Inhibition of UCH-L1 Deubiquitinating Activity with Two Forms of LDN-57444 Has Anti-Invasive Effects in Metastatic Carcinoma Cells
Inhibition of UCH-L1 Deubiquitinating Activity with Two Forms of LDN-57444 Has Anti-Invasive Effects in Metastatic Carcinoma Cells
复制标题
DOI：
10.3390/ijms20153733
复制
发表时间：
2019-08-01
影响因子：
5.6
通讯作者：
Shakelford, Julia
中科院分区：
文献类型：
--
作者：
Kobayashi, Eiji;Hwang, Duhyeong;Shakelford, Julia
泛素C末端水解酶L1(UCH-L1)通常在成人的中枢神经系统和生殖系统中表达，但在许多人类肿瘤中已检测到其从头表达。越来越多的证据表明，UCH-L1去泛素化(DUB)活性在某些癌症中起着主要的促转移作用。在这里，我们测试了UCH-L1 DUB活性的小分子抑制剂LDN-57444在晚期口腔鳞癌细胞系以及表达EB病毒肿瘤病毒主要促转移基因产物LMP1的侵袭性鼻咽癌细胞系中的抗转移作用。为了克服LDN-57444在水中的有限溶解性，我们通过将化合物掺入到聚恶唑啉胶束纳米粒(LDN-POX)中，开发了LDN-57444的纳米制剂。LDN-POX纳米粒在体外的作用与天然化合物相同。我们的结果表明，用LDN或LDN-POX抑制UCH-L1 DUB活性会抑制外体的分泌，并降低外体部分中促转移因子的水平。在生理实验中，两种形式的UCH-L1 DUB抑制剂都抑制转移性鳞癌细胞和表达EBV前转移潜伏膜蛋白1(LMP1)的鼻咽细胞的运动。此外，LDN和LDN-POX治疗导致转移前标志物水平降低，癌细胞粘附性降低，并抑制细胞外小泡(ECV)介导的病毒侵袭因子LMP1的转移。我们认为，可溶性的UCH-L1抑制剂，如LDN-POX，为包括EBV阳性的恶性肿瘤在内的浸润性癌提供了潜在的治疗形式。
Normally ubiquitin C-terminal hydrolase L1 (UCH-L1) is expressed in the central nervous and reproductive systems of adults, but its de novo expression has been detected in many human cancers. There is a growing body of evidence that UCH-L1 de-ubiquitinating (DUB) activity plays a major pro-metastatic role in certain carcinomas. Here we tested anti-metastatic effects of the small-molecule inhibitor of UCH-L1 DUB activity, LDN-57444, in cell lines from advanced oral squamous cell carcinoma (OSCC) as well as invasive nasopharyngeal (NP) cell lines expressing the major pro-metastatic gene product of Epstein-Barr virus (EBV) tumor virus, LMP1. To overcome the limited aqueous solubility of LDN-57444 we developed a nanoparticle formulation of LDN-57444 by incorporation of the compound in polyoxazoline micellear nanoparticles (LDN-POx). LDN-POx nanoparticles were equal in effects as the native compound in vitro. Our results demonstrate that inhibition of UCH-L1 DUB activity with LDN or LDN-POx inhibits secretion of exosomes and reduces levels of the pro-metastatic factor in exosomal fractions. Both forms of UCH-L1 DUB inhibitor suppress motility of metastatic squamous carcinoma cells as well as nasopharyngeal cells expressing EBV pro-metastatic Latent membrane protein 1 (LMP1) in physiological assays. Moreover, treatment with LDN and LDN-POx resulted in reduced levels of pro-metastatic markers, a decrease of carcinoma cell adhesion, as well as inhibition of extra-cellular vesicle (ECV)-mediated transfer of viral invasive factor LMP1. We suggest that soluble inhibitors of UCH-L1 such as LDN-POx offer potential forms of treatment for invasive carcinomas including EBV-positive malignancies.