Circ_0000376 regulates miR-577/HK2/LDHA signaling pathway to promote the growth, invasion and glycolysis of osteosarcoma

• Published in: Journal of orthopaedic surgery and research Vol. 19; no. 1; p. 67
• Main Authors: Dai, Hongchun, Yi, Guangming, Jiang, Dong, Min, Yanmei, Li, Zongwei
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 13-01-2024; BioMed Central; BMC
• Subjects: Analysis; Animal models; Animals; Apoptosis; Bone cancer; Bone Neoplasms - genetics; Cell cycle; Cell growth; Cell Line, Tumor; Cell proliferation; Cell Proliferation - genetics; circ_0000376; Circular RNA; Development and progression; Dextrose; Flow cytometry; Glucose; Glucose metabolism; Glycolysis; Glycolysis - genetics; Growth; Hexokinase; Hexokinase - genetics; HK2; Humans; L-Lactate dehydrogenase; Lactic acid; LDHA; Luciferase; Metastasis; Mice; MicroRNA; MicroRNAs; MicroRNAs - genetics; miR-577; miRNA; Molecular modelling; Orthopedics; Osteosarcoma; Osteosarcoma - genetics; Proteins; Reagents; Sarcoma; Signal transduction; Signal Transduction - genetics; Software; Tumorigenesis; Massachusetts; China; Beijing China; United States > US; LDHA; miR-577; circ_0000376; Osteosarcoma; HK2
• ISSN: 1749-799X; 1749-799X
• DOI: 10.1186/s13018-023-04520-y

Many studies have confirmed that circular RNAs (circRNAs) mediate the malignant progression of various tumors including osteosarcoma (OS). Our study is to uncover novel molecular mechanisms by which circ_0000376 regulates OS progression.
The expression of circ_0000376, microRNA (miR)-577, hexokinase 2 (HK2) and lactate dehydrogenase-A (LDHA) was determined by quantitative real-time PCR. OS cell proliferation, apoptosis and invasion were measured using cell counting kit 8 assay, colony formation assay, EdU assay, flow cytometry and transwell assay. Besides, cell glycolysis was assessed by testing glucose consumption, lactate production, and ATP/ADP ratios. Protein expression was examined by western blot analysis. The interaction between miR-577 and circ_0000376 or HK2/LADA was verified by dual-luciferase reporter assay. The role of circ_0000376 on OS tumor growth was explored by constructing mice xenograft models.
Circ_0000376 had been found to be upregulated in OS tissues and cells. Functional experiments revealed that circ_0000376 interference hindered OS cell growth, invasion and glycolysis. Circ_0000376 sponged miR-577 to reduce its expression. In rescue experiments, miR-577 inhibitor abolished the regulation of circ_0000376 knockdown on OS cell functions. MiR-577 could target HK2 and LDHA in OS cells. MiR-577 suppressed OS cell growth, invasion and glycolysis, and these effects were reversed by HK2 and LDHA overexpression. Also, HK2 and LDHA expression could be regulated by circ_0000376. In vivo experiments showed that circ_0000376 knockdown inhibited OS tumorigenesis.
Circ_0000376 contributed to OS growth, invasion and glycolysis depending on the regulation of miR-577/HK2/LDHA axis, providing a potential target for OS treatment.