Abstract
New evidence suggests that abnormal expression of circular RNA (circRNA) is associated with the development of human cancers. This study aims to reveal circMYOF roles in the malignant phenotype of laryngeal squamous cell carcinoma (LSCC). The expression of circMYOF, microRNA (miR)-145-5p, and orthodenticle homeobox 1 (OTX1) was detected by quantitative real-time PCR. Cell proliferation, migration, invasion, and apoptosis were determined using colony formation assay and EdU assay, wound healing assay, transwell assay, and flow cytometry, respectively. Protein expression was examined by western blot analysis. Cell glycolysis was assessed by detecting glucose consumption and lactate production. Mice xenograft models were constructed to evaluate the regulation of circMYOF on LSCC tumorigenesis. The regulatory relationships among circMYOF, miR-145-5p, and OTX1 were identified using dual-luciferase reporter assay and RIP assay. Serum exosomes were isolated to confirm the existence of circMYOF in LSCC patients. CircMYOF was upregulated in LSCC tissues and cells, and its knockdown suppressed LSCC cell growth, metastasis, and glycolysis, as well as inhibited LSCC tumor growth. MiR-145-5p had decreased expression in LSCC, and it could be sponged by circMYOF. The inhibition effect of circMYOF lentivirus short hairpin RNA (sh-circMYOF) on LSCC progression was restored by the inhibitor of miR-145-5p (in-miR-145-5p). Also, OTX1 was targeted by miR-145-5p and was positively regulated by circMYOF. MiR-145-5p could repress LSCC progression, and OTX1 overexpression also eliminated this effect. In addition, we found that circMYOF was significantly overexpressed in the serum exosomes of LSCC patients. Our data revealed that circMYOF contributed to LSCC progression by promoting cell growth, metastasis, and glycolysis through miR-145-5p/OTX1 axis.
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References
Al-Hakami HA et al (2021) The incidence of thyroid gland invasion in advanced laryngeal squamous cell carcinoma. Braz J Otorhinolaryngol 87(5):533–537
Fong PY et al (2019) Association of clinical factors with survival outcomes in laryngeal squamous cell carcinoma (LSCC). PLoS ONE 14(11):e0224665
Chen H et al (2020a) Preoperative platelet-lymphocyte ratio predicts recurrence of laryngeal squamous cell carcinoma. Future Oncol 16(6):209–217
Cui J et al (2021) Development and validation of epigenetic signature predict survival for patients with laryngeal squamous cell carcinoma. DNA Cell Biol 40(2):247–264
Metzger K et al (2021) A six-gene expression signature related to angiolymphatic invasion is associated with poor survival in laryngeal squamous cell carcinoma. Eur Arch Otorhinolaryngol 278(4):1199–1207
Ji M, Zhang LJ (2021) Expression levels of SCCA and CYFRA 21–1 in serum of patients with laryngeal squamous cell carcinoma and their correlation with tumorigenesis and progression. Clin Transl Oncol 23(2):289–295
Salzman J (2016) Circular RNA expression: its potential regulation and function. Trends Genet 32(5):309–316
Zhong Y et al (2018) Circular RNAs function as ceRNAs to regulate and control human cancer progression. Mol Cancer 17(1):79
Yang G, Zhang Y, Yang J (2019) Identification of potentially functional CircRNA-miRNA-mRNA regulatory network in gastric carcinoma using bioinformatics analysis. Med Sci Monit 25:8777–8796
Zhang HD et al (2018) CircRNA: a novel type of biomarker for cancer. Breast Cancer 25(1):1–7
Yin Y et al (2019) Emerging roles of circRNA in formation and progression of cancer. J Cancer 10(21):5015–5021
Shuang Y et al (2021) A novel circular RNA circPPFIA1 promotes laryngeal squamous cell carcinoma progression through sponging miR-340-3p and regulating ELK1 expression. Bioengineered 12(1):5220–5230
Wu T et al (2021) Hsa_circ_0042823 accelerates cancer progression via miR-877-5p/FOXM1 axis in laryngeal squamous cell carcinoma. Ann Med 53(1):960–970
Zheng Y et al (2021) Circ_0023028 contributes to the progression of laryngeal squamous cell carcinoma by upregulating LASP1 through miR-486-3p. Mol Cell Biochem 476(8):2951–2961
Han J, Lin Q, Dong C (2021) Plasma cell-free circRNAs panel act as fingerprint predicts the occurrence of laryngeal squamous cell carcinoma. Aging (albany NY) 13(13):17328–17336
Chen Q et al (2020) miR-145–5p suppresses proliferation, metastasis and EMT of colorectal cancer by targeting CDCA3. Pathol Res Pract 216(4):152872
Tang W et al (2019) miR-145-5p suppresses breast cancer progression by inhibiting SOX2. J Surg Res 236:278–287
Gao W et al (2019) Identification of miR-145–5p-centered competing endogenous RNA network in laryngeal squamous cell carcinoma. Proteomics 19(21–22):e1900020
Gao W et al (2019) Promoter methylation-regulated miR-145-5p inhibits laryngeal squamous cell carcinoma progression by targeting FSCN1. Mol Ther 27(2):365–379
Ding Y et al (2017) Analysis of gene expression profiling variations induced by hsamiR1455poverexpression in laryngeal squamous cell carcinoma cell line Tu177. Mol Med Rep 16(5):5863–5870
Huang B et al (2018) OTX1 regulates cell cycle progression of neural progenitors in the developing cerebral cortex. J Biol Chem 293(6):2137–2148
Zhang YF et al (2015) Otx1 promotes basal dendritic growth and regulates intrinsic electrophysiological and synaptic properties of layer V pyramidal neurons in mouse motor cortex. Neuroscience 285:139–154
Qin SC et al (2018) Dowregulation of OTX1 attenuates gastric cancer cell proliferation, migration and invasion. Oncol Rep 40(4):1907–1916
Chen S et al (2020b) MicroRNA-4516 suppresses pancreatic cancer development via negatively regulating orthodenticle homeobox 1. Int J Biol Sci 16(12):2159–2169
Li H et al (2016) OTX1 contributes to hepatocellular carcinoma progression by regulation of ERK/MAPK pathway. J Korean Med Sci 31(8):1215–1223
Tu XP et al (2020) OTX1 exerts an oncogenic role and is negatively regulated by miR129-5p in laryngeal squamous cell carcinoma. BMC Cancer 20(1):794
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Zhongshun He and Qiannan Xu designed and performed the research; Cailian Li and Biao Xu analyzed the data; Shihua Li and Ying Zhang wrote the manuscript. All authors read and approved the final manuscript.
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Written informed consents were obtained from all participants and this study was permitted by the Ethics Committee of Stomatological Hospital affiliated to Kunming Medical University.
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Shihua Li and Ying Zhang contributed equally to this work as co-first authors.
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10142_2022_862_Fig10_ESM.png
Supplementary Fig. 1 Detection of cell viability and screening of target genes. (A-B) Cell viability was detected by cell counting kit 8 assay in Tu177 and SNU899 cells transfected with sh-NC or sh-circMYOF. (C) The expression of candidate mRNA was detected by qRT-PCR in Tu177 cells transfected with miR-145-5p mimic or miR-NC. (D) Kaplan-Meier analysis was performed to analyze the relationship between OTX1 expression and the overall survival rate of LSCC patients.*p < 0.05. (PNG 65.0 KB)
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Li, S., Zhang, Y., He, Z. et al. Knockdown of circMYOF inhibits cell growth, metastasis, and glycolysis through miR-145-5p/OTX1 regulatory axis in laryngeal squamous cell carcinoma. Funct Integr Genomics 22, 1–13 (2022). https://doi.org/10.1007/s10142-022-00862-8
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DOI: https://doi.org/10.1007/s10142-022-00862-8