Molecular analysis of Cyclin D1 in the development of mantle cell lymphoma

套细胞淋巴瘤（Mantle cell lymphoma，MCL）是近年来发现的难治性淋巴瘤之一，其中t（11;14）引起的Cyclin D1过表达是其特异性特征。然而，cyclin D1在淋巴瘤中的具体作用尚未完全阐明。为了检验它，我们最初计划通过引入PTD（达特）融合的抗细胞周期蛋白D1的单链抗体来抑制细胞周期蛋白D1的功能，所述单链抗体可以从产生抗细胞周期蛋白D1的单克隆抗体的杂交瘤（5D4）中分离。由于使用达特寡肽作为PTD的初步研究表明对白血病/淋巴瘤细胞（包括MCL细胞系）的引入效率相当低，因此我们需要开发更有效的用于将肽/蛋白质递送至造血细胞的系统。我们还重新考虑了引入p16 INK 4a功能肽可能比使用抗cyclin D1的ScFv更优选用于抑制cyclin D1的功能，因为它是Cdk 4/Cyclin D1复合物的特异性抑制剂。将p16功能肽 ...更多信息 在肿瘤淋巴细胞中，我们着重研究了肽转运蛋白系统，并试图开发出比以前的系统效率更高的转运蛋白。因此，我们成功地产生了新的肽转运蛋白，其具有扩展的肽对接结构域和细胞渗透结构域，其用于增强货物肽到白血病/淋巴瘤细胞中的转导效率。研究表明，这种新型转运蛋白能够将肽靶向进入MCL细胞，其靶向能力是之前肽递送系统的20-30倍，这可以将货物肽的量减少到之前系统的1/50-1/100。根据这一结果，我们将使用p16功能肽的该系统应用于高度侵袭性的白血病/淋巴瘤细胞系（MCL、Burkitt淋巴瘤、NK/T细胞淋巴瘤、CML的母细胞性改变），如果生长抑制发生或不发生，则结果显示在所有这些白血病/淋巴瘤细胞中观察到显著的抑制（最大约85%）。此外，p16肽引入的新的转运蛋白在小鼠模型中引起体内伯基特肿瘤的显着的生长迟缓，这意味着该系统的效用，在体内淋巴瘤。我们完成了数据的总结，现在正在向《医学科学杂志》投稿。部分相关数据发表在Eur. J. Immunol.第33卷，第1 -11页，2003年，Kondo等人，Less

Mantle cell lymphoma (MCL) has been recently recognized as one of the intractable lymphomas, and overexpression of Cyclin D1 caused by t(11;14) is identified as specific feature in them. However, concrete function of cyclin D1 in the lymphoma has not been fully disclosed yet. To examine it, we initially planed to suppress the function of Cyclin D1 by introducing PTD(TAT)-fused ScFv against cyclin D l, which can be isolated from anti-cyclin D1 MoAb-producing hybridoma (5D4). Because the pilot study using TAT oligopeptide as PTD demonstrated quite low introduction efficiency to leukemia/lymphoma cells including MCL cell lines, we needed to develop more efficient system for peptide/protein-delivery to hematopoietic cells. We also reconsidered that introducing p16 INK4a functional peptide could be more preferable to using ScFv against cyclin D1 for inhibition of cyclin D1's function because it is a specific inhibitor of the Cdk4/Cyclin D1 complexes. To introduce the p16 functional peptide … More into neoplastic lymphoid cells, we focused on peptide transporter system and attemped to develop the transporter with higher efficiency than that of previous system. Consequently, we succeeded in generating the novel peptide transporter that has both the expanded peptide-docking domain and the cell-permeable domain which serves to enhance the transduction efficiency of a cargo peptide into leukemia/lymphoma cells. The novel transporter was revealed to enable to target the peptide into MCL cells 20-30 times greater than the previous peptide delivery systems, which could reduce the amount of a cargo peptide to 1/50-1/100 of that in the previous systems. Following this result, we applied this system using the p16 functional peptide to highly aggressive leukemia/lymphoma cell lines (MCL, Burkitt's lymphomas, NK/T-cell lymphomas, blastic change of CML) if the growth inhibition took place or not.The result showed that remarkable suppression was observed in all of these leukemia/lymphoma cells (maximum 〜85%). Moreover, the p16 peptide introduced by the novel transporter caused a dramatic growth retardation of in vivo Burkitt's tumors in mouse models, which meant the utility of this system to in vivo lymphomas. We finished summarizing the data and are now submitting the manuscript to the journal of medical science. Partially related data were published in Eur. J. Immunol. Vol.33, p1-11, 2003 by Kondo et al. Less