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Probiotic Conjugated Linoleic Acid Mediated Apoptosis in Breast Cancer Cells by Downregulation of NF-κB

  • Kadirareddy, Rashmi Holur (Sri Padmavati Mahila Viswa Vidhyalayam) ;
  • GhantaVemuri, Sujana (Sri Padmavati Mahila Viswa Vidhyalayam) ;
  • Palempalli, Uma Maheswari Devi (Sri Padmavati Mahila Viswa Vidhyalayam)
  • Published : 2016.07.01

Abstract

Conjugated linoleic acid, a functional lipid, produced from Lactobacillus plantarum (LP-CLA), has been demonstrated to possess apoptotic activity. The anti-proliferative and apoptotic potential of LP-CLA was here evaluated in vitro using the MDA-MB-231 human breast cancer cell line as a model system. Proliferation of MDA-MB-231 cells was inhibited with increasing concentrations of LP-CLA with altered morphological features like cell detachment, rounding of cells and oligonucleosomal fragmentation of DNA. Flow cytometry confirmed the apoptotic potential of LP-CLA by ANNEXIN V/PI double staining. Furthermore, outcome results indicated that the apoptosis was mediated by downregulation of the NF-${\kappa}B$ pathway which in turn acted through proteasome degradation of $I{\kappa}B{\alpha}$, inhibition of p65 nuclear translocation, release of cytochrome-C from mitochondria and finally overexpression of Bax protein. Thus, conjugated linoleic acid, a natural product derived from probiotics, could therefore be a possible potential chemotherapeutic agent due to its apoptotic activity against estrogen receptor negative breast cancer cells.

Keywords

Conjugated linoleic acid;NF-${\kappa}B$;apoptosis;GSH;MDA-MB-231 cells;Lactobacillus plantarum

References

  1. Anderson ME (1985). Determination of glutathione and glutathione disulfide in biological samples. Methods Enzymol, 113, 548-55.
  2. Achenef M, Kadir AA, Othman F, Meng GY, Sazili AQ (2012). Cytotoxic effects of conjugated linoleic acids on human hepatoma cancer cells (HepG2). J Mol Pathophysiol 1, 43-8. https://doi.org/10.5455/jmp.20120127120302
  3. Chen FE, Huang DB, Chen YQ, Ghosh G (1998). Crystal structure of p50/p65 heterodimer of transcription factor NF-kappaB bound to DNA. Nature, 391, 410-413. https://doi.org/10.1038/34956
  4. Cho HJ, Kim WK, Kim EJ, Jung KC, Park S et al (2003). Conjugated linoleic acid inhibits cell proliferation and ErbB3 signaling in HT-29 human colon cell line. Am J Physiol Gastrointest Liver Physiol, 284, 996-1005. https://doi.org/10.1152/ajpgi.00347.2002
  5. El Roz A1, Bard JM, Huvelin JM, Nazih H (2013). The antiproliferative and pro-apoptotic effects of the trans9, trans11 conjugated linoleic acid isomer on MCF-7 breast cancer cells are associated with LXR activation. Prostaglandins Leukot Essent Fatty Acids, 88, 265-72. https://doi.org/10.1016/j.plefa.2012.12.006
  6. Fujimura M, Morita-Fujimura Y, Kawase M, et al (1999). Manganese superoxide dismutase mediates the early release of mitochondrial cytochrome C and subsequent DNA fragmentation after permanent focal cerebral ischemia in mice. J Neurosci, 19, 3414-22. https://doi.org/10.1523/JNEUROSCI.19-09-03414.1999
  7. Griffith OW (1980). Determination of glutathione and glutathione disulfide using glutathione reductase and 2-vinylpyridine. Anal Biochem, 106, 207-212. https://doi.org/10.1016/0003-2697(80)90139-6
  8. Hacker, H. M. Karin (2006). Regulation and function of IKK and IKK-related kinases. Sci STKE, 2006, 13.
  9. Heinze VM, Actis Dietary conjugated linoleic acid and longchain n-3 fatty acids in mammary and prostate cancer protection: a review. Int J Food Sci Nutr, 63, 66-78.
  10. Hosseini ES, Kermanshahi RK, Hosseinkhani S, Shojaosadati SA, Nazari M (2015). Conjugated linoleic acid production from various substrates by probiotic Lactobacillus plantarum. Ann Microbiol, 65, 27-32. https://doi.org/10.1007/s13213-014-0832-0
  11. Karin M, Delhase M (2000). The I kappa B kinase (IKK) and NF-kappa B: key elements of proinflammatory signalling. Semin Immunol, 12, 85-98. https://doi.org/10.1006/smim.2000.0210
  12. Kim KJ, Lee J, Park Y, Lee SH (2015). ATF3 mediates anticancer activity of trans-10, cis-12-conjugated linoleic acid in human colon cancer cells. Biomol Ther (Seoul). 23, 134-40. https://doi.org/10.4062/biomolther.2014.107
  13. Kucharczak J, Simmons MJ, Fan Y, Gelinas C (2003). To be, or not to be: NF-kappaB is the answer--role of Rel/NF-kappaB in the regulation of apoptosis. Oncogene, 22, 8961-82 https://doi.org/10.1038/sj.onc.1207230
  14. Law M, Corsino P, Parker NT, Law BK (2010). Identification of a small molecule inhibitor of serine 276 phosphorylation of the p65 subunit of NF-kappaB using in silico molecular docking. Cancer Lett, 291, 217-24. https://doi.org/10.1016/j.canlet.2009.10.015
  15. Liu P, Shen SR, Ruan H, et al (2011). Production of conjugated linoleic acids by Lactobacillus plantarum strains isolated from naturally fermented Chinese pickles. J Zhejiang Univ Sci B, 12, 923-30. https://doi.org/10.1631/jzus.B1100072
  16. Luhrmann A, Roy CR (2007). Coxiella burnetii inhibits activation of host cell apoptosis through a mechanism that involves preventing cytochrome c release from mitochondria. Infect Immun, 75, 5282-9. https://doi.org/10.1128/IAI.00863-07
  17. Mercurio F Manning AM (1999). NF-kappaB as a primary regulator of the stress response. Oncogene, 18, 6163-71. https://doi.org/10.1038/sj.onc.1203174
  18. Mihm SD, Galter, Droge W (2006). Modulation of transcription factor NF kappa B activity by intracellular glutathione levels and by variations of the extracellular cysteine supply. Faseb J, 9, 246-52.
  19. Nithya K, Santhi N, Kalaiselvi K (2009). Molecular docking of withanolides against the transcription factor nuclear factor-${\kappa}B$ (NF-kB) using glide. Adv Biotechnol, 9, 23-27.
  20. Navya A, Rashmi H, Vasavi T, Uma Maheswari Devi P (2014). Modulation of Pro-Inflammatory Genes by ${\alpha}$-Mangostin from Garcinia mangostana. Intl J Pharseu Sci Inventions, 3, 23-29.
  21. Ogawa J, Kishino S, Ando A, et al (2005). Production of conjugated fatty acids by lactic acid bacteria. J Biosci Bioeng, 100, 355-364. https://doi.org/10.1263/jbb.100.355
  22. Oosterhuis GJ, Mulder AB, Kalsbeek-Batenburg E, et al (2000). Measuring apoptosis in human spermatozoa: a biological assay for semen quality? Fertil Steril, 74, 245-50. https://doi.org/10.1016/S0015-0282(00)00623-3
  23. Palempalli UD, Gandhi U, Kalantari P, et al (2009). Gambogic acid covalently modifies IkappaB kinase-beta subunit to mediate suppression of lipopolysaccharide-induced activation of NF-kappaB in macrophages. Biochem J, 419, 401-9. https://doi.org/10.1042/BJ20081482
  24. Pardhasaradhi BV, Ali AM, Kumari AL, Reddanna P, Khar A (2003). Phycocyanin-mediated apoptosis in AK-5 tumor cells involves down-regulation of Bcl-2 and generation of ROS. Mol Cancer Ther, 2, 1165-70
  25. Pierre AS1, Minville-Walz M, Fevre C, Hichami A et al (2013) Trans-10, cis-12 conjugated linoleic acid induced cell death in human colon cancer cells through reactive oxygen speciesmediated ER stress. Biochim Biophys Acta, 1831, 759-68. https://doi.org/10.1016/j.bbalip.2013.01.005
  26. Rakib MA, Lee WS, Kim GS, et al (2013). Antiproliferative action of conjugated linoleic acid on human MCF-7 breast cancer cells mediated by enhancement of gap junctional intercellular communication through inactivation of NF- ${\kappa}B$. Evid Based Complement Alternat Med, 429393.
  27. Rothwarf DM, Karin M (1999). The NF-kappa B activation pathway: a paradigm in information transfer from membrane to nucleus. Sci STKE, RE1.
  28. Rungapamestry V, McMonagle J, Reynolds C, et al (2012). Inter-organ proteomic analysis reveals insights into the molecular mechanisms underlying the anti-diabetic effects of cis-9, trans-11-conjugated linoleic acid in ob/ob mice. Proteomics, 12, 461-76. https://doi.org/10.1002/pmic.201100312
  29. Sambrook J, F.E.F., Maniatis T (ed) (1989). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York
  30. Schreiber J, Jenner RG, Murray HL, et al (2006). Coordinated binding of NF-kappaB family members in the response of human cells to lipopolysaccharide. Proc Natl Acad Sci USA, 103, 5899-04. https://doi.org/10.1073/pnas.0510996103
  31. Shen W, Gaskins HR, McIntosh MK (2014). Influence of dietary fat on intestinal microbes, inflammation, barrier function and metabolic outcomes. J Nutr Biochem, 25, 270-80. https://doi.org/10.1016/j.jnutbio.2013.09.009
  32. Van Nieuwenhove CP, Oliszewski R, Gonzalez SN, Perez Chaia AB (2007). Conjugated linoleic acid conversion by dairy bacteria cultured in MRS broth and buffalo milk. Lett Appl Microbiol, 44, 467-74. https://doi.org/10.1111/j.1472-765X.2007.02135.x
  33. Vunta H, Davis F, Palempalli UD, et al (2007). The antiinflammatory effects of selenium are mediated through 15-deoxy-Delta12,14-prostaglandin J2 in macrophages. J Biol Chem, 282, 17964-73. https://doi.org/10.1074/jbc.M703075200
  34. Yang B, Chen H, Gu Z, et al (2014) Synthesis of conjugated linoleic acid by the linoleate isomerase complex in foodderived lactobacilli. J Appl Microbiol, 117, 430-9. https://doi.org/10.1111/jam.12524
  35. Zeng Z, Lin J, Gong D (2009). Identification of lactic acid bacterial strains with high conjugated linoleic acid-producing ability from natural sauerkraut fermentations. J Food Sci, 74, 154-158.
  36. Zhang HF, Wu D, Du JK, Zhang Y, Su YM (2014). Inhibitory effects of phenolic alkaloids of Menispermum dauricum on gastric cancer in vivo. Asian Pac J Cancer Prev, 15, 10825-30.