Abstract: Objective To investigate the role of miR-181c in glycolysis of cancer-associated fibroblasts (CAFs) and explore the
mechanism. Methods Human lung CAFs and normal fibroblasts (NFs), isolated from fresh human lung adenocarcinoma tissue
specimens by primary culture of tissue explants, were transfected with a miR -181c mimics, a miR-181c inhibitor, a siRNA
siRNA-HK2 or the vector HK2-vector via LipofectamineTM 2000. Quantitative real-time PCR was used to analyze the changes in
miR-125b expression in the transfected cells; hexokinase-2 (HK2) protein expression in the cells was detected using Western
blotting, and the cellular glucose uptake was assessed with 2-NBDG. Lactate production in the cells was examined and
expression of HK2 mRNA was detected with dual luciferase reporter gene assay. Results No obvious difference was found in
the cell morphology between CAFs and NFs. Compared with the NFs, the CAFs showed obviously increased glucose uptake,
lactate production and HK2 protein expression with decreased expressions of the miR-181 family (P<0.05). Transfection with
the miR-181 inhibito- rsignificantly increased glucose uptake, lactate production and HK2 protein expression in the NFs. In
CAFs, transfection with the miR-181 mimics caused significantly lowered glucose uptake, lactate production and HK2 protein
expression of. Knockdown of endogenous HK2 by siRNA abolished miR-181 mimics-mediated decrease of glucose uptake and
lactate production in CAFs, while transfection with miR-181 mimics suppressed HK2 overexpression-induced enhancement of
glucose uptake and lactate production in NFs. Conclusion Transfection with miR-181 mimics can suppress glycolysis in CAFs
by inhibiting HK2 expression.