Exosomal Circ-ZNF652 Promotes Cell Proliferation, Migration, Invasion and Glycolysis in Hepatocellular Carcinoma via miR-29a-3p/GUCD1 Axis
Exosomal Circ-ZNF652 Promotes Cell Proliferation, Migration, Invasion and Glycolysis in Hepatocellular Carcinoma via miR-29a-3p/GUCD1 Axis
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DOI：
10.2147/cmar.s259424
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发表时间：
2020-01-01
影响因子：
3.3
通讯作者：
Huang, Guomin
中科院分区：
文献类型：
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作者：
Li, Yuhui;Zang, Hongliang;Huang, Guomin
背景：环状rna （circRNAs）在肝细胞癌（HCC）的进展中起着至关重要的作用。然而，外泌体环状rna在HCC中的作用在很大程度上仍然未知。我们旨在探讨外泌体circ-ZNF652在HCC中的功能。方法：采用透射电子显微镜（TEM）和纳米颗粒跟踪分析（NTA）检测外泌体的形态和大小。采用实时荧光定量聚合酶链式反应（qRT-PCR）检测circZNF652、ZNF652 mRNA、microRNA-29a-3p （miR-29a-3p）和含1冠酰环化酶结构域（GUCD1） mRNA的表达。Western blot检测CD63、CD81、己糖激酶2 （HK2）和GUCD1蛋白水平。采用RNase R酶切法检测circ-ZNF652的稳定性。采用3-（4,5 -二甲基-2-噻唑基）- 2,5 -二苯基-2- h -溴化四唑（MTT）法检测细胞增殖情况。transwell法检测细胞迁移和侵袭。通过特异性试剂盒检测糖酵解水平。通过双荧光素酶报告基因法和RNA免疫沉淀（RIP）法分析miR-29a-3p与circ-ZNF652或GUCD1的相关性。建立小鼠异种移植瘤模型，探讨circ-ZNF652在体内的作用。结果：外泌体circ-ZNF652在HCC患者血清和HCC细胞中表达上调。外泌体circ-ZNF652可转移至HCC细胞，沉默circ-ZNF652可抑制HCC细胞的增殖、迁移、侵袭和糖酵解。Circ-ZNF652是miR-29a-3p的海绵，miR-29a-3p抑制剂减弱了Circ-ZNF652沉默对HCC细胞进展的抑制作用。GUCD1是miR-29a-3p的靶基因，GUCD1过表达恢复了miR-29a-3p对HCC细胞发育的影响。此外，circZNF652基因敲低在体内抑制肿瘤生长。结论：外泌体circ-ZNF652通过miR-29a-3p/GUCD1轴参与HCC细胞增殖、迁移、侵袭和糖酵解。
Background: Circular RNAs (circRNAs) play a crucial role in hepatocellular carcinoma (HCC) progression. However, the role of exosomal circRNAs in HCC is still largely unknown. We aimed to explore the function of exosomal circ-ZNF652 in HCC.Methods: The morphology and size of exosomes were examined by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA). The expression of circZNF652, ZNF652 mRNA, microRNA-29a-3p (miR-29a-3p) and guanylyl cyclase domain containing 1 (GUCD1) mRNA was determined by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of CD63, CD81, hexokinase 2 (HK2) and GUCD1 were examined via Western blot assay. The stability of circ-ZNF652 was examined by RNase R digestion assay. Cell proliferation was analyzed by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Cell migration and invasion were assessed by transwell assay. The glycolysis level was detected via specific kits. The association between miR-29a-3p and circ-ZNF652 or GUCD1 was analyzed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. A murine xenograft model was constructed to explore the effect of circ-ZNF652 in vivo.Results: Exosomal circ-ZNF652 was upregulated in HCC patients' serums and HCC cells. Exosomal circ-ZNF652 could transfer to HCC cells, and circ-ZNF652 silencing suppressed HCC cell proliferation, migration, invasion and glycolysis. Circ-ZNF652 was a sponge of miR-29a-3p, and the inhibitory effect of circ-ZNF652 silencing on HCC cell progression was weakened by miR-29a-3p inhibitor. GUCD1 was a target gene of miR-29a-3p, and GUCD1 overexpression restored the effect of miR-29a-3p on HCC cell development. Moreover, circZNF652 knockdown repressed tumor growth in vivo.Conclusion: Exosomal circ-ZNF652 contributes to HCC cell proliferation, migration, invasion and glycolysis by miR-29a-3p/GUCD1 axis.