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OBJECTIVE:To investigate the effect of Ruyanneixiao cream (RYNX) on the expression of hypoxia inducible factor-1α (HIF-1c),hexokinase 2 (HK2),phosphofructokinase (PFK),and pyruvate kinase M2 (PKM2) mRNA and protein in MCF-10AT cells and in an animal model of precancerous mammary lesions.METHODS:Following treatment of MCF-10AT cells with RYNX,tamoxifen (TAM) and YC-1 for 48 h,HIF-1α,HK2,PFK,PKM2 mRNA and protein expression was analyzed.Fifty female SD rats were randomly divided into control,model,TAM,and highand low-dose RYNX groups,with 10 rats in each group.A precancerous mammary lesion model was established for all groups except the control group.High-and low-dose RYNX cream containing TAM was coated on the breasts of animals in the corresponding groups.The rat mammary tissue was removed in the 10th week and HIF-1α,HK2,PFK,PKM2 mRNA and protein was analyzed.RESULTS:In vitro analyses demonstrated that,compared with the matrix group,HIF-1α,HK2,PFK,PKM2 mRNA and protein expression was significantly decreased in the RYNX group (P ＜ 0.05).Compared with the YC-1 + RYNX group,HK2,PFK,and PKM2 protein expression was significantly reduced in the RYNX group.HIF-1α,HK2,PFK,and PKM2 protein expression was increased significantly in the model group (P ＜ 0.05) compared with the control group,while HIF-1α,HK2,PFK,and PKM2 mRNA and protein expression was significantly decreased in both the high-and low-dose RYNX groups (P ＜ 0.05),with the effect being greater in the high-dose group.CONCLUSION:RYNX can block precancerous breast lesions by decreasing the expression of HK2,PFK,and PKM2 mRNA and protein via inhibition of HIF-1α mRNA and protein overexpression in a dose-dependent manner.