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Non steroidal anti-inflammatory drugs modulate the physicochemical properties of plasma membrane in experimental colorectal cancer: a fluorescence spectroscopic study

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Abstract

According to “fluid-mosaic model,” plasma membrane is a bilayer constituted by phospholipids which regulates the various cellular activities governed by many proteins and enzymes. Any chemical, biochemical, or physical factor has to interact with the bilayer in order to regulate the cellular metabolism where various physicochemical properties of membrane, i.e., polarization, fluidity, electrostatic potential, and phase state may get affected. In this study, we have observed the in vivo effects of a pro-carcinogen 1,2-dimethylhydrazine dihydrochloride (DMH) and the two non steroidal anti-inflammatory drugs (NSAIDs); sulindac and celecoxib on various properties of the plasma membrane of colonocytes, i.e., electric potential, fluidity, anisotropy, microviscosity, lateral diffusion, and phase state in the experimentally induced colorectal cancer. A number of fluorescence probes were utilized like membrane fluidity and anisotropy by 1,6-diphenyl-1,3,5-hexatriene, membrane microviscosity by Pyrene, membrane electric potential by merocyanine 540, lateral diffusion by N-NBD-PE, and phase state by Laurdan. It is observed that membrane phospholipids are less densely packed and therefore, the membrane is more fluid in case of carcinogenesis produced by DMH than control. But NSAIDs are effective in reverting back the membrane toward normal state when co-administered with DMH. The membrane becomes less fluid, composed of low electric potential phospholipids whose lateral diffusion is being prohibited and the membrane stays mostly in relative gel phase. It may be stated that sulindac and celecoxib, the two NSAIDs may exert their anti-neoplastic role in colorectal cancer via modifying the physicochemical properties of the membranes.

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References

  1. Parasassi T, Stasio GD, d’Ubaldo A, Gratton E (1990) Phase fluctuation in phospholipid membranes revealed by Laurdan fluorescence. Biophys J 57:1179–1186

    Article  PubMed  CAS  Google Scholar 

  2. Koivusalo M, Alvesalo J, Virtanen JA, Somerharju P (2004) Partitioning of pyrene-labeled phospho- and sphingolipids between ordered and disordered bilayer domains. Biophys J 86:923–935

    Article  PubMed  CAS  Google Scholar 

  3. Cantor RS (1999) Lipid composition and the lateral pressure profile in bilayers. Biophys J 76:2625–2639

    Article  PubMed  CAS  Google Scholar 

  4. Parasassi T, Stasio GD, Ravagnan G, Rusch RM, Gratton E (1991) Quantitation of lipid phases in phospholipid vesicles by the generalized polarization of Laurdan fluorescence. Biophys J 60:179–189

    Article  PubMed  CAS  Google Scholar 

  5. Lúcio M, Ferreira H, Lima JLFC, Matos C, de Castro B, Reis S (2004) Influence of some anti-inflammatory drugs in membrane fluidity studied by fluorescence anisotropy measurements. Phys Chem Chem Phys 6:1493–1498. doi:10.1039/b314551b

    Article  Google Scholar 

  6. Maity P, Bindu S, Dey S, Goyal M, Alam A, Pal C, Mitra K, Bandyopadhyay U (2009) Indomethacin, a non-steroidal anti-inflammatory drug, develops gastropathy by inducing reactive oxygen species-mediated mitochondrial pathology and associated apoptosis in gastric mucosa. J Biol Chem 284:3058–3068. doi:10.1074/jbc.M805329200

    Article  PubMed  CAS  Google Scholar 

  7. Lamont EB, Dias LE, Lauderdale DS (2007) NSAIDS and colorectal cancer risk: do administrative data support a chemopreventive effect? J Gen Intern Med 22:1166–1171. doi:10.1007/s11606-007-0256-7

    Article  PubMed  Google Scholar 

  8. Zhou Y, Hancock JF, Lichtenberger LM (2010) The nonsteroidal anti-inflammatory drug indomethacin induces heterogeneity in lipid membranes: potential implication for its diverse biological action. PLoS ONE 5:e8811. doi:10.1371/journal.pone.000881

    Article  PubMed  Google Scholar 

  9. Kanwar SS, Vaipei K, Nehru B, Sanyal SN (2008) Antioxidative effects of non-steroidal anti-inflammatory drugs during the initiation stages of experimental colon carcinogenesis in rat. J Environ Pathol Toxicol Oncol 27:89–100

    PubMed  CAS  Google Scholar 

  10. Vaish V, Tanwar L, Sanyal SN (2010) The role of NF-κB and PPARγ in experimentally induced colorectal cancer and chemoprevention by cyclooxygenase-2 inhibitors. Tumour Biol 31:427–436. doi:10.1007/s13277-010-0051-7

    Article  PubMed  CAS  Google Scholar 

  11. Brown WA, Skinner SA, Malcontenti-Wilson C, Vogiagis D, O’Brien PE (2001) Non-steroidal anti-inflammatory drugs with activity against either cyclooxygenase 1 or cyclooxygenase 2 inhibit colorectal cancer in a DMH rodent model by inducing apoptosis and inhibiting cell proliferation. Gut 48:660–666

    Article  PubMed  CAS  Google Scholar 

  12. Vaish V, Sanyal SN (2011) Chemopreventive effects of NSAIDs on cytokines and transcription factors at the early stages of colorectal cancer. Pharmacol Rep 63(5) (in press)

  13. Mouille B, Robert V, Blachier F (2004) Adaptive increase of ornithine production and decrease of ammonia metabolism in rat colonocytes and hyperproteic diet ingestion. Am J Physiol Gastrointest Liver Physiol 287:G344–G351

    Article  PubMed  CAS  Google Scholar 

  14. Roediger WE, Truelove C (1979) Method of preparing isolated colonic epithelial cells (colonocytes) for metabolic studies. Gut 20:484–488

    Article  PubMed  CAS  Google Scholar 

  15. Siboni G, Rothmann C, Ehrenberg B, Malik Z (2001) Spectral imaging of MC540 during murine and human colon carcinoma cell differentiation. J Histochem Cytochem 49:147–153

    Article  PubMed  CAS  Google Scholar 

  16. Baumber J, Meyers SA (2006) Changes in membrane lipid order with capacitation in rhesus macaque (Macaca mulatta) spermatozoa. J Androl 27:578–587. doi:10.2164/jandrol.05135

    Article  PubMed  CAS  Google Scholar 

  17. Nichols JW, Pagano RE (1981) Kinetics of soluble lipid monomer diffusion between vesicles. Biochemistry 20:2783–2789

    Article  PubMed  CAS  Google Scholar 

  18. Hoekstra D (1982) Fluorescence method for measuring the kinetics of Ca2+-induced phase separations in phosphatidylserine-containing lipid vesicles. Biochemistry 21:1055–1061

    Article  PubMed  CAS  Google Scholar 

  19. Ambrosini A, Zolese G, Wozniak M, Genga D, Boscaro M, Mantero F, Balerica G (2003) Idiopathic infertility: susceptibility of spermatozoa to in vitro capacitation, in the presence and the absence of palmitylethanolamide (a homologue of anandamide), is strongly correlated with membrane polarity studies by Laurdan fluorescence. Mol Human Reprod 9:381–388

    Article  CAS  Google Scholar 

  20. Parasassi T, Ravagnan G, Rusch RM, Gratton E (1993) Modulation and dynamics of phase properties in phospholipid mixtures detected by Laurdan fluorescence. Photochem Photobiol 57:403–410

    Article  PubMed  CAS  Google Scholar 

  21. Shinitzky M, Barenholz Y (1974) Dynamics of the hydrocarbon layer in liposomes of lecithin and sphingomyelin containing dicetylphosphate. J Biol Chem 249:2652–2657

    PubMed  CAS  Google Scholar 

  22. Pottel H, van der Meer W, Herreman W (1983) Correlation between the order parameter and the steady state fluorescence anisotropy of 1,6 diphenyl-1,3,5-hexatriene and an evaluation of membrane fluidity. Biochim Biophys Acta 730:181–186

    Article  CAS  Google Scholar 

  23. Massey V, Williums CH (1965) On the reaction mechanism of yeast glutathione reductase. J Biol Chem 240:4470–4475

    PubMed  CAS  Google Scholar 

  24. Vanderkooi JM, Callis JB (1974) Pyrene. Probe of lateral diffusion in the hydrophobic region of membranes. Biochemistry 13:4000–4006

    Article  PubMed  CAS  Google Scholar 

  25. Levine L (2006) Cyclooxygenase expression is not required for release of arachidonic acid from cells by some nonsteroidal anti-inflammatory drugs and cancer preventive agents. BMC Pharmacol 6:7

    Article  PubMed  Google Scholar 

  26. Kaur J, Sanyal SN (2010) Alterations in membrane fluidity and dynamics in experimental colon cancer and its chemoprevention by diclofenac. Mol Cell Biochem 341:99–108

    Article  PubMed  CAS  Google Scholar 

  27. Kalyanaraman B, Feix JB, Siebert F, Thomas JP, Girotti AW (1987) Photodynamic action of merocyanine 540 on artificial and natural cell membranes: Involvement of singlet molecular oxygen. Proc Natl Acad Sci USA 84:2999–3003

    Article  PubMed  CAS  Google Scholar 

  28. McEvoy L, Schiegel RA, Williamson P, Buono BJD (1988) Merocyanine 540 as a flow cytometric probe of membrane lipid organization in leukocytes. J Leukoc Biol 44:337–344

    PubMed  CAS  Google Scholar 

  29. John K, Schreiber S, Kubelt J, Herrmann A, Müller P (2002) Transbilayer movement of phospholipids at the main phase transition of lipid membranes: implications for rapid flip-flop in biological membranes. Biophys J 83:3315–3323

    Article  PubMed  CAS  Google Scholar 

  30. Bonnafous P, Stegmann T (2000) Membrane perturbation and fusion pore formation in influenza hemagglutinin-mediated membrane fusion. J Biol Chem 275:6160–6166

    Article  PubMed  CAS  Google Scholar 

  31. Weber G, Farris FJ (1979) Synthesis and spectral properties of a hydrophobic fluorescent probe: 6-propionyl-2-(dimethylamino)naphthalene. Biochemistry 18:3075–3078

    Article  PubMed  CAS  Google Scholar 

  32. Weber P, Wagner M, Schneckenburger H (2006) Microfluorometry of cell membrane dynamics. CytometryA 69A:185–188. doi:10.1002/cyto.a.20233

    Article  Google Scholar 

  33. Parasassi T, Stefano MD, Loiero M, Ravagnan G, Gratton E (1994) Cholesterol modifies water concentration and dynamics in phospholipid bilayers: a fluorescence study using Laurdan probe. Biophys J 66:763–768

    Article  PubMed  CAS  Google Scholar 

  34. Parasassi T, Stefano MD, Loiero M, Ravagnan G, Gratton E (1994) Influence of cholesterol on phospholipid bilayers phase domains as detected by Laurdan fluorescence. Biophys J 66:120–132

    Article  PubMed  CAS  Google Scholar 

  35. Parasassi T, Gratton E, Yu WM, Wilson P, Levi M (1997) Two-photon fluorescence microscopy of Laurdan generalized polarization domains in model and natural membranes. Biophys J 72:2413–2429

    Article  PubMed  CAS  Google Scholar 

  36. Dale RE, Chen LA, Brand L (1971) Rotational relaxation of the “microviscosity” probe diphenylhexatriene in paraffin oil and egg lecithin vesicles. J Biol Chem 252:1500–1510

    Google Scholar 

  37. Barenholz Y, Cohen T, Haas E, Ottolenghi M (1996) Lateral organization of pyrene-labeled lipids in bilayers as determined from the deviation from equilibrium between pyrene monomers and excimers. J Biol Chem 271:3085–3090

    Article  PubMed  CAS  Google Scholar 

  38. Wilson-Ashworth HA, Bahm Q, Erickson J, Shinkle A, Vu MP, Woodbury D, Bell JD (2006) Differential detection of phospholipid fluidity, order and spacing by fluorescence spectroscopy of bis-pyrene, prodan, nystatin, and merocyanine 540. Biophys J 91:4091–5001

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

Financial assistance from the Department of Science and Technology, Govt. of India (SR/SO/BB-05/2008) is gratefully acknowledged.

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Correspondence to Sankar Nath Sanyal.

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Vaish, V., Sanyal, S.N. Non steroidal anti-inflammatory drugs modulate the physicochemical properties of plasma membrane in experimental colorectal cancer: a fluorescence spectroscopic study. Mol Cell Biochem 358, 161–171 (2011). https://doi.org/10.1007/s11010-011-0931-1

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  • DOI: https://doi.org/10.1007/s11010-011-0931-1

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