<p>Anti-Breast Cancer Effect of 2-Dodecyl-6-Methoxycyclohexa-2,5-Diene-1,4-Dione in vivo and in vitro Through MAPK Signaling Pathway</p>

Abstract

Background: 2-Dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione (DMDD) has been reported to inhibit a variety of cancer cell lines. The purpose of this study was to investigate the effects of DMDD on 4T1 breast cancer cells and the effects of DMDD on 4T1 breast cancer in mice and its molecular mechanisms. Methods: 4T1 breast cancer cells were treated with different concentrations of DMDD, and their proliferation, apoptosis, cell-cycle distribution, migration, and invasion were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT, Acridine orange and ethidium bromide dual staining analysis (AO/EB) dual staining, flow cytometry, scratch test, and the Transwell assay. Relative quantitative real-time qPCR analysis and Western blot were applied to examine the expression levels of related genes and proteins. In animal experiments, we established a xenograft model to assess the anti-breast cancer effects of DMDD by evaluating the inhibition rate. The apoptotic activity of DMDD was evaluated by hematoxylin-eosin (HE) staining, transmission electron microscope (TEM) analysis and TdT-mediated dUTP nick end labeling (TUNEL) assays. The mRNA expression levels of MAPK pathway components were detected by relative quantitative real-time qPCR. In addition, the protein expression levels of MAPK pathway components were assessed through immunohistochemical assays and Western blotting. Results: Experiments showed that DMDD could inhibit the proliferation, migration, invasion of 4T1 cells and induce cellular apoptosis and G1 cell cycle arrest. Moreover, DMDD down-regulated the mRNA expressions of raf1, mek1, mek2, erk1, erk2, bcl2, and upregulated the mRNA expression of bax. DMDD reduced the protein expressions of p-raf1, p-mek, perk , p-p38, Bcl2, MMP2, MMP9 and increased the protein expressions of Bax and p-JNK. The results showed that DMDD can effectively reduce the tumor volume and weight of breast cancer in vivo, up-regulate the expression of IL-2, down-regulate the expression of IL-4 and IL-10, induce the apoptosis of breast cancer cells in mice, and regulate the expression of genes and proteins of the MAPK pathway. Conclusion: Our study indicates that DMDD can inhibit proliferation, migration, and invasion and induces apoptosis and cell-cycle arrest of 4T1 breast cancer cells. Also, our findings indicate that DMDD induces the apoptosis of breast cancer cells and inhibits the growth in mice. Its mechanism may be related to the MAPK pathway.

Effects of 2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione on autophagy and the PI3K/AKT/mTOR signaling pathway in human cholangiocarcinoma QBC939 cells

“…2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione (DMDD) is a quinone extracted from the root of Populus tremula, with hypoglycemic, lipid-regulating, and inflammation-reducing effects. Recent studies have found that DMDD can inhibit the proliferation of breast, lung, and bone cancer cells, as well as inhibit the growth of breast cancer in mice (22)(23)(24). In this study, we investigated the effect of DMDD on human QBC939 CCA cells, and tentatively demonstrated that DMDD is a novel inducer of autophagy and induces autophagy by regulating the PI3K/AKT/mTOR signaling pathway in the QBC939 cell line.…”

Uncovering the Pharmacological Mechanism of 2-Dodecyl-6-Methoxycyclohexa-2,5 -Diene-1,4-Dione Against Lung Cancer Based on Network Pharmacology and Experimental Evaluation

“…In previous study, an active compound DMDD was separated from the roots of Averrhoa Carambola L. (Oxalidaceae) and observed to contain several biological properties ( Ouyang et al, 2014 ). Previous studies have suggested that DMDD shows significant antitumor potential against human breast cancer by inhibiting the TLR4/MyD88/NF-κB ( Lu et al, 2019 ) and MAPK pathway ( Zhou et al, 2020 ) in vitro ( Chen et al, 2017 ) and in vivo . However, the potential targets and pharmacological mechanism underlying the regulation of DMDD in lung cancer tumorigenesis still remain unclear.…”

“…The anticancer effects of natural products and their secondary metabolites are widely used in the treatment of tumors, including paclitaxel, vincristine, etoposide and camptothecin ( Ouyang et al, 2014 ). 2-dodecyl-6-methoxycyclohexa-2, 5-diene-1,4-dione (DMDD), an active quinonoids from the roots of Averrhoa carambola L. , exhibits various biological effects, including antidiabetic ( Kintoko et al, 2018 ), antidiabetic nephropathy ( Lu et al, 2019 ) and anti-breast cancer ( Gao et al, 2015 ; Chen et al, 2017 ; Zhou et al, 2020 ) activities. The various biological activities and low toxicity of DMDD make it a promising candidate for clinical development.…”

MicroRNA-497-5p stimulates osteoblast differentiation through HMGA2-mediated JNK signaling pathway

“…Pre-treatment with MAPK inhibitors reduced the protein expression of Bax promoted by IL-1α in the MC3T3-E1 cells, suggesting the significance of JNK and the p38 MAPK signaling in modulating IL-1α-induced apoptosis and osteoblast differentiation of MC3T3-E1 cells [ 30 ]. In addition, phosphorylation of JNK leads to an increase in apoptosis rate in cells under different conditions, such as ischemia-reperfusion [ 31 , 32 ], cancers [ 33 , 34 ], and liver injury [ 35 ]. Moreover, miR-214 inhibitor significantly decreased the expression of ALP, OCN, and RUNX2, as well as ALP activity in MC3T3-E1 cells, which was enhanced by additional treatment with SP600125, a JNK inhibitor [ 36 ].…”