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
Author: Li XB, Ma M, Zhang GJ, Ma Y, Liao R, Chen RX, Yan XX, Bie FJ, Huang MJ, Liang SJ
OBJECTIVE: To investigate the effect of Ruyanneixiao cream (RYNX) on the expression of hypoxia inducible factor-1a (HIF-1a), 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-1a, 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-1a, HK2, PFK, PKM2 mRNA and protein was analyzed. RESULTS: In vitro analyses demonstrated that, compared with the matrix group, HIF-1a, 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-1a, HK2, PFK, and PKM2 protein expression was increased significantly in the model group (P<0.05) compared with the control group, while HIF-1a, 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-1a mRNA and protein overexpression in a dose-dependent manner.
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