The functional analysis and clinical application of the metastasis-associated gene TI-241.

我们对具有不同转移潜力的黑色素瘤亚群进行了鉴别筛选，并分离出它们之间差异表达的几个基因。这些基因中的大多数已经被报道过，但有些还没有。在这些研究中，TI227与正式报道的基因没有同源性，并且在具有不同转移潜力的结肠癌细胞亚群中表达不同。该基因在结肠癌细胞的高实验转移亚系中表达较多。我们分析了它的序列，并分离出了人类对应的TI227H。通过对TI227和TI227H的同源性研究，确定了最可能的ORF。我们进一步分离了含有TI227的基因组DNA，并分析了其5'上游序列。TI227有望用于诊断转移潜力和预后。另一个基因TI241与LRF1和ATP3的转录因子具有同源性。转染低转移亚克隆增强了其转移潜能，其在转移灶中的表达升高。在使用手术切除标本的研究中，其表达在血管侵犯的情况下表达更多，并且其作为预后预测因子的重要性被期望。同样从反义寡核苷酸的研究中，TI241被证明与细胞粘附和侵袭有关，并被认为与体内生长有关。从裸鼠实验来看，它抑制了皮下生长，延长了它们的生存期，这为其应用于基因治疗开辟了可能性。以上结果均已发表或投稿至英文期刊。

We performed differential screening between melanoma sublines of dirrerent metastatic potentials and isolated several genes that were differentially expressed between them. Most of these genes were already reported, but some were not. In those, TI227 showed no homology with formaly reported genes and differentally expressed in the sublines of colon cancer cells with different metastatic potential. This gene was expressed more in the highly experimentally metastatic subline of colon cancer cells. We analyzed its sequence and isolated its human counterpart TI227H. From the homology research between TI227 and TI227H, the most possible ORF was determined. We further isolated genomic DNA containing TI227 and analysed its 5' upstream sequence. TI227 was expected to be used in the diagnosis of metastatic potential and prognosis. Another gene TI241 showed homology with transcription factors of LRF1 and ATP3. Its transfection to the low metastatic subclone strengthened its metastatic potential and its expression in the metastatic foci was elevated. In the study using surgically excised specimens, its expression was expressed more in the cases with vascular invasions and its importance as a prognosis predictor was expected. Also from the study with the antisense oligonucleotides, TI241 was shown to relate cell adhesion and invasion and was thought to relate in vivo growth. From the exp6riments using nude mice, it surpressed subcutaneus growth and lengthened their survival period, which opened a possibility of its application to the gene therapy. Above results were already published or submitted to the English journals.