microR-1294-5p inhibits glycolytic metabolism of non-small cell lung cancer cells via targeting TMPRSS11B

Non-smallcell lung cancer(NSCLC)cells intake and consume glucose at high efficiency by aerobic glycolysis to maintain robust cell growth and resist cell death.MicroRNAs(miRNAs)have been known to play pivotal roles in NSCLC development partly through mediating glycolysis.However,only a few miRNAs have been experimentally confirmed as critical regulators of glycolysis in NSCLC.TCGA datasets were analyzed to screen for differentially expressed miRNAs between NSCLC and normal tissues.The function of miR-1294-5p was determined in NSCLC cells by cell proliferation,glucose uptake,lactate release,and Extracellular Acidification Rate(ECAR)assays.The target of miR-1294-5p was predicted by TargetScan and miRDB,which was further validated by flow cytometry analysis,RT-qPCR,western blotting,a dual-luciferase reporter assay,and RNA immunoprecipitation(RIP)assay.In the present study,it was found that miR-1294-5p was a significantly downregulated miRNA in lung adenocarcinoma(LUAD)and lung squamous cell carcinoma(LUSC).The overexpression of miR-1294-5p inhibited glycolysis,lactate export,ECAR,and cell proliferation in NSCLC cells.Analysis with bioinformatic tools,Western Blotting,RT-qPCR,flow cytometry analysis,dual-luciferase reporter assay,and RIP assay showed that miR-1294-5p directly bound to complementary sites in the 3’-Untranslated Region(UTR)of TMPRSS11B resulted in downregulation of TMPRSS11B expression.In addition,transfection of recombinant TMPRSS11B rescued the functions of miR-1294-5p on glycolysis and proliferation of NSCLC cells.The findings provided novel insights for understanding the regulation of glycolytic metabolism in NSCLC.

miR-378-mediated glycolytic metabolism enriches the Pax7^(Hi) subpopulation of satellite cells

Adult skeletal muscle stem cells,also known satellite cells(SCs),are a highly heterogeneous population and reside between the basal lamina and the muscle fiber sarcolemma.Myofibers function as an immediate niche to support SC self-renewal and activation during muscle growth and regeneration.Herein,we demonstrate that microRNA 378(miR-378)regulates glycolytic metabolism in skeletal muscle fibers,as evidenced by analysis of myofiber-specific miR-378 transgenic mice(TG).Subsequently,we evaluate SC function and muscle regeneration using miR-378 TG mice.We demonstrate that miR-378 TG mice significantly attenuate muscle regeneration because of the delayed activation and differentiation of SCs.Furthermore,we show that the miR-378-mediated metabolic switch enriches Pax7^(Hi) SCs,accounting for impaired muscle regeneration in miR-378 TG mice.Mechanistically,our data suggest that miR-378 targets the Akt1/FoxO1 pathway,which contributes the enrichment of Pax7^(Hi) SCs in miR-378 TG mice.Together,our findings indicate that miR-378 is a target that links fiber metabolism to muscle stem cell heterogeneity and provide a genetic model to approve the metabolic niche role of myofibers in regulating muscle stem cell behavior and function.

Individual variation in buffalo somatic cell cloning efficiency is related to glycolytic metabolism

Mammalian individuals differ in their somatic cell cloning efficiency,but the mechanisms leading to this variation is poorly understood.Here we found that high cloning efficiency buffalo fetal fibroblasts(BFFs)displayed robust energy metabolism,looser chromatin structure,high H3 K9 acetylation and low heterochromatin protein 1α(HP1α)expression.High cloning efficiency BFFs had more H3 K9 ac regions near to the upstream of glycolysis genes by Ch IP-seq,and involved more openness loci related to glycolysis genes through ATAC-seq.The expression of these glycolysis genes was also found to be higher in high cloning efficiency BFFs by q RT-PCR.Two key enzymes of glycolysis,PDKs and LDH,were confirmed to be associated with histone acetylation and chromatin openness of BFFs.Treatment of low cloning efficiency BFFs with PS48(activator of PDK1)resulted in an increase in the intracellular lactate production and H3 K9 acetylation,decrease in histone deacetylase activity and HP1αexpression,less condensed chromatin structure and more cloning embryos developing to blastocysts.These results indicate that the cloning efficiency of buffalo somatic cells is associated with their glycolytic metabolism and chromatin structure,and can be improved by increasing glycolytic metabolism.