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Breast cancer is one of the leading causes of cancer-related deaths in women worldwide.It is a cancer that originates from the mammary ducts and involves mutations in multiple genes.Recently,the treatment of breast cancer has become increasingly challenging owing to the increase in tumor het-erogeneity and aggressiveness,which gives rise to therapeutic resistance.Epidemiological,population-based,and hospital-based case-control studies have demonstrated an association between high intake of certain Allium vegetables and a reduced risk in the development of breast cancer.Diallyl disulfide(DADS)and diallyl trisulfide(DATS)are the main allyl sulfur compounds present in garlic,and are known to exhibit anticancer activity as they interfere with breast cancer cell proliferation,tumor metastasis,and angiogenesis.The present review highlights multidrug resistance mechanisms and their signaling pathways in breast cancer.This review discusses the potential anticancer activities of DADS and DATS,with emphasis on drug resistance in triple-negative breast cancer(TNBC).Understanding the anticancer activities of DADS and DATS provides insights into their potential in targeting drug resistance mecha-nisms of TNBC,especially in clinical studies.
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Objective:To investigate the effect of diallyl trisulfide (DATS) on the expression of folate receptor α (FRα) in human gastric cancer cells and the related regulatory mechanism.Methods:Human gastric cancer cell lines BGC823 and AGS were selected, BGC823 cells were treated with 10, 20, 40, 80 μmol/L DATS for 48 hours, and AGS cells were treated with 5, 10, 20, 40 μmol/L DATS for 48 hours. Cells treated with 6 μmol/L histone deacetylase (HDAC) inhibitor trichostatin A (TSA) were used as positive control for epigenetic study, and cells untreated with DATS were used as negative control. The apoptosis of gastric cancer cells induced by DATS was detected by flow cytometry. After BGC823 and AGS cells were treated by DATS for 48 hours, they were replaced with DATS-free cell culture medium and cultured for different time to detect the changes in FRα protein expression. BGC823 and AGS cells were treated with 6 μmol/L TSA or 40 μmol/L DATS. The protein expression levels of FRα, HDAC1 and HDAC2, and histone H3 lysine 9 acetylation (H3K9ac) and histone H4 lysine 5 acetylation (H4K5ac) were detected by Western blot. BGC823 cells were inoculated into BALB/c nude mice to establish tumor bearing model. The nude mice in DATS group were injected intraperitoneally with 10 mg/kg DATS for 16 days, and then the protein in tumor-bearing tissues was extracted to detect the expression of target protein, while the control group was injected with equal dose of 0.9% NaCl solution.Results:The expressions of FRα protein in BGC823 and AGS cells were up-regulated in a dose-dependent way after gradient concentrations of DATS treatment ( F = 65.68, P < 0.01; F = 26.65, P < 0.01). After changing the cell culture medium without DATS, the expressions of FRα protein in BGC823 and AGS cells gradually decreased and returned to the initial levels ( F = 74.57, P < 0.01; F = 30.92, P < 0.01). With the increase of DATS concentration, the apoptosis rates of BGC823 and AGS cells increased ( F = 32.95, P < 0.01; F = 38.97, P < 0.01). After TSA treatment, FRα protein expressions in BGC823 and AGS cells were up-regulated by 4.5 times ( t = -12.62, P < 0.01) and 3.6 times ( t = -10.00, P < 0.01). After 40 μmol/L and 20 μmol/L DATS treatment, the expression level of FRα protein in BGC823 and AGS cells was up-regulated (both P < 0.01), the expressions of HDAC1 and HDAC2 were inhibited (all P < 0.01), and the levels of H3K9ac and H4K5ac acetylation modification increased (all P < 0.01). The results of tumor-bearing nude mice experiment showed that the volume of transplanted tumor in DATS group was smaller than that in the control group [(214±39) mm 3 vs. (577±98) mm 3], and the difference was statistically significant ( t = 4.86, P < 0.01). Compared with the control group, FRα protein expression in the transplanted tumor tissues of DATS group was up-regulated about 2 times ( t = -5.29, P < 0.01), and the expression levels of HDAC1 and HDAC2 proteins were down-regulated ( t = 9.36, P < 0.01; t = 9.88, P < 0.01). Conclusions:DATS up-regulates the expression of FRα protein in human gastric cancer BGC823 and AGS cells in a dose-dependent and reversible manner. The mechanism may be related to the effect of DATS on histone acetylation modification in tumor cells.
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Objective To study the proliferation and inflammatory phenotypes of human fibroblast-like synoviocytes (FLS) induced by tumor necrosis factor-α(TNF-α). Methods The rheumatoid arthritis (RA)-FLS were cultured in vitro, then treated with different concentrations of diallyl trisulfide (DATS). The proliferation activity was detected by CCK-8 method. Then TNF-α was used to stimulate the RA-FLS, mRNA and protein expression of interleukin (IL)-6, matrix metalloproteinases (MMP)-1 and vascular endothelial growth factor (VEGF) were detected by quantitative real time polymerase chain reaction (qPCR) and enzyme linked immunosorbent assay (ELISA). Differences among groups were determined by one-way analysis of variance (ANOVA), LSD-t test was used for comparison between 2 groups. Results RA-FLS was successfully is-olated and cultured in vitro. The positive rate of CD90 and CD29 in the RA-FLS was more than 90％. The proli-feration activity of RA-FLS treated with 100 μmoL/L, 200 μmol/L and 300 μmol/L DATS was (98.92 ± 0.40)％, (95.91±0.32)％, (94.05±0.24)％, respectively. As Compared with the normal control group, the pro-liferation activity of RA-FLS was lower, and the statistically significant difference is between normal control group 200 μmol/L and 300 μmol/L DATS (t=-4.46, P<0.05; t=-7.98, P<0.05). After TNF-α stimulation, the expression of IL-6's mRNA in experiment group (100μmol/L DATS) is higher compared with the model control group (t=5.74, P<0.05), but the change of IL-6's protein is no significant difference (t=-0.49, P=0.627). The differences of mRNA and protein expression levels of MMP-1 and VEGF between the experiment group (100μmol/L DATS) and the model control group were not statistically significant. The relative mRNA level [(0.42 ± 0.06), t=-23.47, P<0.05;(0.14±0.039), t=-36.59, P<0.05;(0.36±0.09), t=-13.1, P<0.05)] and the protein levels [(108.0±4.7) ng/L, t=-63.79, P<0.05, (26.0±1.0) ng/L, t=-9.68, P<0.05;(57.9±0.7), t=-34.59, P<0.05] of IL-6, MMP-1, VEGF in experiment group (200μmol/L DATS) were significantly decreased. And the relative mRNA level [(0.041 ±0.027), t=-38.48, P<0.05; (0.027 ±0.027), t=-41.22, P<0.05; (0.131 ±0.047), t=-17.74, P<0.05] and the protein levels [(24.2 ±2.3) ng/L , t=-88.69, P<0.05; (22.7 ±1.0) ng/L , t=-14.13, P<0.05; (34.5 ±1.7), t=-48.45, P<0.05] of IL-6, MMP-1, VEGF in the experiment group (300 μmol/L DATS) were also significantly decreased. The difference between the two groups was significant (t=-24.89, P<0.05; t=-4.45, P<0.05; t=-13.87, P<0.05). Conclusion DATS can inhibit the proliferation and the effect of TNF-αinduced secretion of IL-6, MMP-1 and VEGF in RA-FLS. The effect is dose-dependent.
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Objective To prepare diallyl trisulfide microemulsion (DATS-MEs) and evaluate its effect on the proliferation and migration of tumor cells. Methods Using HS15 as emulsifier, PEG400 as co-emulsifier and DATS as the oil phase, water titration method was used to prepare DATS-MEs. The influence of DATS-MEs on the proliferation of mouse Lewis cancer LLC cells and mouse melanoma B16BL6 cells in vitro was evaluated by the tetramethyl azoline blue (MTT) methed; Wound healing assay was used to evaluate the effect of DATS-MEs on the migration of LLC and B16BL6 cells in vitro. Results The DATS-MEs obtained was stable, clear, transparent, and well dispersed. The particle size of DATS-MEs was (16.72 ± 0.22) nm, the polydispersity index was 0.03 ± 0.01 and the Zeta potential was (-4.98 ± 0.11) mV. The encapsulation efficiency (EE) of DATS in microemulsion was 82.89%, the drug loading (DL) was 6.28%. The solubility in water increased by about 2 367 fold compared with the free DATS. DATS-MEs and free DATS at concentration of 10, 30, and 50 μmol/L all can effectively inhibit the migration and proliferation of LLC cells and B16BL6 cells in a dose-dependent manner. DATS microemulsion was proved to be more effective than free DATS. Conclusion After assembly into DATS microemulsion, the stability and solubility in water of DATS was enhanced. The anti-proliferation and anti-migration on mouse Lewis LLC cells and mouse melanoma B16BL6 cells were significantly enhanced.
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BACKGROUND: Diallyl trisulfide (DATS), a garlic-derived organosulfuric compound, has been documented for potential anti-inflammatory effects. However, the mechanism in microglia remains unknown. In this study, we investigated the anti-inflammatory effects of DATS in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. METHODS: The effects of DATS on LPS-induced pro-inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2) were assessed under conditions not in the cytotoxicity of DATS. The protein expression of inflammation regulatory genes was measured by Western blot analysis. RESULTS: DATS significantly inhibited the LPS-induced secretion of NO and PGE2, which was associated with the suppression of their regulatory genes, inducible NO synthase and COX-2. DATS had been shown to inhibit nuclear translocation of NF-κB by destroying the degradation and phosphorylation of IκB-α inhibitors in the cytoplasm. In addition, DATS effectively inhibited the expression of LPS-induced toll-like receptor 4 (TLR4) and myeloid differentiation factor 88. Furthermore, DATS markedly reduced the LPS-induced expression of chemokine (CXC motif) ligand (CXCL) 12 and CXC receptor (CXCR) 4, demonstrating its capacity to block chemo-attractive activity. CONCLUSIONS: These results indicate that DATS inhibits the activation of the CXCL12/CXCR4 axis associated with antagonizing effect on TLR4 and blocks NF-κB signaling, thus demonstrating anti-inflammatory effects against LPS stimulation.
Subject(s)
Blotting, Western , Cytoplasm , Dinoprostone , Genes, Regulator , Inflammation , Microglia , Myeloid Differentiation Factor 88 , Nitric Oxide , Nitric Oxide Synthase , Phosphorylation , Toll-Like Receptor 4 , Toll-Like ReceptorsABSTRACT
AIM:To study the effect of diallyl trisulfide (DATS) on experimental corneal neovascularization (CNV) in rats induced by corneal suture and detect the expression of vascular endothelial growth factor (VEGF) and p-AKT in rats cornea.METHODS:The rat model of corneal neovascularization (CNV) was induced by corneal suture.Rats were randomly divided into Group A:physiological saline control group containing DMSO (10 rats);Group B:25μmol/L DATS treatment group (10 rats);Group C:50μmol/L DATS treatment group (10 rats);Group D:100μmol/L DATS treatment group (10 rats);Group E:200μmol/L DATS treatment group (10 rats).The occurrence and development of CNV were observed by slit-lamp microscope at 7d after suture,and the area of CNV were calculated.Two weeks later,HE staining was used to observe the pathological organization form of each cornea,and RT-PCR and Western blot were used to detect the expression of VEGF mRNA and protein expression of VEGF and p-AKT between each groups.RESULTS:The blood vessel area of Group C,D and E was compared with that of Group A,the difference was statistically significant (P＜0.05);HE slice showed corneal edema,angiogenesis and inflammation infiltration situation gradually reduced comparing with the Group A,with the increase of concentration of DATS.RT-PCR showed the expression of VEGF mRNA in Group B,C,D,and E decreased compared with the Group A,and the difference was statistically significant (P＜0.05).Western-blot showed that the expressions of VEGF and p-AKT in Group B,C,D and E decreased gradually compared with those in Group A,and the difference was statistically significant (P＜0.05).CONCLUSION:DATS can inhibit corneal neovascularization of the rats induced by suture.Its mechanism may be associated with suppression of VEGF secretion,down-regulation of VEGF and inactivation of p-AKT.
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Objective: To investigate the effects of DATS (diallyl trisulfide) on cell proliferation, cell cycle and apoptosis of human osteosarcoma cell line Saos-2, and to explore their potential molecular mechanisms. Methods: Saos-2 cells were treated with 25, 50 and 100 μmol/L DATS for 24, 48, 72, and 96 h. The change of cell morphology was observed under an inverted microscope. The effect of DATS on the proliferation of Saos-2 cells was detected by MTT assay. After treatment with DATS at desired concentrations for various time intervals, the cell cycle and apoptotic rate of Saos-2 cells were analyzed by flow cytometry. The expressions of DATS-sensitive proteins in Saos-2 cells were systematically detected using 2-DE (two-dimensional gel electrophoresis) and mass spectrometry analysis. Results: DATS could inhibit the proliferation of Saos-2 cells in a dose- and time-dependent manner. Moreover, DATS could arrest the cell cycle at phase G0/G1 and induce the apoptosis in a time- and concentrationdependent manner. A total of 27 unique DATS-sensitive proteins, including 18 downregulated proteins and 9 upregulated proteins, were found in Saos-2 cells. Conclusion: DATS can suppress the proliferation of Saos-2 cells by blocking cell cycle progression and inducing apoptosis. The results of proteomics not only suggest the anticarcinogenic mechanism of DATS, but also provide a novel insight into the therapeutic targets for gene therapy of osteosarcoma. Copyright © 2013 by TUMOR.
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Health benefits of garlic and other Allium vegetables (e.g., onions), such as lipid lowering and anticancer effects, are credited to metabolic byproducts, including diallyl trisulfide (DATS). Evidence for anticancer effects of garlic derives from both population-based case-control studies, and clinical and laboratory investigations using purified garlic constituents such as DATS. Studies have shown that DATS can offer protection against chemically-induced neoplasia as well as oncogene-driven spontaneous cancer development in experimental rodents. Mechanisms underlying cancer chemopreventive effects of DATS are not completely understood, but known pharmacological responses to this natural product include alteration in carcinogen-metabolizing enzymes, cell cycle arrest, induction of apoptotic cell death, suppression of oncogenic signal transduction pathways, and inhibition of neoangiogenesis. This article reviews mechanisms and targets of cancer chemoprevention by DATS.
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[Objective]This study was designed to determine growth inhibition of diallyl trisulfide(DATS)in human prostate cancer cells by inducing apoptosis and further to investigate the mechanism underlying such effect.[Methods]Growth inhibition by DATS was estimated by the tetrazolium(MTr)assay.Apoptosis induction in DATS-treated cells was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with DAPI and flow cytometric analysis of cells with sub-G1 DNA content following staining with propidium iodide.Protein levels of apoptosis regulating proteins were determined using western blot.The activity of caspase-3 was measured using a colorimetric assay.[Result]DATS showed tumor growth inhibition in a time-and dose-dependent manner,IC_(50) of DATS was 14 μmol/L at 72 h.DATS evoked apoptosis as confirmed by cell morphology and by the analysis of flow cytometry.The expression of Bcl-2 and Bcl-xL,the apoptosis-suppressing proteins,was more down-regulated.The activity of caspase-3 was enhanced by DATS.[Conclusion]DATS inhibits growth of prostate cancer cells by inducing apoptosis in association with down-regulation of Bcl-2 and Bcl-xL and activation of caspase-3.
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Objective To investigate the role of nuclear factor-kappa B(NF-?B)in the modulation of diallyl trisulfide(DATS)on interleukin-1?(IL-1?)expression induced by lipopolysaccharide(LPS)in mice with acute lung injury(ALI).Methods Mice were randomly divided into Control group,ALI group,DATS group,DATS prevention group and DATS treatment group.The expression of IL-1? mRNA in the lung tissue was detected by reverse transcription PCR(RT-PCR).NF-?B activity in the lung tissue was detected by electrophoresis mobility shift assay(EMSA).The expression of phospho-I?B and I?B were assayed by Western blot.Results The expression of IL-1? mRNA,NF-?B activity and the phospho-I?B expression in lung tissues increased significantly at ALI group(P
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Aim To investigate whether DATS induce MGC803 cell apoptosis and the relationship betweenapoptosis and Ca~(2+) disruption. Methods MGC-803 cell growth inhibition was measured by MTT assay. Tunnel and flow cytometry methods were used to determine the induction of apoptosis and Ca2+ homeostasis disruption. Result MTT assay showed that the inhibitory rates on MGC-803 cell growth of different concentrations of DATS 4,8,12,16 and 24 mg?L-1 were 0.231?0.037,0.305?0.036,0.455?0.029,0.607?0.058,0.751?0.019 respectively. Flow cytometry analysis showed that treating MGC803 cell with DATS significantly increased the percentage of apoptosis cells and intracellular Ca2+. Treatment of cells with 1,2-bis(2-aminophenoxye-thane)-N,N,N-tetraacetic acid tetrakis acetoxymethyl ester (BAPTA-AM), cellular Ca2+ chelator, resulted in abolishment of the elevation of intracellular Ca~(2+) and blockage of DATS induced apoptotic of MGC-803. Conclusoin DATS could induce apoptosis of MGC-803 cells through the mechanism of Ca~(2+) homeostasis disruption.
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AIM: To study the effects of growth inhibition of different concentrations of diallyl trisulfide(DATS) on gastric cancer MGC-803 cell line in vitro. METHODS: The influence of different concentrations of DATS were examined by MTT assay, clonal formation rates and cell growth curve. RESULTS: Suppression and decrease of MGC-803 cell proliferation was found after treatment by DATS in vitro. The inhibitory rates on MGC-803 cell growth of different concentration of DATS, 4, 8, 12, 16 and 24 mg·L-1, were 26%, 46%, 65%, 76% and 89% respectively, and its half inhibitory concentration (IC50) was 8.2 mg·L-1. The clonal formation rates and clonal formation relative counts of 8, 12, 16 and 24 mg·L-1 were 32.4% and 58.7%, 24.8% and 42.5%, 19.0% and 33.5%, 8.8% and 15.1% respectively. There was significant correlation between dose and effect in all, and the cell growth culve became lower and flatter when concentration of DATS increase gradually. CONCLUSION: The effect of growth inhibition of DATS for gastric cancer MGC-803 cell in vitro is remarkable.
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OBJECTIVE:To investigate the stability of garlic oil-hydropropyl-?-cyclodextrin(HP-?-CD)inclusion complex.METHODS:With diallyl trisulfide(DATS)as determination index,the water solution of garlic oil HP-?-CD in-clusion complexes,freeze-drying powders of inclusion complex and the garlic oil injections were respectively subjected to il-lumination,high temperature,dilution tests and accelerated tests,and long-term sample observation tests by gas chro-matography(GC).RESULTS:The content of DATS in freeze-drying powders of inclusion complex had a minimum reduc-tion among the three under conditions of illumination and high temperature.In the accelerated tests and long-term sample observation tests,almost all the determination indexes of freeze-drying powders remained unchanged and a good stability was noted.Within10h,all the different diluents showed no significant effects on the indexes of the inclusion complex.CON-CLUSIONS:As compared with garlic oil injections,the garlic oil inclusion complex had a higher stability,freeze-drying powders of inclusion complex showed the best stability,yet the storing of which still needed to be under a proper temperature and away from light.
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AIM:To explore the production and cytotoxicity of the reactive oxygen species(ROS)induced by diallyl trisulfide(DATS)in HL-60 cells.METHODS:HL-60 cells were either treated with various doses of DATS alone,or DATS combination with apocynin,a specific NADPH oxidase inhibitor,or with antioxidant N-acetyl-L-cysteine(NAC)for 0,1,3,6,12 and 24 h,respectively.The intracellular ROS level was measured by flow cytometry.The activity of NADPH oxidase was evaluated by NBT reduction experiment.The content of both malondialdehyde(MDA)and the protein carbonyl were analyzed by spectrophotometer.RESULTS:The results from flow cytometry indicated that DATS significantly increased the intracellular ROS level in HL-60 cells(P
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Objective: To study the affection of Diallyl trisulfide on cultured human colon carcinoma cell line HT-29 and its mechanism. Methods: The viability of cells after various treatments were determined using the method of MTT. Flow cytometry were used to analyze the change of cell cycle and Bcl-2, Bax levels. Results: The proliferation of human colon carcinoma HT-29 cells were inhibited by Diallyl trisulfide. The inhibition was associated with dose and time dependence. Flow cytometry results showed that: 1) When cells were treated with Diallyl trisulfide at the concentration of 10 ?g/mL for 12 h and 24 h, the percentage of G0/G1 phase cells was decreased and that of G2/M phase cells was significantly increased compared with those in the control group.2) Diallyl trisulfide down regulated Bcl expression, while up-regulated bax expression. Conclusions: It was suggested that Diallyl trisulfide inhibited HT29 cell proliferation. And these were associated with arrest of cell cycles and down regulation of Bcl2/Bax ratio.
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AIM To study the effects of growth inhibition of different concentrations of diallyl trisulfide(DATS) on gastric cancer MGC 803 cell line in vitro. METHODS The influence of different concentrations of DATS were examined by MTT assay, clonal formation rates and cell growth curve. RESULTS Suppression and decrease of MGC 803 cell proliferation was found after treatment by DATS in vitro. The inhibitory rates on MGC 803 cell growth of different concentration of DATS,4, 8, 12, 16 and 24 mg?L -1 , were 26%,46%,65%,76% and 89% respectively, and its half inhibitory concentration (IC 50 ) was 8 2 mg?L -1 . The clonal formation rates and clonal formation relative counts of 8, 12, 16 and 24 mg?L -1 were 32 4% and 58 7%,24 8% and 42 5%,19 0% and 33 5%?8 8% and 15 1% respectively.There was significant correlation between dose and effect in all, and the cell growth culve became lower and flatter when concentration of DATS increase gradually. CONCLUSION The effect of growth inhibition of DATS for gastric cancer MGC 803 cell in vitro is remarkable.
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Aim To investigate the differential expression of apoptosis-associated genes in human gastric cancer MGC803 cells induced by diallyl trisulfide(DATS).Methods Growth inhibition against MGC803 cells was assayed by MTT assay;The apoptosis induced by DATS was assessed by Flow cytometry and fluorescent microscope.The apoptosis-associated gene expression of MGC803 cell treated with DATS was determided by Human Apoptosis Gene Array.Apaf-1 and SODD genes were confirmed by RT-PCR.Results DATS had significant growth inhibitory activity against MGC803 cells,inhibition ratio increased from 11% to 78% at 4,8,12,16 and 24 mg?L-1 for 72 h(P