Lerner Mikael, Harada Masako, Lovén Jakob, Castro Juan, Davis Zadie, Oscier David, Henriksson Marie, Sangfelt Olle, Grandér Dan, Corcoran Martin M
Department of Oncology-Pathology, Cancercentrum Karolinska, Karolinska Institutet R8:03, 171 76 Stockholm, Sweden.
Exp Cell Res. 2009 Oct 15;315(17):2941-52. doi: 10.1016/j.yexcr.2009.07.001. Epub 2009 Jul 8.
The microRNAs miR-15a and miR-16-1 are downregulated in multiple tumor types and are frequently deleted in chronic lymphocytic leukemia (CLL), myeloma and mantle cell lymphoma. Despite their abundance in most cells the transcriptional regulation of miR-15a/16-1 remains unclear. Here we demonstrate that the putative tumor suppressor DLEU2 acts as a host gene of these microRNAs. Mature miR-15a/miR-16-1 are produced in a Drosha-dependent process from DLEU2 and binding of the Myc oncoprotein to two alterative DLEU2 promoters represses both the host gene transcript and levels of mature miR-15a/miR-16-1. In line with a functional role for DLEU2 in the expression of the microRNAs, the miR-15a/miR-16-1 locus is retained in four CLL cases that delete both promoters of this gene and expression analysis indicates that this leads to functional loss of mature miR-15a/16-1. We additionally show that DLEU2 negatively regulates the G1 Cyclins E1 and D1 through miR-15a/miR-16-1 and provide evidence that these oncoproteins are subject to miR-15a/miR-16-1-mediated repression under normal conditions. We also demonstrate that DLEU2 overexpression blocks cellular proliferation and inhibits the colony-forming ability of tumor cell lines in a miR-15a/miR-16-1-dependent way. Together the data illuminate how inactivation of DLEU2 promotes cell proliferation and tumor progression through functional loss of miR-15a/miR-16-1.
微小RNA miR-15a和miR-16-1在多种肿瘤类型中表达下调,且在慢性淋巴细胞白血病(CLL)、骨髓瘤和套细胞淋巴瘤中常发生缺失。尽管它们在大多数细胞中含量丰富,但miR-15a/16-1的转录调控仍不清楚。在此,我们证明了假定的肿瘤抑制因子DLEU2是这些微小RNA的宿主基因。成熟的miR-15a/miR-16-1通过Drosha依赖的过程从DLEU2产生,Myc癌蛋白与两个交替的DLEU2启动子结合会抑制宿主基因转录本以及成熟miR-15a/miR-16-1的水平。与DLEU2在微小RNA表达中的功能作用一致,在4例CLL病例中,该基因的两个启动子均缺失,但miR-15a/miR-16-1基因座得以保留,表达分析表明这导致成熟miR-15a/16-1功能丧失。我们还表明,DLEU2通过miR-15a/miR-16-1负调控G1期细胞周期蛋白E1和D1,并提供证据表明在正常条件下这些癌蛋白受到miR-15a/miR-16-1介导的抑制。我们还证明,DLEU2过表达以miR-15a/miR-16-1依赖的方式阻断细胞增殖并抑制肿瘤细胞系的集落形成能力。这些数据共同阐明了DLEU2的失活如何通过miR-15a/miR-16-1的功能丧失促进细胞增殖和肿瘤进展。