MIIP inhibits clear cell renal cell carcinoma proliferation and angiogenesis via negative modulation of the HIF-2α-CYR61 axis.
MIIP inhibits clear cell renal cell carcinoma proliferation and angiogenesis via negative modulation of the HIF-2α-CYR61 axis.
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MIIP 通过 HIF-2α-CYR61 轴的负调节抑制透明细胞肾细胞癌增殖和血管生成
DOI：
10.20892/j.issn.2095-3941.2020.0296
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发表时间：
2021-12-22
影响因子：
5.5
通讯作者：
Li, Xia
中科院分区：
文献类型：
--
作者：
Yan, Fengqi;Wang, Qinhao;Xia, Mingyuan;Ru, Yi;Hu, Wei;Yan, Guang;Xiong, Xin;Zhang, Mei;Wang, Jiancai;Li, Qi;Zhang, Bo;Wang, He;Lin, Wei;Wu, Guojun;Li, Xia
目的：在多种肿瘤中，迁移和侵袭抑制蛋白（MIIP）呈低水平表达，并参与肿瘤的发病机制。在此，我们试图探索MIIP在肾透明细胞癌（ccRCC）中的功能。研究方法：CCK-8、集落形成、细胞周期和内皮细胞管形成测定用于评估MIIP在ccRCC增殖和血管生成中的作用。为了探索潜在的机制，我们在ccRCC细胞系中进行了RNA测序、GSEA、qRT-PCR、Western印迹、ELISA、细胞转染、免疫共沉淀和泛素化测定。此外，检查裸鼠中的异种移植肿瘤生长以及异种移植组织中的Ki-67和CD 31染色。最后，通过Western blot和免疫组化方法进一步分析MIIP在肾癌组织中的表达与临床病理的关系，以及HIF-2α和CYR 61在肾癌组织中的表达情况。结果如下：体内外功能实验均表明，强制表达MIIP可抑制ccRCC细胞增殖和血管生成，而沉默正常HK-2细胞和ccRCC细胞中的MIIP则具有相反的作用（P < 0.05）。在机制上，CYR 61被鉴定为通过MIIP过表达显著下调的基因，并且是MIIP在ccRCC中的抑制作用所需的。发现MIIP促进HSP 90乙酰化，从而损害其对HIF-2α的伴侣功能。因此，RACK 1结合HIF-2α并导致其泛素化和蛋白酶体降解，从而降低其靶CYR 61的转录。最后，临床标本分析表明，MIIP在肾癌组织中表达显著下调，其表达与肾癌组织学分级、转移、预后以及HIF-2α和CYR 61的表达呈负相关（P < 0.05）。结论：MIIP可能通过负调控HIF-2α-CYR 61轴在ccRCC中发挥抑癌作用。
Objective: In various cancers, migration and invasion inhibitory protein (MIIP) is expressed at low level and is involved in cancer pathogenesis. Herein, we sought to explore the function of MIIP in clear cell renal cell carcinoma (ccRCC). Methods: CCK-8, colony formation, cell cycle, and endothelial cell tube formation assays were performed to evaluate the roles of MIIP in ccRCC proliferation and angiogenesis. To explore the underlying mechanism, we conducted RNA-sequencing, GSEA, qRT-PCR, Western blot, ELISA, cell transfection, coimmunoprecipitation, and ubiquitination assays in ccRCC cell lines. Furthermore, xenograft tumor growth in nude mice, and Ki-67 and CD31 staining in xenograft tissues were examined. Finally, the association of MIIP expression with clinical pathology and the expression status of HIF-2α and cysteine-rich 61 (CYR61) were further analyzed in human RCC tissues through Western blot and immunohistochemistry. Results: Both in vitro and in vivo functional experiments indicated that forced expression of MIIP inhibited ccRCC proliferation and angiogenesis, whereas silencing MIIP either in normal HK-2 cells or in ccRCC cells had the opposite effect (P < 0.05). Mechanistically, CYR61 was identified as a gene significantly downregulated by MIIP overexpression, and was required for the suppressive role of MIIP in ccRCC. MIIP was found to promote HSP90 acetylation and thus impair its chaperone function toward HIF-2α. Consequently, RACK1 binds HIF-2α and causes its ubiquitination and proteasomal degradation, thus decreasing the transcription of its target, CYR61. Finally, analyses of clinical samples demonstrated that MIIP is significantly downregulated in cancer vs. normal tissues in RCC cases, and its expression is negatively associated with histological grade, metastasis, the prognosis of patients with RCC, and the expression of HIF-2α and CYR61 (P < 0.05). Conclusions: MIIP is a novel tumor suppressor in ccRCC via negative regulation of HIF-2α-CYR61 axis.
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