposed to solve the following problems: 1. Establishment of experimental and spontaneous animal model of HCC bone metastasis by intracardiac injection or tumour xenograft orthotopic transplantation respectively. 2. Establishment of HCC cell lines with unique potential to form bone metastasis (BM cells) by means of successive "liver to bone"or"heart to bone"in vivo selection (more than 3 passages). 3. mRNA-based expression microarray and miRNA microarray to analyze differences between BM cells and parental cells, finding candidates for HCC bone metastasis-related genes and miRNAs. 4. Collection of patients' tissue samples of primary HCC and bone metastasis, corroboration of HCC bone metastasis-related genes and miRNAs in tissue samples. 5. Comprehensive oncological in vitro/in vivo assays to explore the mechanisms of HCC bone metastasis-related genes and miRNAs, and to elucidate its potential value in clinic.