External use of Ruyanneixiao cream efficiently blocks precancerous mammary lesions by interfering with glycolysis induced by inhibition of hypoxia inducible factor-1α, hexokinase 2, phosphofructokinase, and pyruvate kinase M2 expression

Abstract

ObjectiveTo investigate the effect of Ruyanneixiao cream (RYNX) on the expression of hypoxia inducible factor-1α (HIF-1α), 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. MethodsFollowing 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 high- and 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. ResultsIn 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. ConclusionRYNX 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.