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

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 up-regulated the mRNA expression of bax. DMDD reduced the protein expressions of p-raf1, p-mek, p-erk, 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.

Protective Effect of Benzoquinone Isolated from the Roots of Averrhoa carambola L. on Streptozotocin-Induced Diabetic Mice by Inhibiting the TLR4/NF-κB Signaling Pathway.

BACKGROUND: Studies have demonstrated that the roots of Averrhoa carambola L. (Oxalidaceae), a traditional Chinese medicine, can be used to treat diabetes and diabetes-related diseases. Nevertheless, the potential beneficial effects and mechanism of benzoquinone isolated from the roots of Averrhoa carambola L. (BACR) on diabetes remain unclear. METHODS: Diabetic Kunming mice were injected with STZ (120 mgkg-1) in the tail vein. Fasting blood glucose (FBG) and the change of body weight were measured after oral administration of BACR (120, 60, 30 mg/kg/d) every week. The levels of the total cholesterol (TC), triglyceride (TG), free fatty acids (FFA), glucosylated hemoglobin (GHb), fasting insulin (FINS), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were measured. The histological examination of pancreatic tissues and the TLR4/NF-κB pathway was analyzed by RT-PCR, immunohistochemistry and Western blot. RESULTS: The study found that clearly the BACR obviously reduced the blood glucose, serum lipids, GHb and FINS. In addition, BACR treatment markedly reduced the release of inflammatory factors, including IL-6 and TNF-α, and down-regulated the expression of the TLR4/NF-κB pathway. CONCLUSION: BACR has potential benefits for the treatment of diabetes by ameliorating metabolic functions and attenuating the inflammatory response via inhibition of the activation of theTLR4/NF-κB pathway.

[Effect of enhanced UV-B radiation on polyamine metabolism and endogenous hormone contents in rice (Oryza sativa L.)].

The results showed that the activities of Agrinine decarboxylase(ADC), Ornithine decarboxylase(ODC) and s-Adenosylmethionine decarboxylase(SAMDC) were increased by 165.74%, 104.60% and 89.60% in the leaves of Shan You63(Sy63) and by 59.91%, 41.30% and 23.68% in the leaves of Nancheum(NC). Only ADC and ODC activities were increase by 115.93%, 14.45%, but SAMDC activity was decreased by 33.01% in the leaves of IR65600-85 respectively in the exposure to enhanced UV-B radiation for 7-14 days. In late treatment time course(21-28 d), the activities of ADC and ODC were increased by 89.72% and 3.71% in the leaves of Sy63 exposed to UV-B radiation for 21-28 days and by 73.95% and 27.38% in the leaves of NC. The activity of ADC was also increased by 94.41%, but ODC activity was decreased by 13.57% in the leaves of IR65600-85 compared with the controls. As far as SAMDC was concerned, the enzymic activities in the leaves of Sy63, NC and IR65600-85 were reduced by 40.06%, 19.20% and 38.21% respectively in the exposure to enhanced UV-B radiation for 21-28 days. The reverse was true in the case of Polyamine Oxidase (PAO), this in turn resulted in increased contents of Polyamine(PA) especially putrescine(Put). In addition, the result also indicated that the contents of IAA and GA1/3 were significantly reduced in all rice cultivars used for this experiments with enhanced UV-B radiation treatments for 7-28 days, in which the contents of IAA and GA1/3 were decreased by 58.92% and 45.48% in the leaves of Sy63, by 43.31% and 56.20% in the leaves of NC, and by 38.60% and 47.33% in the leaves of IR65600-85. The contents of ZRs in the leaves of the three cultivars concerned were lower in earlier treatment time courses (7-14 d), but much higher in late courses(21-28 d) compared with the their counterparts. With regard to the endogenous hormone of ABA, the content was significantly increased by 14.4%, 99.6% and 56.7% respectively in the three rice cultivars concerned exposed to enhanced UV-B radiation for 7-28 days, thereby led to decreased values of IAA/ABA, GA1/3/ABA and ZRs/ABA, consequently suppressed growth and development of rice.

[Effects of enhanced ultraviolet-B radiation on growth development and yield formation in rice (Oryza sativa L.)].

Effect of enhanced ultraviolet-B radiation (280-320 nm) on growth, development and yield formation in the three rice cultivars (Oryza sativa L.) under pot conditions were investigated. The results showed that enhanced ultraviolet-B radiation decreased rice height, tiller number, total leaf area and biomass, but the inhibitory rate varied in different cultivars and developmental stages. Plant height was decreased by 9.4%-12.2% in seedling stage, plant shoot and root of dry matter were decreased by 45.3%-59.8% and 54.9%-59.0% respectively in tillering period. The leaf emergence delayed, resulting in delayed time of flowering in Shanyou 63, Nanchuan and IR65600-85 by 2d, 3d and 7d, prolonged total growth period by 3d, 4d and 9d. Net photosynthesis rate of Shanyou 63, Nanchuan and IR65600-85 were decreased by 11.9%, 12.8% and 29.7%, respectively. Attributed to decreased contents of chlorophyll a, b, carotenoid and induction kinetics parameters of Fv, Fv/Fm and Fv/Fo. Effective panicle per plant, total grain number per panicle, filled grain rate and 1000-grain weight were decreased by enhanced UV-B radiation. Consequently the grain yields were reduced by 25.2%-31.1%.