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Mechanisms of Anticancer Activity of Sulforaphane from Brassica oleracea in HEp-2 Human Epithelial Carcinoma Cell Line

  • Devi, J. Renuka (Department of Biotechnology, School of Bioengineering, SRM University) ;
  • Thangam, E. Berla (Department of Biotechnology, School of Bioengineering, SRM University)
  • Published : 2012.05.30

Abstract

Sulforaphane (SFN) an isothiocyanate formed by hydrolysis of glucosinolates found in Brassica oleraceae is reported to possess anticancer and antioxidant activities. In this study, we isolated SFN from red cabbage (Brassica oleraceae var rubra) and evaluated the comparative antiproliferative activity of various fractions (standard SFN, extract and purified SFN) by MTT assay in human epithelial carcinoma HEp -2 and and Vero cells. Probable apoptotic mechanisms mediated through p53, bax and bcl-2 were also examined. The SFN fraction was collected by HPLC, enriched for its SFN content and confirmed. Expression of apoptosis-related proteins was detected by western blotting and RT PCR. Results showed that Std SFN and purified SFN concentration found to have closer $IC_{50}$ which is equal to 58.96 microgram/ml (HEp-2 cells), 61.2 microgram/ml (Vero cells) and less than the extract which is found to be 113 microgram/ml (HEp-2 cells) and 125 microgram/ml (Vero cells). Further studies on apoptotic mechanisms showed that purified SFN down-regulated the expression of bcl-2 (antiapoptotic), while up-regulating p53 and Bax (proapoptotic) proteins, as well as caspase-3. This study indicates that purified SFN possesses antiproliferative effects the same as Std SFN and its apoptotic mechanism in HEp-2 cells could be mediated through p53 induction, bax and bcl-2 signaling pathways.

Keywords

References

  1. Aamer Qazi, Pal J, Maitah M, et al (2010). Anticancer activity of a Broccoli derivative sulforaphane in Barrett adenocarcinoma potential use in cheoprevention and as adjuvant in Chemotherapy. Translational Oncol, 3, 389-99. https://doi.org/10.1593/tlo.10235
  2. Bonnesen C, Eggleston I, Hayes JD (2001). Dietary indoles and Isothiocyanates that are generated from Cruciferous vegetables can both stimulate apoptosis and confer protection against DNA damage in human colon cell lines. Cancer Res, 61, 6120-30.
  3. Brandi G, Schiavano GF, Marco CD, et al (2005). Mechanisms of action and antiproliferative properties of Brassica oleracea juice in human Breast cancer cell lines. J Nutri, 135, 1504-9.
  4. Cawthon RM ( 2002). Telomere measurement by Quantitative PCR. Nucleic Acids Res, 30, e47. https://doi.org/10.1093/nar/30.10.e47
  5. Dashwood RH (1988). Indole-3carbinol: Anticarcinogen or tumor promoter in brassica vegetables? Chem Biol Inter, 110, 1-5.
  6. Davis CD, Zeng H, Finley JE (2002). Selenium Enriched Broccoli decreases intestinal Tumorigenesis in Multiple intestinal Neoplasia. J Nutr, 132, 302-9.
  7. Denoyelle C, Aibanese P, Li G, et al (2003). Molecular mechanism of the anticancer activity of cerivastatin an inhibitor of HMG Co A reductase on aggressive human breast cancer cells. Cell Signal, 15, 327-38. https://doi.org/10.1016/S0898-6568(02)00124-9
  8. Edmondson JM, Armstrong LS, Martinez AO (1988). A rapid and simple MTT based spectrophotometric assay for determining drug sensitivity in monolayer cultures. J Tissue Culture Methods, 2, 15-7.
  9. Fimognari C, Niisse M, Cesari R, et al (2002). Growth inhibition cell cycle arrest and apoptosis in human T cell leukemia by the isothiocynate Sulforaphane. Carcinogenesis, 23, 581-6. https://doi.org/10.1093/carcin/23.4.581
  10. Firestone GL, Bjeldanes LF (2003). Indole 3 carbinol and 3-3' indolyl methane signalling pathways control cell cycle Gene transcription in Human Breast Cancer cells by regulating promoter-Sp1 Transcription factor interactions. J Nutri, 133, 2448-55
  11. Gamet-Payrastre, Li, Solange Lumeau (2000). Sulforaphane,a Naturally Occurring lsothiocyanate, induces Cell Cycle Arrest and Apoptosis in HT29 Human Colon Cancer Cells. Cancer Research, 60, 1426-33.
  12. Gasper AV, Traka M, Bacon JR, et al (2007). Consuming broccoli does not induce genes associated with xenobiotic metabolism and cell cycle control in human gastric mucosa. J Nutr, 137, 1718-24. https://doi.org/10.1093/jn/137.7.1718
  13. Han JM, Lee YJ, Lee SY, et al (2007). Protective effect of sulforaphane against dopaminergic cell death. J Pharmacol Exp Ther, 321, 249-56. https://doi.org/10.1124/jpet.106.110866
  14. Jin CY, Moon DO, Moon JDL, et al (2007). Sulforaphane sensitizes tumor necrosis factor related apoptosis induced ligand mediated apoptosis through downregulation of ERK and AKT in Lung adenocarcinoma A549 cells. Carcinogenesis, 28, 1058-66.
  15. Liang H, Yuan QP, Dong HR, Liu YM (2006). Determination of sulforaphane in broccoli and cabbage by high-performance liquid chromatography. J Food Composition Analysis, 19, 473-6. https://doi.org/10.1016/j.jfca.2005.11.005
  16. lllic N, Normanly J, Cohen JD (1996). Quantification of free plus conjugated indoleacetic acid in arabidopsis requires correction for the nonenzymatic conversion of indolic nitriles. Plant Physiol, 11, 781-8.
  17. Matsui TA, Sowa Y, Yoshida T, et al (2006). Sulforaphane enhances TRAIL induced apoptosis through the induction of DR5 expression in human Osteosarcoma cells. Carcinogenesis, 27, 1768-77. https://doi.org/10.1093/carcin/bgl015
  18. McNaughton SA, Marks GC (2003). Development of a food composition database for the estimation of dietary intakes of glucosinolates, the biologically active constituents of cruciferous vegetables. Bri J Nutr, 90, 687-97. https://doi.org/10.1079/BJN2003917
  19. Meyer M, Adam ST (2008). Comparison of glucosinolate levels in commercial broccoli and red cabbage from conventional and ecological farming. Eur Food Res Technol, 226, 1429-37. https://doi.org/10.1007/s00217-007-0674-0
  20. Chen MJ, Tang WY, Hsu CW, et al (2012). Apoptosis induction in primary human colorectal cancer cell lines and retarded tumor growth in SCID mice by sulforaphane. Evid Based Complement Alternat Med, 2012, 415231,
  21. Myzak MC, Hardin K, Wang R, Dashwood RH, Ho E (2006). Sulforaphane inhibits histone deacetylase activity in BPH-1, LnCaP and PC-3 prostate epithelial cells. Carcinogenesis, 27, 811-9. https://doi.org/10.1093/carcin/bgi265
  22. Pappa G, Lichtenberg M, Lori R, et al (2006). Comparision of growth inhibition profiles and mechanisms of apoptosis induction in human colon cancer cell lines by isothiocyanates and indoles from Brassicaceae. Mutation Res, 599, 76-87. https://doi.org/10.1016/j.mrfmmm.2006.01.007
  23. Pledgie-Tracy A, Sobolewski MD, Davidson NE (2007). Sulforaphane induces cell type specific apoptosis in human breast cancer cell lines. Molecular cancer therapeutics, 6, 1013-21. https://doi.org/10.1158/1535-7163.MCT-06-0494
  24. Renuka Devi J, Berla Thangam E (2010 ). Extraction and Separation of Glucosinolates from Brassica Oleraceae Rubra. Adv Biol Res, 4, 309-13.
  25. Riby JE, Xue L, Cahtterji U, et al (2006). Activation and potentiation of interferon-gamma signaling by 3,3'-diindolylmethane in MCF-7 breast cancer cells. Mol Pharmacol, 69, 430-9.
  26. Savli H, Sirma S, Nagy B, et al (2004).Real-Time PCR analysis of af4 and dek genes expression in acute promyelocytic leukemia t (15;17) patients. Exp Mol Med, 36, 279-82. https://doi.org/10.1038/emm.2004.38
  27. Sreenivasa R, Yiwei C, Upadhyay L, et al (2001). Indole-3 carbinol (I3C) induced cell growth inhibition, G1 cell cycle arrest and apoptosis in Prostate cancer cells. Oncogene, 20, 2927-36. https://doi.org/10.1038/sj.onc.1204365
  28. Steven JT, Jackson, Singletary KW (2004). Sulforaphane: a naturally occurring mammary carcinoma mitotic inhibitor which disrupts tubulin polymerization. Carcinogenesis, 25, 219-27.
  29. Talalay P, Fahey JW (2001). Phytochemicals from cruciferous plant protect against Cancer by modulating Carcinogen metabolism. J Nutr, 131, 3027-33. https://doi.org/10.1093/jn/131.11.3027S
  30. Verkerk R, Dekker M, Jongen WM (2001). Post-Harvest increase of indolyl glucosinolatesin response to chopping and storage of Brassica vegetables. J Sci Food Agriculture, 81, 953-8. https://doi.org/10.1002/jsfa.854
  31. Yina X, Zhoua J, Jieb C, Xingc D, Zhanga Y (2004). Anticancer activity and mechanism of Scutellaria barbata extract on human lung cancer cell line A549. Life Sci, 75, 2233 -44. https://doi.org/10.1016/j.lfs.2004.05.015

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