Prediction of in vitro response to interferon-alpha and acquisition of its susceptibility by gene and peptide transduction system

通过基因和肽转导系统预测干扰素-α的体外反应并获取其敏感性

基本信息

批准号：
18591739
负责人：
SHIMAZUI Toru
金额：
$ 2.53万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

项目摘要

It is important to investigate the interferon-alpha (IFN-a) sensitivity-related gene and regulation of susceptibility to IFN-α in renal cell carcinoma (RCC). First, we have identified seven candidate genes, which are associated with IFN-a-response and then established prediction formula using four genes, I.e. ADFP, MITF, MTUS1, and TNNT1, in RCC cell lines. Validation of this prediction model was performed in other RCC cell lines, including primary culture cells from patients with RCC. We next focused MITF gene, which individually correlated with IFN-α-sensitivity, to analyze relationship between its expression and IFN-α-response status by gene transfection. MITF transfectant demonstrated susceptibility in IFN-α-resistant line as compared with wild type or mock transfectant. Because MITF is one off transcription factor, which can bind to p16 and HIF-1α, it is suggested that MITF may be associated with development and progression of RCC acceleration of cell cycle. Thus, we evaluated expression of p16 and anti-tumor effect of biological active p16 peptide (p16-MIS) by peptide transduction system with Wr-T transporter peptide. All RCC cell lines used are absent for p16 protein, even in the IFN-α-resistant RCC MITF-transfectant. P16-MIS successfully transferred into cellular cytoplasm and nucleus of RCC cells by concentration dependent manner. Interestingly, IFN-α-susceptibility was acquired at 2.5 to 100 times intensity in IFN-α-resistant RCC cell line. These results suggested that IFN-α-response related gene, MITF and its related cell cycle regulator, p16 could be molecular targets in therapeutic strategy of RCC.

研究肾细胞癌（RCC）中干扰素-α（IFN-a）敏感性相关基因及其对 IFN-α 敏感性的调节非常重要。首先，我们鉴定了七个与 IFN-a 相关的候选基因。 -响应，然后使用四个基因，即 ADFP、MITF、MTUS1 和 TNNT1，在 RCC 细胞系中建立预测公式，并在其他 RCC 细胞系中进行了该预测模型的验证。我们接下来关注与 IFN-α 敏感性相关的 MITF 基因，通过基因转染证明其表达与 IFN-α 反应状态之间的关系。由于 MITF 是一种可以与 p16 和 HIF-1α 结合的转录因子，因此表明 MITF 可能与发育和进展有关。因此，我们通过Wr-T转运蛋白肽转导系统评估了p16的表达和生物活性p16肽（p16-MIS）的抗肿瘤作用。所有使用的RCC细胞系均不存在p16蛋白。，甚至在 IFN-α 抗性 RCC MITF 转染子中，P16-MIS 也以浓度依赖性方式成功转移到 RCC 细胞的细胞质和细胞核中。在 IFN-α 耐药的 RCC 细胞系中获得了 2.5 至 100 倍强度的 IFN-α 敏感性。这些结果表明 IFN-α 反应相关基因 MITF 及其相关细胞周期调节因子 p16 可以作为治疗的分子靶点。农村信用社战略。