Importin-11 overexpression promotes the migration, invasion, and progression of bladder cancer associated with the deregulation of CDKN1A and THBS1
Importin-11 overexpression promotes the migration, invasion, and progression of bladder cancer associated with the deregulation of CDKN1A and THBS1
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Importin-11 过表达促进膀胱癌的迁移、侵袭和进展，与 CDKN1A 和 THBS1 的失调相关
DOI：
10.1016/j.urolonc.2018.03.001
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发表时间：
2018-06-01
影响因子：
2.7
通讯作者：
Xu, Chuanliang
中科院分区：
文献类型：
--
作者：
Zhao, Junjie;Shi, Lei;Xu, Chuanliang
目的：我们最近确定了一个新的癌基因，IPO 11从5 q12，参与膀胱癌（BCa）的进展。然而，IPO 11的生物学功能及其促进BCa进展的分子机制仍不清楚。本研究的目的是调查的作用，IPO 11在BCa的侵略性和阐明其影响的分子机制在BCa.Materials和方法：IPO 11的mRNA表达水平在BIU-87，RT 4，UMUC 3，EJ，5637，T24，J82，HT-1376细胞系采用实时定量聚合酶链反应。采用免疫组化方法检测134例福尔马林固定石蜡包埋（FFPE）的膀胱癌组织和10例配对的非肿瘤性膀胱组织中importin-11的表达。采用荧光原位杂交技术检测了25例FFPE BCa标本中IPO 11的拷贝数。使用RNA干扰在EJ和5637细胞系中研究了IPO 11对迁移、侵袭和细胞增殖的影响。在EJ细胞和IPO 11沉默的EJ细胞中使用全转录组测序和生物信息学方法研究了潜在的分子机制，并使用定量实时聚合酶链反应进行了验证。6种侵袭性BCa细胞系均高表达内源性IPO 11 mRNA（EJ，HT-1376，UMUC3，5637，J82，免疫组化染色显示，87在134例FFPE BCa组织中，importin-11的表达率为64.9%。此外，importin-11过表达与肿瘤分期和肿瘤分级、淋巴浸润和淋巴结转移呈正相关。此外，在100%（14/14）的具有IPO 11扩增的BCa组织中检测到importin-11过表达，并且在另外2个具有importin-11过表达的BCa组织中未观察到IPO 11扩增。小干扰RNA介导的IPO 11敲低足以抑制EJ和5637细胞的运动性和侵袭性。IPO 11敲低也抑制了EJ细胞中的细胞增殖，而这在5637细胞或体内实验中没有观察到。使用全转录组测序，我们发现22个基因（包括IPO 11）在IPO 11沉默的EJ细胞与野生型EJ细胞相比差异表达，其中4个上调，18个下调。显著差异表达基因的KEGG途径富集分析显示，在10个遗传改变的途径中，癌症途径（途径Id：hsa 05205）中的蛋白聚糖富集最显著，并且涉及6个显著改变的基因（CDKN 1A、HBEGF、PTK 2、THBS 1、CCNG 2和EGR 1）。接下来的3个最显著富集的途径依次是p53、ErbB和BCa途径。CDKN 1A和THBS 1是两个最常见的基因，分别参与9条和6条通路。结论：Importin-11过表达可促进BCa细胞的侵袭能力，其机制可能与CDKN 1A和THBS 1的表达失调有关，主要通过激活癌细胞通路和经典BCa通路中的蛋白多糖。Importin-11可能是一个有用的靶点，通过它可以阻止非侵袭性BCa向侵袭性BCa的进展。(C)2018爱思唯尔公司All rights reserved.
Objectives: We recently determined that a novel oncogene, IPO11 from 5q12, participates in bladder cancer (BCa) progression. However, the biological function of IPO11 and the molecular mechanisms through which it contributes to BCa progression remain unclear. The aim of this study was to investigate the role of IPO11 in BCa aggressiveness and elucidate the molecular mechanisms underlying its effects in BCa.Materials and methods: The mRNA expression levels of IPO11 in BIU-87, RT4, UMUC3, EJ, 5637, T24, J82, and HT-1376 cell lines were determined using quantitative real-time polymerase chain reaction. Expression of importin-11 was detected in 134 formalin-fixed and paraffin-embedded (FFPE) BCa tissues and 10 paired nonneoplastic bladder tissue specimens by immunohistochemistry. The copy number of IPO11 was examined in 25 FFPE BCa specimens using fluorescent in situ hybridization. The effects of IPO11 on migration, invasion, and cell proliferation were investigated in EJ and 5637 cell lines using RNA interference. Potential molecular mechanisms were investigated using whole transcriptome sequencing and bioinformatic approaches in EJ cells and IPO11-silenced EJ cells and verified using quantitative real-time polymerase chain reaction.Results: Endogenous IPO11 mRNA was highly expressed in 6 invasive BCa cell lines (EJ, HT-1376, UMUC3, 5637, J82, and T24) but had a low expression in the noninvasive BCa cell line BIU-87 and the papillary BCa cell line RT4 Immunohistochemical staining revealed that 87 (64.9%) of 134 FFPE BCa tissues displayed importin-11 overexpression. Moreover, importin-11 overexpression was positively associated with increased tumor stages and tumor grades, lymphatic invasion, and lymph node metastasis. Furthermore, importin-11 overexpression was detected in 100% (14/14) of BCa tissues with IPO11 amplification, and IPO11 amplification was not observed in 2 additional BCa tissues with importin-11 overexpression. Small interfering RNA-mediated knockdown of IPO11 is sufficient to inhibit the motility and invasiveness of EJ and 5637 cells. IPO11 knockdown also inhibited cell proliferation in EJ cells, whereas this was not observed in 5637 cells or the in vivo experiments. Using whole transcriptome sequencing, we found that 22 genes (including IPO11) were differentially expressed in IPO11-silenced EJ cells compared with wild-type EJ cells, 4 of which were upregulated, and 18 of which were downregulated. KEGG pathway enrichment analysis of the significantly differentially expressed genes showed that the proteoglycans in cancer pathway (pathway Id: hsa05205) was most significantly enriched among 10 genetically altered pathways and referred to 6 significantly altered genes (CDKN1A, HBEGF, PTK2, THBS1, CCNG2, and EGR1). The next 3 most significantly enriched pathways in order were the p53, ErbB, and BCa pathways. CDKN1A and THBS1 were the most 2 frequently covered genes and were involved in 9 and 6 pathways, respectively. They were also 2 key proteins in the BCa pathway (pathway Id: hsa05219) that were downregulated in IPO11-knockdown EJ cells compared with wild-type EJ cells.Conclusions: Importin-11 overexpression can promote BCa cell invasiveness, probably associated with the deregulation of CDKN1A and THBS1 primarily through the activation of the proteoglycans in cancer pathway and the classical BCa pathway. Importin-11 may be a useful target through which the progression of noninvasive BCa to invasive BCa can be blocked. (C) 2018 Elsevier Inc. All rights reserved.