USP13 functions as a tumor suppressor by blocking the NF-kB-mediated PTEN downregulation in human bladder cancer
USP13 functions as a tumor suppressor by blocking the NF-kB-mediated PTEN downregulation in human bladder cancer
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USP13 通过阻断人类膀胱癌中 NF-kB 介导的 PTEN 下调来发挥肿瘤抑制因子的作用
DOI：
10.1186/s13046-019-1262-4
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发表时间：
2019-06-14
影响因子：
11.3
通讯作者：
Jiang, Yuanjun
中科院分区：
文献类型：
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作者：
Man, Xiaojun;Piao, Chiyuan;Jiang, Yuanjun
背景USP 13与人类肿瘤的发生发展密切相关，但其在膀胱癌中的作用及调控机制尚不清楚。方法采用逆转录-聚合酶链反应（RT-PCR）技术检测miR-130 b-3 p、miR-301 b-3 p和USP 13在膀胱癌组织中的表达。通过Western印迹、q-RT-PCR、生物信息学分析和双荧光素酶报告基因测定来鉴定miR-130 b-3 p/301 b-3 p对USP 13的调节功能。采用免疫共沉淀法检测USP 13与PTEN蛋白的相互作用。采用流式细胞仪、集落形成实验和transwell实验检测BC细胞的增殖、迁移和侵袭能力。建立BC细胞的小鼠异种移植模型，以验证USP 13在体内的功能。免疫组化进行鉴定的蛋白表达的USP 13，NF-κ B p65或PTEN在临床/异种移植肿瘤tissues.ResultsOur目前的研究表明，USP 13的功能作为一个肿瘤抑制因子，通过相互作用与PTEN蛋白，并增加其在膀胱癌中的表达。我们发现USP 13的缺失导致PTEN的下调，并促进膀胱癌细胞的增殖、侵袭和迁移能力。此外，我们发现USP 13是miR-130 b-3 p和miR-301 b-3 p以及miR-130 b/301 b簇的共同靶点，其可以被NF-κ B转录上调。我们的数据表明，NF-κ B活化降低USP 13和PTEN的表达水平，并促进BC细胞的成瘤表型。此外，重新引入USP 13可部分挽救PTEN的表达以及NF-κ B引起的肿瘤发生趋势。结论NF-κ B诱导的miR-130 b/301 b过表达降低了USP 13的表达，进而导致PTEN蛋白表达下调，促进了膀胱癌的发生。此外，NF-κ B介导的PTEN下调很可能促进NF-κ B的完全激活。
BackgroundUSP13 has been reported to be involved in the tumorigenesis of human cancers, however, its functional role and regulatory mechanisms in bladder cancer (BC) remain unclear.Methodsq-RT-PCR was performed to examine the expression of miR-130b-3p, miR-301b-3p and USP13 in BC tissue samples. Western blot, q-RT-PCR, bioinformatic analysis and dual-luciferase reporter assay were conducted to identify the regulatory function of miR-130b-3p/301b-3p for USP13. Co-immunoprecipitation assay was performed to assess the interaction between USP13 and PTEN protein. Cell-counting-kit 8, colony formation assay and transwell assay were performed to value the proliferative, migrative and invasive capacities of BC cells in vitro. Mouse xenograft model of BC cells was established to verify the function of USP13 in vivo. Immunohistochemistry was performed to identify the protein expression of USP13, NF-kB p65 or PTEN in clinical/xenograft tumor tissues.ResultsOur present study reveals that USP13 functions as a tumor suppressor by interacting with PTEN protein and increasing its expression in bladder cancer. We found that loss of USP13 led to the downregulation of PTEN and promoted proliferative, invasive and migrative capacities of bladder cancer cells. Furthermore, we discovered that USP13 was a common target of miR-130b-3p and miR-301b-3p, and the miR-130b/301b cluster, which could be transcriptionally upregulated by NF-kB. Our data demonstrated that NF-kB activation decreased expression level of USP13 and PTEN, and promoted the tumorigenesis phenotypes of BC cells. In addition, reintroduction of USP13 partially rescued PTEN expression as well as the oncogenesis trend caused by NF-kB.ConclusionWe reported a potential regulatory loop that the NF-kB-induced miR-130b/301b overexpression decreased USP13 expression and subsequently resulted in the downregulation of PTEN protein and promoted tumorigenesis of bladder cancer. Moreover, NF-kB-mediated PTEN downregulation is very likely to facilitate the full activation of NF-kB.