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Abstract
BackgroundHepatocellular carcinoma (HCC) is the most common malignant heterogeneous disease in primary liver tumors. Circular RNA hsa_circ_0000517 (hsa_circ_0000517) is connected with HCC prognosis. Nevertheless, there are few studies on the role and mechanism of hsa_circ_0000517 in HCC.MethodsExpression of hsa_circ_0000517, miR-326, and SMAD family member 6 (SMAD6) was detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell viability, colony formation, cell cycle, migration, and invasion were determined though Cell Counting Kit-8 (CCK-8), colony formation, flow cytometry, wound healing, or transwell assays. Protein levels of Cyclin D1, matrix metalloproteinase-2 (MMP2), matrix metalloproteinase-9 (MMP9), SMAD6, and proliferating cell nuclear antigen (PCNA) were examined with western blot analysis. The relationship between hsa_circ_0000517 or SMAD6 and miR-326 was determined via dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. The role of hsa_circ_0000517 in vivo was confirmed via xenograft assay.ResultsHsa_circ_0000517 and SMAD6 were up-regulated while miR-326 was down-regulated in HCC tissues and cells. Hsa_circ_0000517 down-regulation repressed cell proliferation, colony formation, migration, and invasion, and induced cell cycle arrest in HCC cells in vitro, and constrained tumor growth in vivo. Notably, hsa_circ_0000517 regulated SMAD6 expression via acting as a competing endogenous RNA (ceRNA) for miR-326. And the repressive influence on malignant behaviors of HCC cells mediated by hsa_circ_0000517 inhibition was reversed by miR-326 inhibitors. Moreover, SMAD6 elevation overturned the inhibitory impacts of miR-326 mimics on malignant behaviors of HCC cells.ConclusionsHsa_circ_0000517 depletion repressed HCC advancement via regulating the miR-326/SMAD6 axis.
Highlights
Hepatocellular carcinoma (HCC) is the most common malignant heterogeneous disease in primary liver tumors
Patients and specimens All experimental protocols in this research were ratified by the Ethics Committee of the First Affiliated Hospital of Zhengzhou University. 50 paired HCC tissues and adjoining normal tissues were obtained from HCC patients who underwent surgery at the First Affiliated Hospital of Zhengzhou University
Silence of hsa_circ_0000517 repressed cell proliferation, colony formation, migration, and invasion, and induced cell cycle arrest in HCC cells In view of the enhancement of hsa_circ_0000517 in HCC tissues and cells, we investigated the role of hsa_circ_0000517 in HCC via loss-of-function experiments
Summary
Hepatocellular carcinoma (HCC) is the most common malignant heterogeneous disease in primary liver tumors. Hepatocellular carcinoma (HCC), the most common malignant heterogeneous disease in primary liver tumors, ranks fourth among cancer-related causes of death [1]. The treatment of HCC mainly includes surgery, He et al Cancer Cell Int (2020) 20:360 radio-therapy, or chemo-therapy [2, 3]. Despite advances in the diagnosis of HCC, only 30–40% of patients can be treated with surgery [4]. Most HCC patients have metastases and relapses within 5 years after undergoing surgical treatment [7, 8]. It is vital to survey the mechanisms related to HCC progression for developing new diagnostic biomarkers and treatment strategies
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