Brassinosteroids Abstracts 1

© 2011

24-Epibrassinolide, a Phytosterol from the Brassinosteroid Family, Protects Dopaminergic Cells

         (Carange, Longpre ́ et al. 2011) Download

Oxidative stress and apoptosis are frequently cited to explain neuronal cell damage in various neurodegenerative disorders, such as Parkinson’ s disease. Brassinosteroids (BRs) are phytosterols recognized to promote stress tolerance of vegetables via modulation of the antioxidative enzyme cascade. However, their antioxidative effects on mammalian neuronal cells have never been examined so far. We analyzed the ability of 24-epibrassinolide (24-Epi), a natural BR, to protect neuronal PC12 cells from 1-methyl-4-phenylpyridinium- (MPP+-) induced oxidative stress and consequent apoptosis in dopaminergic neurons. Our results demonstrate that 24-Epi reduces the levels of intracellular reactive oxygen species and modulates superoxide dismutase, catalase, and glutathione peroxidase activities. Finally, we determined that the antioxidative properties of 24-Epi lead to the inhibition of MPP+-induced apoptosis by reducing DNA fragmentation as well as the Bax/Bcl-2 protein ratio and cleaved caspase-3. This is the first time that the potent antioxidant and neuroprotective role of 24-Epi has been shown in a mammalian neuronal cell line.

3-Keto-22-epi-28-nor-cathasterone, a brassinosteroid-related metabolite from Cystoseira myrica

            (Hamdy, Aboutabl et al. 2009) Download

Bioassay-guided purification of an ethanolic extract of Cystoseira myrica against HEPG-2 (liver) and HCT116 (colon) human cancer cell lines led to the isolation of 3-keto-22-epi-28-nor-cathasterone, 1 and cholest-4-ene-3,6-dione, 2. This finding allowed us to report for the first time that a brassinosteroid-related metabolite occurs in seaweed. These compounds showed activity in the range of 12.38-1.16 microM with selective activity of compound 2 to liver cancer cell lines.


Anticancer and antiproliferative activity of natural brassinosteroids

            (Malikova, Swaczynova et al. 2008) Download

Brassinosteroids (BRs) are steroid plant hormones that are essential for many plant growth and developmental processes, including cell expansion, vascular differentiation and stress responses. Up to now the inhibitory effects of BRs on cell division of mammalian cells are unknown. To determine basic anticancer structure-activity relationships of natural BRs on human cells, several normal and cancer cell lines have been used. Several of the tested BRs were found to have high cytotoxic activity. Therefore, in our next series of experiments, we tested the effects of the most promising and readily available BR analogues with interesting anticancer properties, 28-homocastasterone (1) and 24-epibrassinolide (2), on the viability, proliferation, and cycling of hormone-sensitive/insensitive (MCF-7/MDA-MB-468) breast and (LNCaP/DU-145) prostate cancer cell lines to determine whether the discovered cytotoxic activity of BRs could be, at least partially, related to brassinosteroid-nuclear receptor interactions. Both BRs inhibited cell growth in a dose-dependent manner in the cancer cell lines. Flow cytometry analysis showed that BR treatment arrested MCF-7, MDA-MB-468 and LNCaP cells in G(1) phase of the cell cycle and induced apoptosis in MDA-MB-468, LNCaP, and slightly in the DU-145 cells. Our results provide the first evidence that natural BRs can inhibit the growth, at micromolar concentrations, of several human cancer cell lines without affecting the growth of normal cells. Therefore, these plant hormones are promising leads for potential anticancer drugs.

Brassinosteroids: synthesis and activity of some fluoro analogues

            (Slavikova, Kohout et al. 2008) Download

Three types of 5alpha-androstane and ergostane analogues of brassinolide, containing a fluorine atom in either the 3alpha or the 5alpha positions or in 3alpha and 5alpha positions, were prepared using standard operations (reaction of 3beta-alcohols with (diethylamino)sulfur trifluoride, cleavage of epoxide with HF in py or BF 3.Et 2O). The 5alpha-fluorine was found to affect chemical reactivity (e.g., electrophilic addition to the Delta (2)-double bond) as well as physical properties (e.g., NMR, chromatographic behavior) of the products. Cytotoxicity of the products was studied using human normal and cancer cell lines with 28-homocastasterone as positive control and their brassinolide type activity was established using the bean second-internode test with 24-epibrassinolide as standard. The equivalence of F and OH groups was observed in some of the active compounds. The anticancer and the brassinolide-type activity do not correlate with each other: ergostane derivatives were most active in the former test while androstane derivatives were best in the latter.

Brassinosteroids cause cell cycle arrest and apoptosis of human breast cancer cells

            (Steigerova, Oklestkova et al. 2011) Download

Brassinosteroids (BRs) are plant hormones that appear to be ubiquitous in both lower and higher plants. Recently, we published the first evidence that some natural BRs induce cell growth inhibitory responses in several human cancer cell lines without affecting normal non-tumor cell growth (BJ fibroblasts). The aim of the study presented here was to examine the mechanism of the antiproliferative activity of the natural BRs 28-homocastasterone (28-homoCS) and 24-epibrassinolide (24-epiBL) in human hormone-sensitive and -insensitive (MCF-7 and MDA-MB-468, respectively) breast cancer cell lines. The effects of 6, 12 and 24h treatments with 28-homoCS and 24-epiBL on cancer cells were surveyed using flow cytometry, Western blotting, TUNEL assays and immunofluorescence analyses. The studied BRs inhibited cell growth and induced blocks in the G(1) cell cycle phase. ER-alpha immunoreactivity was uniformly present in the nuclei of control MCF-7 cells, while cytoplasmic speckles of ER-alpha immunofluorescence appeared in BR-treated cells (IC(50), 24h). ER-beta was relocated to the nuclei following 28-homoCS treatment and found predominantly at the periphery of the nuclei in 24-epiBL-treated cells after 24h of treatment. These changes were also accompanied by down-regulation of the ERs following BR treatment. In addition, BR application to breast cancer cells resulted in G(1) phase arrest. Furthermore, TUNEL staining and double staining with propidium iodide and acridine orange demonstrated the BR-mediated induction of apoptosis in both cell lines, although changes in the expression of apoptosis-related proteins were modulated differently by the BRs in each cell line. The studied BRs seem to exert potent growth inhibitory effects via interactions with the cell cycle machinery, and they could be highly valuable leads for agents for managing breast cancer.

Brassinolide, a plant sterol from pollen of Brassica napus L., induces apoptosis in human prostate cancer PC-3 cells

            (Wu and Lou 2007) Download

Brassinolide is a plant sterol first isolated from pollen of rape (Brassica napus L.). The present study was carried out to investigate the effect of brassinolide on androgen-independent human prostate cancer PC-3 cell viability. Results showed that brassinolide could induce a time and concentration-dependent cytotoxicity in PC-3 cells. The mode of cell death appeared to be predominately apoptosis, as shown by flow-cytometric analysis, fluorescence and transmission electron microscopes. Caspase-3 activity was obviously increased after brassinolide treatment. Western blot studies indicated that treatment with brassinolide triggered a time-dependent decrease in the expression of anti-apoptotic protein Bcl-2. We suggest that brassinolide could induce cytotoxicity in PC-3 cells by triggering apoptosis. Brassinolide might therefore be a promising candidate for the treatment of prostate cancer.


References

Carange, J., F. Longpre ́, et al. (2011). "24-Epibrassinolide, a Phytosterol from the Brassinosteroid Family, Protects Dopaminergic Cells." J Toxicology.

Hamdy, A. H., E. A. Aboutabl, et al. (2009). "3-Keto-22-epi-28-nor-cathasterone, a brassinosteroid-related metabolite from Cystoseira myrica." Steroids 74(12): 927-30.

Malikova, J., J. Swaczynova, et al. (2008). "Anticancer and antiproliferative activity of natural brassinosteroids." Phytochemistry 69(2): 418-26.

Slavikova, B., L. Kohout, et al. (2008). "Brassinosteroids: synthesis and activity of some fluoro analogues." J Med Chem 51(13): 3979-84.

Steigerova, J., J. Oklestkova, et al. (2011). "Brassinosteroids cause cell cycle arrest and apoptosis of human breast cancer cells." Chem Biol Interact 188(3): 487-96.

Wu, Y. D. and Y. J. Lou (2007). "Brassinolide, a plant sterol from pollen of Brassica napus L., induces apoptosis in human prostate cancer PC-3 cells." Pharmazie 62(5): 392-5.