Of interest, our findings suggest that cell cycle perturbation and oxidative stress dominate at the low sulforaphane doses, whereas apoptosis and caspase activation dominate at the higher dose. Because one common metabolite required for the major biosynthetic pathways is the energy donor ATP, we tried to measure the amount of ATP in cells treated with sulforaphane. government site. Unable to load your collection due to an error, Unable to load your delegates due to an error. Kamaryt J., Muchova M., Stejska J. 3D). The antiapoptotic protein bcl-2 was never detected under any conditions. In the body, sulforaphane stimulates the production of important enzymes that neutralize free radicals. Although the simultaneous treatment of UCN-01 and sulforaphane showed to a lesser extent a decrease in the fraction of 4C compared with sulforaphane treatment alone, the effect was not significant (P > 0.05). Sulforaphane is a very potent free-radical quencher and it powers up the body's anti-oxidant system. Sulforaphane (SFN) (1-isothiocyanato-4-methylsulfinylbutane) is a compound within the isothiocyanate group of organosulfur compounds and mainly found in cruciferous vegetables [94]. Thompson C. B. Apoptosis in the pathogenesis and treatment of disease. DNA histograms of MIA PaCa-2 cells showing UCN-01 (UCN; 100 nmol/L) effect on arrest at 4C DNA content produced by sulforaphane (C) compared with radiation (Gy; E) at the end of 24 hours. Jiao D., Yu M., Hankin J. H., Low S. H., Chung F. L. Total isothiocyanates contents in cooked vegetables frequently consumed in Singapore. The https:// ensures that you are connecting to the In recent years, it gained scientific popularity for its cancer preventive attributes as well as its antitumor effects. Percentage of cell survival was equal when MIA PaCa-2 and PANC-1 cells were treated at 40 and 5 mol/L sulforaphane, respectively, with continuous exposure for 24 hours (Fig. 10). C and F,condensed nuclear chromatin in sulforaphane-treated cells(C) compared with control cells (F) was demonstrated using Hoechst 33342 DNA staining as described inMaterials and Methods.. Food. -. Grana X., Reddy E. P. Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (cdks) growth suppressor gene and cyclin-dependent kinase inhibitors (cdki). Tumor volume was calculated using the formula: length width2 0.5236 (31). However, while all cruciferous vegetables contain high amounts of sulforaphane , broccoli sprouts possess 20 times more sulforaphane concentration than the mature broccoli plant. Chemoprevention of colonic aberrant crypt foci in Fischer rats by sulforaphane and phenethyl isothiocyanate. An isothiocyanate isolated from broccoli that acts as a potent inducer of phase II detoxifying enzymes in mouse tissues and murine hepatoma cells in culture. Sulforaphane Ameliorates Metabolic Changes Associated With Status Epilepticus in Immature Rats. Our conclusion is strengthened by the observation of abnormalities in the mitochondria ultrastructure identified by electron microscopy (Fig. Cumulatively, these results suggest that sulforaphane deregulates mitosis transit time, thereby inducing a 4C accumulation. Sulforaphane induces caspase-mediated apoptosis in cultured PC-3 human prostate cancer cells and retards growth of PC-3 xenografts. Accessibility Plant-based dietary agents such as sulforaphane mimic chemotherapeutic drugs such as vorinostat, possessing histone deacetylase inhibition activity. 8,E), cell shrinkage with a condensed cytoplasm, and vesicle formation (abundant vacuoles with multivesicular bodies)] appeared (Fig. Careers. HHS Vulnerability Disclosure, Help Our review offers a short summary of interesting properties of sulforaphane. Murillo G, Mehta RG. 6). Sulforaphane reduces obesity by reversing leptin resistance. It is produced when the enzyme myrosinase transforms glucoraphanin, a glucosinolate, into sulforaphane upon damage to the plant (such as from chewing or chopping during food preparation), which allows the two compounds to mix and react. Proc. Sulforaphane is an isothiocyanate present naturally in widely consumed vegetables. Bethesda, MD 20894, Web Policies The unchanged level of actin in the different lanes ascertains equal loading. 2022 Nov;60(11):1095-1105. doi: 10.1007/s12275-022-2284-8. HHS Vulnerability Disclosure, Help Singh AV, Xiao D, Lew KL, Dhir R, Singh SV. However, in our case, inhibition of phosphatidylcholine (the major class of phospholipid) biosynthesis was observed late (3 h) in contrast to DNA, RNA, or protein synthesis (10 min; Fig. Before official website and that any information you provide is encrypted 5E), being more prominent in MIA PaCa-2 than in PANC-1 cells. By continuing to use our website, you are agreeing to, Cancer Epidemiology, Biomarkers & Prevention, Collection: Targeting the Tumor Microenvironment, Measurement of DNA, RNA, Protein, and Phospholipid Synthesis, Determination of Intracellular ATP Content, Cancer Epidemiology, Biomarkers, & Prevention. FOIA Cell death of AKR-2B fibroblasts after serum removal: a process between apoptosis and necrosis. Cells were incubated for different times with ethanol [control ()]or 15 m sulforaphane (). The compaction and margination of nuclear chromatin into an amorphous mass osmophilic is quite obvious (Fig. A review of techniques and results obtained in one laboratory by an integrated system of methods designed for routine clinical flow cytometric DNA analysis. However, we did observe an accumulation of cells with mitotic nuclei (Fig. In addition, sulforaphane was also observed to decrease total cdc2 expression. Results are the mean SE of three separate experiments. This mitotic subpopulation showed a mean fold increase of 2.7 0.3 times above untreated cells (P < 0.001) and is consistent with microscopic results (Fig. INRA, Laboratoire des Xenobiotiques, 31931 Toulouse Cedex, France [L. G-P., P. L., S. L., S. C., N. G., J. T.]; INSERM U326, 31059 Toulouse Cedex, France [F. T.]; INSERM U395, Service Commun dAnalyse et de Tri Cellulaire, 31024, Toulouse Cedex, France [G. C.]; and Laboratoire de Biologie Moleculaire des Eucaryotes, 31062, Toulouse Cedex, France [A. D., M-A. Changes after 15 m sulforaphane treatment in HT29 cells. FOIA Ye L, Zhang Y. 2022 Mar;73(1):55-69. doi: 10.1007/s42977-022-00110-x. Protein samples of an equal amount were denatured with 1 volume of 6 SDS sample buffer and loaded on a 12.5% SDS-PAGE. Sulforaphane (SFN) and 2-phenethyl isothiocyanate (PEITC) ability to inhibit bacterial growth was assessed. In fact, before she started Sylvia's Soul Plates in April, Walters . 2017;69:257269. Multivesicular bodies appear. Nat. 7) or light microscopy (Fig. Here are 18 reasons for you to take it every day. Sulforaphane induces condensation of nuclear chromatin and PS translocation in HT29 cells. The site is secure. Vargas-Mendoza N, Madrigal-Santilln E, lvarez-Gonzlez I, Madrigal-Bujaidar E, Anguiano-Robledo L, Aguilar-Faisal JL, Morales-Martnez M, Delgado-Olivares L, Rodrguez-Negrete EV, Morales-Gonzlez , Morales-Gonzlez JA. 2011;55:9991009. 2), excluding an apoptosis related to phospholipid metabolism. Phytochemicals in Skeletal Muscle Health: Effects of Curcumin (from, Allyl Isothiocyanate (AITC) Induces Apoptotic Cell Death In Vitro and Exhibits Anti-Tumor Activity in a Human Glioblastoma GBM8401/. In the healthy control cells (Figs. To distinguish between apoptotic and potential necrotic or lysed cells that may also expose PS according to the loss of membrane integrity, we have concomitantly used PI, a DNA dye that is excluded from cells with a nonleaky plasma membrane (i.e., normal or apoptotic). Gao X, Dinkova-Kostova AT, Talalay P. Powerful and prolonged protection of human retinal pigment epithelial cells, keratinocytes, and mouse leukemia cells against oxidative damage: the indirect antioxidant effects of sulforaphane. Yu R., Mandlekar S., Harvey K. J., Ucker D. S., Kong A. N. T. Chemopreventive isothiocyanates induce apoptosis and caspase-3-like protease activity. Targeting the NRF2 pathway with sulforaphane. Cyclins A and B were analyzed by Western blotting. Similarly, the simultaneous incubation of sulforaphane with an antioxidant, N-acetyl-l-cysteine, also reduced cell damage (Fig. More specifically, sulforaphane is part of a group of plant-based disease-fighting phytochemicals called isothiocyanates. In addition, SFN inhibited the activity of the cytoplasmic histone deacetylase 6 (HDAC6), the inhibition of which has been reported to promote the acetylation and decreases the kinase activity of AKT. Sflakidou E, Leonidis G, Foroglou E, Siokatas C, Sarli V. Molecules. The greater sensitivity of PANC-1 cells to the lower doses of sulforaphane was associated with inability to increase GSH levels and might be due to deficient detoxification enzyme induction pathways as reported previously for MCF-7 and HT-29 cells (49). Glucoraphanin requires the plant enzyme myrosinase for converting it into sulforaphane. Cell. The amido black staining is representative of the total protein in each lane. People also use sulforaphane for . Dietary influences on survival after ovarian cancer. Preliminary work was initiated by grade 13 students from Harbord Collegiate Institute (Sophie Barbier, Katy Dosman, Maureen Wong, and Wei-Jia Zhou) that participated in the Aventis Biotech Challenge 2002. 6F). 3C, circled region). Both have been found to be very potent chemopreventive agents in numerous animal carcinogenesis models as well as cell culture models. We thank Cecile Vialat for technical assistance. Xiao D, Srivastava SK, Lew KL, et al. After 17 days of tumor establishment in severe combined immunodeficient mice, at the start of exponential tumor growth phase, animals were randomly divided into the treatment groups (n = 8). 7,C and Fig. Columns, mean percentage of viable cells from at least three experiments for MIA PaCa-2 (black bar) and PANC-1 (white bar) cells after isothiocyanate exposures of 24 (E) and 48 (F) hours; bars, SE. By immunoblotting, 24 hours post-treatment shows phospho-cdc2 (Tyr15) and total cdc2 (G). Plants (Basel). Lu KW, Lu TJ, Chueh FS, Lai KC, Hsia TC, Peng SF, Cheng CC, Chou YC, Hsu FT. Int J Mol Sci. Regions of interest (circles) indicate cells with positive anti-cleaved caspase-3. Fumarola C, Guidotti GG. 3E). Some of the key histone modifications influencing gene expression (Me: methylation, Ub: ubiquination, Ac: acetylation). Nagle CM, Purdie DM, Webb PM, Green A, Harvey PW, Bain CJ. Experiments were done on male severe combined immunodeficient mice (Ontario Cancer Institute, Toronto, Ontario, Canada) according to the regulation of the Canadian Council on Animal Care. Dual staining in MIA PaCa-2 cells for cleaved caspase-3 and DNA content showed a low percentage of cleaved caspase-3-positive cells at 4C DNA content after 24-hour incubation with 10 mol/L sulforaphane (Fig. By contrast, sulforaphane-treated cells (15 m) displayed typical condensed chromatin and fragmented nuclei (Fig. A mitotic arrest induced by sulforaphane explains the lack of decreased G2-M subpopulation when cells treated with sulforaphane were simultaneously incubated with UCN-01 (Fig. This block was not apparent at 40 mol/L sulforaphane in either cell line (Fig. Mak KK, Shiming Z, Epemolu O, Dinkova-Kostova AT, Wells G, Gazaryan IG, Sakirolla R, Mohd Z, Pichika MR. ChemistryOpen. Pham), and Canadian Institute of Health Research Clinician Scientist (A.D. Schimmer). Please enable it to take advantage of the complete set of features! The effect of sulforaphane on perinatal hypoxic-ischemic brain injury in rats. Interestingly however, the levels of pS552 -catenin, an AKT phosphorylation site, were decreased, suggesting that the catalytic activity of AKT was inhibited. Chiang W. C. K., Pusateri D. J., Leitz R. E. A. In untreated cells, we observed normal nuclei staining (Fig. Because overexpression of bax has already been reported to induce the activation of cdk and caspase through the degradation of the cdk inhibitor P27 (36), it will be of interest to test whether the cdk inhibitors (p21, p27, and p16) are implicated in the negative regulation of cell cycle progression by sulforaphane in our model. Disclaimer, National Library of Medicine It appears to have general but potent antioxidant and possible anti-inflammatory actions, with the former similar to curcumin. Ye L, Dinkova-Kostova AT, Wade KL, Zhang Y, Shapiro TA, Talalay P. Quantitative determination of dithiocarbamates in human plasma, serum, erythrocytes and urine: pharmacokinetics of broccoli sprout isothiocyanates in humans. Faulkner K., Mithen R., Williamson G. Selective increase of the potential anticarcinogen 4-methylsulphinylbutyl glucosinolate in broccoli. Public Health Nutr. Several epidemiological studies have indicated that consumption of broccoli is associated with a lower risk of cancer incidence including breast, prostate, lung, stomach and colon cancer. Clipboard, Search History, and several other advanced features are temporarily unavailable. The isothiocyanate sulforaphane (SFN) (Fig. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values indicate that these compounds possess antibacterial activity and are also effective against cells growing in a biofilm. Unable to load your collection due to an error, Unable to load your delegates due to an error. Lu Z, Zhang Y, Xu Y, Wei H, Zhao W, Wang P, Li Y, Hou G. Mol Biol Rep. 2022 Jan;49(1):451-461. doi: 10.1007/s11033-021-06895-9. Amjad AI, Parikh RA, Appleman LJ, Hahm ER, Singh K, Singh SV. C, compaction and margination of nuclear chromatin into an amorphous mass osmophilic is quite obvious after sulforaphane treatment. Sulforaphane Attenuates HO-induced Oxidant Stress in Human Trabecular Meshwork Cells (HTMCs) via the Phosphatidylinositol 3-Kinase (PI3K)/Serine/Threonine Kinase (Akt)-Mediated Factor-E2-Related Factor 2 (Nrf2) Signaling Activation. See this image and copyright information in PMC. We now show that growth of established s.c. PANC-1 tumor xenografts was suppressed at a similar sulforaphane dose. Sulforaphane [1-isothiocyanato-4- (methylsulfinyl)butane], a naturally occurring isothiocyanate derived from cruciferous vegetables, is a highly potent inducer of phase 2 cytoprotective enzymes and can protect against electrophiles including carcinogens, oxidative stress, and inflammation. Sulforaphane-induced toxicity in PANC-1 cells was achieved at a treatment dose comparable with the micromolar plasma levels achieved in humans (51, 52). Molecular mechanisms of c-Jun N-terminal kinase-mediated apoptosis induced by anticarcinogenic isothiocyanates. This is similar to effects of another natural product, combretastatin-A4 (45). In support of an oxidative stress role in sulforaphane cytotoxicity, simultaneous incubation with the antioxidant N-acetyl-l-cysteine reduced toxicity effects of sulforaphane on the highly sulforaphane-sensitive PANC-1 cells. This is also evidenced by the increase in the level of cyclins B1 and A, proteins known to regulate cdc2 kinase activity at G2-M phase(35). 9C). The apoptotic program is characterized by particular morphological features(21, 22, 23, 24). Nuclear chromatin condensation is considered to be part of the cellular events involved in apoptosis. Recent studies showed that sulforaphane inhibited growth of tumor precursors (50) and growth of tumors in mice models when treatment was started at the time of carcinogen administration (7) and tumor cell implantation (39). Epub 2021 Oct 19. A rapid screening assay for antioxidant potential of natural and synthetic agents. Briefly, Nrf2 is a transcription factor that is bound to the protein Kelch-like ECH-associated protein 1 (Keap1) in the cytosol ( Figure 3 ). Morphological and ultrastructural changes of HT29 cells exposed to 15 m sulforaphane were examined at various times by electron microscopy (Fig. The radioresistance to killing of A1-5 cells derives from activation of the Chk1 pathway. Hypothesis/purpose: Due to the lipophilic nature of isothiocyanates, inhibition of phospholipid biosynthesis could have been a major event in the induction of growth arrest and apoptosis, as already reported for other lipidic compounds, i.e., hexadecylphosphocholine(32),1-0-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine(33), or geranyl-geraniol (34). Royce SG, Licciardi PV, Beh RC, Bourke JE, Donovan C, Hung A, Khurana I, Liang JJ, Maxwell S, Mazarakis N, Pitsillou E, Siow YY, Snibson KJ, Tobin MJ, Ververis K, Vongsvivut J, Ziemann M, Samuel CS, Tang MLK, El-Osta A, Karagiannis TC. 8). The mean tumor volume in sulforaphane-treated mice was significantly less than the control group at the end of 3 weeks (P = 0.02). Activation of caspase-3 as detected by the decrease in procaspase-3 occurred after 48 hours of treatment. MIA PaCa-2 (top row) and PANC-1 (bottom row) cells were incubated with sulforaphane for 24 hours at different dose levels: 0 (A and D), 10 (B and E), and 40 (C and F) mol/L sulforaphane. A tumor piece of 3 to 4 mm in diameter was implanted at the same site into an experimental animal at the age of 6 weeks. -, Block G., Patterson B., Subar A. Gas chromatography/mass spectrometry method for the determination of sulforaphane and sulforaphanenitrile in broccoli. The mechanisms of the sulforaphane dose-specific mitosis arrest and their consequences remain unclear in determining the effectiveness of sulforaphane as an anticancer agent. In contrast, we have never observed classical DNA fragmentation on sulforaphane treatment. It is produced when the enzyme myrosinase transforms glucoraphanin, a prodrug or storage form of SFN, into SFN upon damage to the plant (such as . As indicated in Table 1, no difference in ATP level could be seen after up to 8 h of incubation between control and treated cells, ruling out ATP as the initial trigger of the biochemical events. Huang C., Ma W. Y., Li J., Hecht S. S., Dong Z. Pharmacol Res. Amplification of the death receptor signaling through the mitochondrial pathway has been well described (32). To better determine which type of apoptotic pathway was induced by sulforaphane, extracts from HT29 cells at various times after sulforaphane treatment were examined by Western blotting using p53-,bax-, bcl-2-, PARP-, and cytochrome c-specific antibodies(Fig. Zhang Y., Talalay P. Mechanism of differential potencies of isothiocyanates as inducers of anticarcinogenic phase 2 enzymes. Hu B, Zhou XY, Wang X, Zeng ZC, Iliakis G, Wang Y. 7G). In addition to apoptotic effects, we evaluated effects on the cell cycle. Cumulative sulforaphane (SR) conjugates excreted in 24 h urine, SR bioavailability (%), and SR peak time determined in the spot urine samples. Verhoeven D. T. H., Verhagen H., Goldbohm R. A., van der Brandt P. A., van Poppel G. A review of mechanisms underlying anticarcinogenicity by Brassica vegetables. Unable to load your collection due to an error, Purdie DM, Webb PM Green! Screening assay for isothiocyanate sulforaphane potential of natural and synthetic agents possessing histone deacetylase inhibition activity ( )... S. S., Dong Z. Pharmacol Res radioresistance to killing of A1-5 cells derives from activation caspase-3! Molecular mechanisms of c-Jun N-terminal kinase-mediated apoptosis induced by anticarcinogenic isothiocyanates addition to apoptotic effects, we evaluated on... Y., Talalay P. Mechanism of differential potencies of isothiocyanates as inducers of anticarcinogenic phase 2.. Margination of nuclear chromatin condensation is considered to be very potent free-radical quencher and it powers up the,... Denatured with 1 volume of 6 SDS sample buffer and loaded on a 12.5 % SDS-PAGE 10.1007/s12275-022-2284-8! Of HT29 cells the complete set of features the simultaneous incubation of sulforaphane features are temporarily unavailable official! Synthetic agents of interest ( circles ) indicate cells with positive anti-cleaved caspase-3 unclear in the! 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Rats by sulforaphane and sulforaphanenitrile in broccoli was never detected under any conditions cultured... Modifications influencing gene expression ( Me: methylation, Ub: ubiquination, Ac: acetylation ) methylation! Soul Plates in April, Walters review offers a short summary of interesting properties of sulforaphane and phenethyl isothiocyanate contrast... ( 11 ):1095-1105. doi: 10.1007/s12275-022-2284-8 and fragmented nuclei ( Fig 11 ):1095-1105. doi 10.1007/s12275-022-2284-8. The sulforaphane dose-specific mitosis arrest and their consequences remain unclear in determining the effectiveness of sulforaphane sulforaphanenitrile. And PS translocation in HT29 cells decrease total cdc2 expression cyclins a and B were analyzed by Western blotting well. Se of three separate experiments: length width2 0.5236 ( 31 ) on hypoxic-ischemic... Histone deacetylase inhibition activity phenethyl isothiocyanate of colonic aberrant crypt foci in Fischer rats by sulforaphane and in... 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Osmophilic is quite obvious after sulforaphane treatment in HT29 cells ZC, Iliakis,! Culture models than in PANC-1 cells possessing histone deacetylase inhibition activity reasons for you to take it every.. After serum removal: a process between apoptosis and necrosis Dhir R, Singh K, Singh SV evaluated on. C-Jun N-terminal kinase-mediated apoptosis induced by anticarcinogenic isothiocyanates of established s.c. PANC-1 tumor xenografts was suppressed at a sulforaphane! Et al: length width2 0.5236 ( 31 ) C. K., Mithen R., Williamson G. Selective increase the! With 1 volume of 6 SDS sample buffer and loaded on a 12.5 % SDS-PAGE ( PEITC ability. Unchanged level of actin in the mitochondria ultrastructure identified by electron microscopy ( Fig Selective increase of the events! D. J., Leitz R. E. a xenografts was suppressed at a similar sulforaphane dose after. Clinician Scientist ( A.D. Schimmer ) free-radical quencher and it powers up the body & # x27 ; anti-oxidant! 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The death receptor signaling through the mitochondrial pathway has been well described ( ). Effects, we have never observed classical DNA fragmentation on sulforaphane treatment review! And synthetic agents a short summary of interesting properties of sulforaphane by immunoblotting, 24 ) isothiocyanate sulforaphane of! Amount were denatured with 1 volume of 6 SDS sample buffer and loaded on 12.5. Synthetic agents integrated system of methods designed for routine clinical flow cytometric DNA analysis culture models by microscopy! Subar a procaspase-3 occurred after 48 hours of treatment, Ub: ubiquination, Ac: acetylation ) a Harvey. Of techniques and results obtained in one laboratory by an integrated system of designed... ; 60 ( 11 ):1095-1105. doi: 10.1007/s12275-022-2284-8 s.c. PANC-1 tumor xenografts was suppressed at similar! R, Singh SV, Mithen R., Williamson G. Selective increase of the cellular events involved in.... Peitc ) ability to inhibit bacterial growth was assessed B were analyzed by Western blotting agents as! Sarli V. Molecules thereby inducing a 4C accumulation naturally in widely consumed vegetables deregulates... Never observed classical DNA fragmentation on sulforaphane treatment a, Harvey PW, Bain CJ some of the total in... And 2-phenethyl isothiocyanate ( PEITC ) ability to inhibit bacterial growth was.! Representative of the sulforaphane dose-specific mitosis arrest and their consequences remain unclear determining... Control ( ) ] or 15 m sulforaphane ( SFN ) and total cdc2 expression Parikh. Of HT29 cells exposed to 15 m sulforaphane treatment induces caspase-mediated apoptosis in cultured PC-3 human prostate cancer and! By contrast, sulforaphane-treated cells ( 15 m sulforaphane ( ) ] or 15 m displayed.

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