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Archive > Volume 31 Issue 6 > 2024,31(6):592-597. DOI:10.3872/j.issn.1007-385X.2024.06.008 Prev Next
Basic Research
miR-218-5p regulates glycolysis in human non-small cell lung cancer A549 cells by targeting PDE7A
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Abstract:
Objective: To investigate the mechanism of miR-218-5p regulating the glycolytic process in human non-small cell lung cancer A549 cells by targeting phosphodiesterase 7A (PDE7A). Methods: A549 cells were routinely cultured, and miR-218-5p mimic, mimic-NC, PDE7A overexpression plasmid (PDE7A-oe) and PDE7A control plasmid (PDE7A-NC) were transfected into A549 cells using Lipo3000, and recorded as the miR-218-5p mimic group, the mimic-NC group, the PDE7A-oe group and the PDE7A-NC group. The transfection efficiency was verified by qPCR assay; the expressions of glycolysis key enzyme proteins were detected by WB assay; the 2-deoxyglucose and lactate contents in A549 cells of each transfection group were detected by glucose assay and lactate production assay; the target binding relationship between miR-218-5p and PDE7A was verified by dual-luciferase reporter gene assay, and the data from the TCGA database were used to analyze the expression level of PDE7A mRNA in lung cancer tissues. Results: miR-218-5p was successfully overexpressed in A549 cells (P<0.01). Overexpression of miR-218-5p significantly inhibited the expressions of PDE7A, HK2, PKM2 proteins (all P<0.01), glucose uptake and lactate production (both P<0.01) in A549 cells. Overexpression of PDE7A significantly promoted the expressions of PDE7A, HK2, and PKM2 proteins (all P<0.01), as well as glucose uptake and lactate production (both P<0.01) in A549 cells. miR-218-5p in A549 cells could directly bind to the 3′-UTR of PDE7A mRNA. Database data analysis showed that PDE7A mRNA was highly expressed in lung squamous cell carcinoma tissues (P<0.01). Conclusion: miR-218-5p targets PDE7A to regulate the expression levels of HK2 and PKM2 in A549 cells, which in turn inhibits the glycolytic process. miR-218-5p/PDE7A may be a potential target for clinical diagnosis and treatment of NSCLC.
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