Sánchez-Beato M, Camacho F I, Martínez-Montero J C, Sáez A I, Villuendas R, Sánchez-Verde L, García J F, Piris M A
Department of Genetics and Department of Pathology, Virgen de la Salud Hospital, Toledo, Spain.
Blood. 1999 Jul 15;94(2):765-72.
p27 cyclin-dependent kinase inhibitor downregulation is essential for transition to the S phase of the cell cycle. Thus, proliferating cells in reactive lymphoid tissue show no detectable p27 expression. Nevertheless, anomalous high p27 expression has been shown to be present in a group of aggressive B-cell lymphomas with high proliferation index and adverse clinical outcome. This suggests that abnormally accumulated p27 protein has been rendered functionally inactive. We analyzed the causes of this anomalous presence of p27 in a group of aggressive B-cell lymphomas, including 54 cases of diffuse large B-cell lymphomas and 20 Burkitt's lymphomas. We simultaneously studied them for p27, cyclin D3, cyclin D2, cyclin D1, and cyclin E expression, because it has been stated that high levels of expression of cyclin D1 or E lead to increased p27 levels in some cell types. A statistically significant association between p27 and cyclin D3 expression was found for the group as a whole. Additionally, when dividing the cases according to the level of expression of cyclin D3 by reactive germinal centers, it was observed that cases with stronger cyclin D3 expression also show higher p27 expression. The relationship between both proteins was also shown at a subcellular level by laser confocal studies, showing that in cases with high expression of both proteins there was a marked colocalization. Additional evidence in favor of p27 sequestration by cyclin D3 was provided by coimmunoprecipitation studies in a Burkitt's cell line (Raji) showing the existence of cyclin D3/p27 complexes and the absence of CDK2/p27 complexes. These results could support the hypothesis that there are cyclin D3/p27 complexes in a subset of aggressive B-cell lymphomas in which p27 lacks the inhibitory activity found when it is bound to cyclin E/CDK2 complexes. This interaction between both proteins could lead to an abnormal nuclear accumulation, detectable by immunohistochemical techniques.
p27细胞周期蛋白依赖性激酶抑制剂的下调对于细胞周期进入S期至关重要。因此,反应性淋巴组织中的增殖细胞未显示可检测到的p27表达。然而,在一组具有高增殖指数和不良临床结局的侵袭性B细胞淋巴瘤中,已显示存在异常高的p27表达。这表明异常积累的p27蛋白已丧失功能活性。我们分析了一组侵袭性B细胞淋巴瘤中p27异常存在的原因,包括54例弥漫性大B细胞淋巴瘤和20例伯基特淋巴瘤。我们同时研究了它们的p27、细胞周期蛋白D3、细胞周期蛋白D2、细胞周期蛋白D1和细胞周期蛋白E表达,因为有研究表明在某些细胞类型中,细胞周期蛋白D1或E的高表达会导致p27水平升高。对于整个研究组,发现p27与细胞周期蛋白D3表达之间存在统计学上的显著关联。此外,根据反应性生发中心细胞周期蛋白D3的表达水平对病例进行划分时,观察到细胞周期蛋白D3表达较强的病例也显示出较高的p27表达。激光共聚焦研究还在亚细胞水平上显示了这两种蛋白之间的关系,表明在两种蛋白高表达的病例中存在明显的共定位。在伯基特细胞系(Raji)中进行的免疫共沉淀研究提供了支持细胞周期蛋白D3隔离p27的额外证据,显示存在细胞周期蛋白D3/p27复合物,而不存在CDK2/p27复合物。这些结果可能支持这样的假设,即在一部分侵袭性B细胞淋巴瘤中存在细胞周期蛋白D3/p27复合物,其中p27缺乏与细胞周期蛋白E/CDK2复合物结合时所具有的抑制活性。这两种蛋白之间的这种相互作用可能导致异常的核内积累,可通过免疫组织化学技术检测到。