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

我们对不同转移潜能的黑色素瘤亚系进行了差异筛选，并分离出了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了，但也有一些没有报道。其中，TI227基因与已报道的基因没有同源性，在不同转移潜能的结肠癌细胞亚系中有不同的表达。该基因在高实验转移性的结肠癌细胞亚系中表达更多。我们分析了它的序列，并分离出了它的人类同源基因TI227H。从TI227和TI227H的同源性研究中，确定了最可能的ORF。我们进一步提取了含有TI227的基因组DNA，并对其5‘上游序列进行了分析。TI227有望用于转移潜能的诊断和预后判断。另一个基因TI241与转录因子LRF1和ATP3同源。对低转移亚克隆的转染增强了其转移潜能，在转移灶中的表达增强。在使用手术切除标本的研究中，它的表达在有血管侵袭的病例中表达得更多，其作为预后预测指标的重要性被预期。与反义寡核苷酸的研究也表明，TI241与细胞黏附和侵袭有关，并被认为与体内生长有关。裸鼠实验表明，它能抑制皮下组织生长，延长其存活时间，为其应用于基因治疗提供了可能。上述研究成果已在英文期刊上发表或提交。

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.