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Effect of lncRNA MAFG-AS1 regulating miR-532-3p expression on glycolysis of lung cancer A549 cells
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Abstract:
Objective: To investigate the effect of lncRNA MAFG antisense RNA1 (lncRNA MAFG-AS1) regulating miR-532-3p on the glycolysis of lung cancer A549 cells. Methods: The expression levels of MAFG-AS1 and miR-532-3p in human lung cancer A549 cells and normal lung epithelial BEAS-2B cells were detected by qPCR. A549 cells with MAFG-AS1 downregulation or miR-532-3p overexpression were constructed by liposome transfection technique, respectively. The glucose consumption and lactate secretion in cell culture supernatant of A549 cells were detected by visible spectrophotometry, and the mRNA and protein expression levels of pyruvate kinase M2 (PKM2) and hexokinase 2 (HK2) in A549 cells were detected using qPCR and WB, respectively. The targeting relationship between MAFG-AS1 and miR-532-3p was verified by Dual luciferase reporter gene assay, and the effects of co-downregulation of MAFG-AS1 and miR-532-3p on glucose uptake, lactate secretion and expression of PKM2 and HK2 in A549 cells were observed. Results: Compared with BEAS-2B cells, the expression of MAFG-AS1 was upregulated while miR-532-3p expression was downregulated in A549 cells (all P<0.01). Glucose consumption, lactate secretion and the protein and mRNA expressions of PKM2 and HK2 were inhibited by MAFG-AS1 downregulation or miR-532-3p overexpression (all P<0.01). miR-532-3p could bind to MAFG-AS1 and inhibit the luciferase activity of wild-type MAFG-AS1 cells (P<0.01). Downregulation of MAFG-AS1 could increase the expression of miR-532-3p (P<0.01)，while the co-downregulation of miR-532-3p could reverse the effect of MAFG-AS1 downregulation on glucose uptake, lactate secretion and the expression of PKM2 and HK2 in A549 cells (all P<0.01). Conclusion: Downregulation of MAFG-AS1 can inhibit the glycolysis of A549 cells by promoting the expression of miR-532-3p.
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Project Supported:
Project supported by the Joint Construction Project of Medical Science and Technology Project of Henan Province (No.2018020557)
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