JunB promotes cell invasion and angiogenesis in VHL-defective renal cell carcinoma
JunB promotes cell invasion and angiogenesis in VHL-defective renal cell carcinoma
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DOI：
10.1038/onc.2011.475
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发表时间：
2012-06-01
期刊：
影响因子：
8
通讯作者：
Nakamura, E.
中科院分区：
文献类型：
--
作者：
Kanno, T.;Kamba, T.;Nakamura, E.
Von Hippel-Lindau(VHL)抑癌基因失活导致遗传性和散发性肾透明细胞癌(CcRCC)。尽管VHL蛋白(PVHL)最具特色的功能是调节缺氧诱导因子-α(HIF-α)，但pVHL也通过调节JunB，通过非HIF途径控制嗜铬细胞瘤的发展。然而，很大程度上还不清楚这些通路如何促进ccRCC的发展和进展。在本研究中，我们通过免疫染色证实了JunB在VHL缺陷的ccRCC标本中表达上调。短发夹状RNA(ShRNA)介导的JunB基因敲除786-O和A498VHL缺失的ccRCC细胞可抑制其侵袭力。此外，在异种移植瘤检测中，JunB基因敲除显著抑制了肿瘤生长和微血管密度。相反，在VHL修复的786-O亚克隆中强制表达野生型而不是二聚化缺陷的JunB可以促进体外侵袭和体内肿瘤生长和血管形成。定量PCR芯片分析显示，JunB在786-O细胞中调控与肿瘤侵袭和血管生成相关的多个基因，如基质金属蛋白酶-2(MMP2)、MMP9和趋化因子(C-C基序)配体2(CCL2)。在这些细胞中，JunB基因敲除降低了明胶酶谱中两种MMPs的蛋白分解活性和培养上清液中CCL2的量。此外，shRNA介导的抑制基质金属蛋白酶-2或用中和抗体抑制CCL2活性抑制了异种移植瘤的生长和血管生成。综上所述，这些结果表明，JunB促进了VHL缺陷的CCRCC的肿瘤侵袭力和血管生成。Oncogene(2012年)31,3098-3110；doi：10.1038/onc.2011.475；2011年10月24日在线发布
Inactivation of the von Hippel-Lindau (VHL) tumor-suppressor gene causes both hereditary and sporadic clear-cell renal-cell carcinoma (ccRCC). Although the best-characterized function of the VHL protein (pVHL) is regulation of hypoxia-inducible factor-alpha (HIF alpha), pVHL also controls the development of pheochromocytoma through HIF-independent pathways by regulating JunB. However, it is largely unknown how these pathways contribute to the development and progression of ccRCC. In the present study, we confirmed that JunB was upregulated in VHL-defective ccRCC specimens by immunostaining. Short-hairpin RNA (shRNA)-mediated knockdown of JunB in 786-O and A498 VHL null ccRCC cells suppressed their invasiveness. In addition, JunB knockdown significantly repressed tumor growth and microvessel density in xenograft tumor assays. Conversely, forced expression of wild-type, but not dimerization-defective, JunB in a VHL-restored 786-O subclone promoted invasion in vitro and tumor growth and vessel formation in vivo. Quantitative PCR array analysis revealed that JunB regulated multiple genes relating to tumor invasion and angiogenesis such as matrix metalloproteinase-2 (MMP-2), MMP-9 and chemokine (C-C motif) ligand-2 (CCL2) in 786-O cells. JunB knockdown in these cells reduced the proteolytic activity of both MMPs in gelatin zymography and the amount of CCL2 in the culture supernatant. Moreover, shRNA-mediated knockdown of MMP-2 or inhibition of CCL2 activity with a neutralizing antibody repressed xenograft tumor growth and angiogenesis. Collectively, these results suggest that JunB promotes tumor invasiveness and enhances angiogenesis in VHL-defective ccRCCs. Oncogene (2012) 31, 3098-3110; doi: 10.1038/onc.2011.475; published online 24 October 2011