Knockdown of circDENND4C inhibits glycolysis, migration and invasion by up-regulating miR-200b/c in breast cancer under hypoxia
Knockdown of circDENND4C inhibits glycolysis, migration and invasion by up-regulating miR-200b/c in breast cancer under hypoxia
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DOI：
10.1186/s13046-019-1398-2
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发表时间：
2019-09-05
影响因子：
11.3
通讯作者：
Zhang, Yanwu
中科院分区：
文献类型：
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作者：
Ren, Shasha;Liu, Jiuzhou;Zhang, Yanwu
研究背景低氧是乳腺癌的一个重要特征，影响肿瘤的发生、转移和代谢。以往的研究表明，环状RNA(CircRNAs)可能参与肿瘤进展和缺氧调节。本研究旨在探讨在正常细胞和含有4C的肿瘤结构域(CircDENND4C)中差异表达的CircRNA在低氧条件下乳腺癌进展中的作用。方法分析43例乳腺癌患者的临床资料。将乳腺癌细胞株MDA-MB-453和SK-BR-3在低氧(1%O-2)条件下进行体外培养。采用实时荧光定量聚合酶链式反应技术检测细胞中CircDENND4C、microRNA-200b(miR-200b)和miR-200C的表达水平。糖酵解以葡萄糖消耗、乳酸产量和己糖激酶II(HK2)蛋白水平为指标。通过跨孔分析和基质金属肽酶9(MMP9)和MMP2的蛋白水平评价肿瘤的迁移和侵袭。通过生物信息学分析、荧光素酶活性测定和RNA免疫沉淀法研究了CircDENND4C与miR-200b或miR-200C之间的相互作用。建立小鼠异种移植模型，研究CircDENND4C的体内抗癌作用。结果DNND4C在乳腺癌组织中的高表达随低氧反应而上调。DNND4C基因敲除减少了低氧条件下乳腺癌细胞的糖酵解、迁移和侵袭。CircDENND4C是miR-200b和miR-200C的海绵。MiR-200b或miR-200C的缺失逆转了CircDENND4C基因敲除对乳腺癌进展的抑制作用。此外，沉默周围的DENND4C通过增加miR-200b和miR-200C而抑制移植瘤的生长。结论DENND4C沉默通过上调miR-200b和miR-200C抑制低氧条件下乳腺癌细胞的糖酵解、迁移和侵袭。
Background Hypoxia is a key feature of breast cancer, which affects cancer development, metastasis and metabolism. Previous studies suggested that circular RNAs (circRNAs) could participate in cancer progression and hypoxia regulation. This study aimed to investigate the role of circRNA differentially expressed in normal cells and neoplasia domain containing 4C (circDENND4C) in breast cancer progression under hypoxia. Methods Forty-three patients with breast cancer were involved in this study. Breast cancer cell lines MDA-MB-453 and SK-BR-3 were cultured under hypoxia (1% O-2) for experiments in vitro. The expression levels of circDENND4C, microRNA-200b (miR-200b) and miR-200c were measured by quantitative real-time polymerase chain reaction. Glycolysis was investigated by glucose consumption, lactate production and hexokinase II (HK2) protein level. Migration and invasion were evaluated via trans-well assay and protein levels of matrix metallopeptidase 9 (MMP9) and MMP2. The interaction between circDENND4C and miR-200b or miR-200c was explored by bioinformatics analysis, luciferase assay and RNA immunoprecipitation. Murine xenograft model was established to investigate the anti-cancer role of circDENND4C in vivo. Results circDENND4C highly expressed in breast cancer was up-regulated in response to hypoxia. Knockdown of circDENND4C decreased glycolysis, migration and invasion in breast cancer cells under hypoxia. circDENND4C was validated as a sponge of miR-200b and miR-200c. Deficiency of miR-200b or miR-200c reversed the suppressive effect of circDENND4C knockdown on breast cancer progression. Moreover, silence of circDENND4C reduced xenograft tumor growth by increasing miR-200b and miR-200c. Conclusion circDENND4C silence suppresses glycolysis, migration and invasion in breast cancer cells under hypoxia by increasing miR-200b and miR-200c.