Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.
Current address: Department of Pediatrics, University of Rome, Tor Vergata, Rome, Italy.
J Cell Mol Med. 2009 Aug;13(8B):1666-1678. doi: 10.1111/j.1582-4934.2008.00501.x.
The ubiquitin C-terminal hydrolase-L1 (UCH-L1) is a deubiquitinating enzyme that catalyses the hydrolysis of polyubiquitin precursors and small ubiquitin adducts. UCH-L1 has been detected in a variety of malignant and metastatic tumours but its biological function in these cells is unknown. We have previously shown that UCH-L1 is highly expressed in Burkitt's lymphoma (BL) and is up-regulated upon infection of B lymphocytes with Epstein-Barr virus (EBV). Here we show that knockdown of UCH-L1 by RNAi inhibits the proliferation of BL cells in suspension and semisolid agar and activates strong LFA-1-dependent homotypic adhesion. Induction of cell adhesion correlated with cation-induced binding to ICAM-1, clustering of LFA-1 into lipid rafts and constitutive activation of the Rap1 and Rac1 GTPases. Expression of a catalytically active UCH-L1 promoted the proliferation of a UCH-L1-negative EBV transformed lymphoblastoid cell line (LCL) and inhibited cell adhesion, whereas a catalytic mutant had no effect, confirming the requirement of UCH-L1 enzymatic activity for the regulation of these phenotypes. Our results identify UCH-L1 as a new player in the signalling pathways that promote the proliferation and invasive capacity of malignant B cells.
泛素 C 端水解酶-L1(UCH-L1)是一种去泛素化酶,可催化多泛素前体和小泛素缀合物的水解。UCH-L1 已在多种恶性和转移性肿瘤中被检测到,但在这些细胞中的生物学功能尚不清楚。我们之前曾表明,UCH-L1 在 Burkitt 淋巴瘤(BL)中高度表达,并在 B 淋巴细胞感染 EBV 时被上调。在这里,我们表明,通过 RNAi 敲低 UCH-L1 可抑制悬浮和半固体琼脂中 BL 细胞的增殖,并激活强烈的 LFA-1 依赖性同源黏附。细胞黏附的诱导与阳离子诱导与 ICAM-1 的结合、LFA-1 聚集到脂筏中和 Rap1 和 Rac1 GTPase 的组成性激活相关。表达具有催化活性的 UCH-L1 促进了 UCH-L1 阴性 EBV 转化的淋巴母细胞系(LCL)的增殖,并抑制了细胞黏附,而催化突变体没有作用,证实了 UCH-L1 酶活性对这些表型的调节的必要性。我们的研究结果表明,UCH-L1 是促进恶性 B 细胞增殖和侵袭能力的信号通路中的一个新成员。