miR-497-5p inhibits cell proliferation and invasion by targeting KCa3.1 in angiosarcoma.
miR-497-5p inhibits cell proliferation and invasion by targeting KCa3.1 in angiosarcoma.
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miR-497-5p通过靶向KCa3.1抑制血管肉瘤细胞增殖和侵袭
DOI：
10.18632/oncotarget.11252
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发表时间：
2016-09-06
期刊：
影响因子：
--
通讯作者：
Duan Y
中科院分区：
文献类型：
--
作者：
Chen Y;Kuang D;Zhao X;Chen D;Wang X;Yang Q;Wan J;Zhu Y;Wang Y;Zhang S;Wang Y;Tang Q;Masuzawa M;Wang G;Duan Y
血管肉瘤是一种少见的恶性间充质肿瘤，预后差。我们的目的是鉴定与肿瘤相关的miRNAs及其靶基因，并探讨miRNA及其靶基因在血管肉瘤中的生物学功能。通过miRNA芯片和逆转录聚合酶链式反应，我们在人血管肉瘤中发现了1个上调的miRNA(miR-222-3p)和3个下调的miRNAs(miR-497-5p、miR-378-3p和miR-483-5p)。中电导钙激活钾通道KCa3.1是miR-497-5p的可能靶基因之一，免疫组织化学检测到KCa3.1在血管肉瘤活检标本中显著上调。在ISO-HAS血管肉瘤细胞系中，miR-497-5p和KCa3.1在mRNA和蛋白水平上也呈负相关。MiR-497-5p与KCa3.1mRNA3‘非翻译区的互补结合降低了荧光素酶活性，证明了miR-497-5p直接靶向KCa3.1。对于miR-497-5p/KCa3.1对的功能作用，我们发现，应用特异性KCa3.1通道阻断剂TRAM-34，或将KCa3.1 siRNA或miR-497-5p模拟物导入ISO-HAS细胞，可通过下调细胞周期相关蛋白，包括细胞周期蛋白D1、Surviving和p53以及下调基质金属肽酶9来抑制细胞增殖、细胞周期进展和侵袭。在体内血管肉瘤移植瘤模型中，TRAM-34或miR-497-5p模拟物均可抑制肿瘤生长。总之，肿瘤抑制基因miR-497-5p下调KCa3.1的表达，有助于抑制血管肉瘤的恶性发展。MiR-497-5p或KCa3.1可能成为治疗血管肉瘤的新靶点。
Angiosarcoma is a rare malignant mesenchymal tumor with poor prognosis. We aimed to identify malignancy-associated miRNAs and their target genes, and explore biological functions of miRNA and its target in angiosarcoma. By miRNA microarrays and reverse transcription polymerase chain reaction, we identified 1 up-regulated miRNA (miR-222-3p) and 3 down-regulated miRNAs (miR-497-5p, miR-378-3p and miR-483-5p) in human angiosarcomas compared with human capillary hemangiomas. The intermediate-conductance calcium activated potassium channel KCa3.1 was one of the putative target genes of miR-497-5p, and marked up-regulation of KCa3.1 was detected in angiosarcoma biopsy specimens by immunohistochemistry. The inverse correlation of miR-497-5p and KCa3.1 also was observed in the ISO-HAS angiosarcoma cell line at the mRNA and protein levels. The direct targeting of KCa3.1 by miR-497-5p was evidenced by reduced luciferase activity due to complementary binding of miR-497-5p to KCa3.1 mRNA 3′ untranslated region. For the functional role of miR-497-5p/KCa3.1 pair, we showed that application of TRAM-34, a specific KCa3.1 channel blocker, or transfection of ISO-HAS cells with KCa3.1 siRNA or miR-497-5p mimics inhibited cell proliferation, cell cycle progression, and invasion by down-regulating cell-cycle related proteins including cyclin D1, surviving and P53 and down-regulating matrix metallopeptidase 9. In an in vivo angiosarcoma xenograft model, TRAM-34 or miR-497-5p mimics both inhibited tumor growth. In conclusion, the tumor suppressor miR-497-5p down-regulates KCa3.1 expression and contributes to the inhibition of angiosarcoma malignancy development. The miR-497-5p or KCa3.1 might be potential new targets for angiosarcoma treatment.