A novel human heparanase splice variant, T5, endowed with protumorigenic characteristics
A novel human heparanase splice variant, T5, endowed with protumorigenic characteristics
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DOI：
10.1096/fj.09-147074
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发表时间：
2010-04-01
期刊：
影响因子：
4.8
通讯作者：
Vlodavsky, Israel
中科院分区：
文献类型：
--
作者：
Barash, Uri;Cohen-Kaplan, Victoria;Vlodavsky, Israel
乙酰肝素酶是一种哺乳动物内切β-D-葡萄糖醛酸酶，可切割硫酸乙酰肝素侧链，这种活性与肿瘤细胞播散密切相关。目前的研究旨在鉴定和表征乙酰肝素酶剪接变体。LEADS，Compugen的可变剪接建模平台（Compugen，特拉维夫，以色列），用于计算机模拟搜索剪接变体;肿瘤衍生的细胞系（即，CAG骨髓瘤）和肿瘤活组织检查用于验证体内T5表达;信号传导（即，在T5基因沉默或过表达后评估了细胞增殖、集落形成和肿瘤异种移植物发展的相关性（Src磷酸化）。一种新的剪接形式的人乙酰肝素酶，称为T5，被确定。在该剪接变体中，内含子5的144 bp与外显子4连接，这导致截短的、无酶活性的蛋白质。T5过表达导致细胞增殖增加和软琼脂中更大的集落，Src激活介导。此外，T5过表达显著增强肿瘤异种移植物的发展。T5表达上调75%的人肾细胞癌活检检查，这表明这种剪接变异是临床相关的。对照包括过表达野生型乙酰肝素酶或空质粒的细胞和癌病变附近的正常组织。T5是一种新的具有促肿瘤特性的人乙酰肝素酶功能性剪接变体。巴拉什大学，Cohen-Kaplan，V.，Arvatz，G.，Gingis-Velitski，S.，Levy-Adam，F.，Nativ，O.，Shemesh河，Ayalon-Sofer，M.，Ilan，N.，弗洛达夫斯基岛一种新的人乙酰肝素酶剪接变体T5，具有促肿瘤发生的特性。FASEB J.24，1239-1248（2010）。www.fasebj.org
Heparanase is a mammalian endo-beta-D-glucuronidase that can cleave heparan sulfate side chains, an activity strongly implicated in tumor cell dissemination. The current study aimed to identify and characterize heparanase splice variants. LEADS, Compugen's alternative splicing modeling platform (Compugen, Tel Aviv, Israel), was used to search for splice variants in silico; tumor-derived cell lines (i.e., CAG myeloma) and tumor biopsies were utilized to validate T5 expression in vivo; signaling (i.e., Src phosphorylation) was evaluated following T5 gene silencing or overexpression and correlated with cell proliferation, colony formation, and tumor xenograft development. A novel spliced form of human heparanase, termed T5, was identified. In this splice variant, 144 bp of intron 5 are joined with exon 4, which results in a truncated, enzymatically inactive protein. T5 overexpression resulted in increased cell proliferation and larger colonies in soft agar, mediated by Src activation. Furthermore, T5 overexpression markedly enhanced tumor xenograft development. T5 expression is up-regulated in 75% of human renal cell carcinoma biopsies examined, which suggests that this splice variant is clinically relevant. Controls included cells overexpressing wild-type heparanase or an empty plasmid and normal-looking tissue adjacent the carcinoma lesion. T5 is a novel functional splice variant of human heparanase endowed with protumorigenic characteristics.-Barash, U., Cohen-Kaplan, V., Arvatz, G., Gingis-Velitski, S., Levy-Adam, F., Nativ, O., Shemesh, R., Ayalon-Sofer, M., Ilan, N., Vlodavsky, I. A novel human heparanase splice variant, T5, endowed with protumorigenic characteristics. FASEB J. 24, 1239-1248 (2010). www.fasebj.org