Qing-Re-Yi-Liu decoction suppresses the malignant behaviors of breast cancer cells by attenuating the MnSOD/CaMKII/AMPK signaling and Warburg effect

Abstract

Background The Qing-Re-Yi-Liu decoction (QRYLD) is a clinical effective prescription for the treatment of breast cancer due to its activity of heat clearing and detoxification. Our preliminary studies have found that QRYLD can interfere with the Warburg effect of breast cancer cells, inhibiting the proliferation of breast cancer MCF-7 cells.The chemical components and molecular mechanisms underlying the actions of QRYLD in regulating the Warburg effect in breast cancer cells are still unclear. Methods The bioactive components of QRYLD aqueous extracts were detected by HPLC. The biological processes and signaling pathways in MCF-7 cells of QRYLD targets were measured with transcriptome analysis. The effect of QRYLD on the malignant behaviors of MCF-7 cells were analyzed by CCK-8 assay,transwell invasion assay, wound healing assay, apoptosis detection. The effect of QRYLD on glucose uptake, lactic acid production and Warburg effect in MCF-7 cells assessed by colorimetry and western blotting. The volumes of xenograft breast tumors and body weights of mice were measured, and the effect of QRYLD on the tumor tissues was assessed with immunohistochemistry. Results Here, we show that the QRYLD aqueous extracts contain several bioactive components. Analysis of transcriptomes indicated that QRYLD treatment altered the expression of many genes, such as manganese superoxide dismutase (MnSOD), that were involved in biological processes and signaling pathways, particularly for glucose metabolism in MCF-7 cells. Functionally, QRYLD treatment, like MnSOD silencing, inhibited the malignant behaviors of MCF-7 and enhanced their apoptosis while MnSOD over-expression had opposite effects. Furthermore, QRYLD treatment, like MnSOD silencing, limited glucose uptake and lactic acid production in MCF-7 cells, which were associated with a decrease in the relative levels of Glut-1, HIF-1α, c-Myc, HK-2, PFK-1, LDH-A, PKM-2, MnSOD, calmodulin dependent kinase II (CaMKII) and AMPK expression. Finally, treatment with QRYLD, like MnSOD silencing, significantly mitigated the growth of xenograft MCF-7 tumors in mice and reduced the expression of MnSOD, CaMkII and AMPK expression in the tumors. Conclusion These data suggest that QRYLD may target MnSOD to attenuate the MnSOD/CaMKII/AMPK signaling, leading to inhibition of the Warburg effect and malignant behaviors in MCF-7 cells. These findings may provide new insights into the pharmacological mechanisms underlying the actions of QRYLD in inhibiting the Warburg effect and malignant behaviors of breast cancer cells.