Effect of the microbial lipopeptide on tumor
cell lines: apoptosis induced by disturbing the fatty acid composition
of cell membrane

doi:10.1007/s13238-010-0072-4

Protein Cell

2010, Vol. 1

Issue (6) : 584-594 https://doi.org/10.1007/s13238-010-0072-4 PMID: 21204010

Research articles

Effect of the microbial lipopeptide on tumor cell lines: apoptosis induced by disturbing the fatty acid composition of cell membrane

Xiangyang Liu¹,Xinyi Tao²,Aihua Zou²,Shizhong Yang²,Bozhong Mu²,Lixin Zhang³,

1.State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai 200237, China；€Institute of Microbiology, Chinese Academy of Sciences, Beijing 100190, China; 2.State Key Laboratory of Bioreactor Engineering and Institute of Applied Chemistry, East China University of Science and Technology, Shanghai 200237, China; 3.€Institute of Microbiology, Chinese Academy of Sciences, Beijing 100190, China;

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Abstract Microbial lipopeptides play an important role in apoptosis induction of tumor cells. However, there is little knowledge about the relationship between apoptosis induction and membrane fatty acids. The present study focused on the effects of lipopeptides produced by Bacillus subtilis HSO121 on Bcap-37 cell lines. 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl (MTT) colorimetric assay and surface tension measurements, showed that the critical micelle concentration (CMC) was a critical level for the inhibitory activity of lipopeptides on the growth of Bcap-37 cells. Under the CMC, the order of least to greatest cytotoxicity effect on cancer cell lines by lipopeptides is C₁₃-lipopeptide<C₁₄-lipopepitde<C₁₅-lipopeptide. Above CMC, all lipopeptides directly exert cytolytic activity. The flow cytometric analysis and Hoechst33258 staining experiments confirmed the apoptosis of Bcap-37 cell lines induced by lipopeptides in a dose-dependent manner. This apoptosis was associated with a significant decrease of the unsaturated degree of the cellular fatty acids of Bcap-37 cell lines due to the changes in the cellular fatty acids composition induced by the lipopeptide treatment. These results indicated that disturbance of the cellular fatty acid composition of breast cancer cell lines were related to in the cell apoptosis. Furthermore, significant difference in IC₅₀ values of tumor cells and normal cell showed that the lipopeptide exerted selective cytotoxicity on the cancer cells. Thus HSO121 lipopeptides may have potential applications as an anticancer leads.

Keywords Bacillus subtilis lipopeptide antitumor activity membrane fatty acid apoptosis

Issue Date: 01 June 2010

Cite this article:

Xinyi Tao,Xiangyang Liu,Aihua Zou, et al. Effect of the microbial lipopeptide on tumor cell lines: apoptosis induced by disturbing the fatty acid composition of cell membrane[J]. Protein Cell, 2010, 1(6): 584-594.

URL:

https://academic.hep.com.cn/pac/EN/10.1007/s13238-010-0072-4
https://academic.hep.com.cn/pac/EN/Y2010/V1/I6/584

	Ando, H., Wen, Z.M., Kim, H.Y., Valencia, J.C., Costin, G.E., Watabe, H., Yasumoto, K., Niki, Y., Kondoh, H., Ichihashi, M., et al. (2006). Intracellular composition of fatty acid affectsthe processing and function of tyrosinase through the ubiquitin-proteasomepathway. Biochem J394, 43–50. doi: 10.1042/BJ20051419
	Arima, K., Kakinuma, A., and Tamura, G. (1968). Surfactin, a crystallinepeptidelipid surfactant produced by Bacillussubtilis: isolation, characterization and its inhibitionof fibrin clot formation. Biochem BiophysRes Commun31, 488–494. doi: 10.1016/0006-291X(68)90503-2
	Bligh, E.G., and Dyer, W.J. (1959). A rapidmethod of total lipid extraction and purification. Can J Biochem Physiol37, 911–917.
	Bonmatin, J.M., Laprévote, O., and Peypoux, F. (2003). Diversity amongmicrobial cyclic lipopeptides: iturins and surfactins. Activity-structurerelationships to design new bioactive agents. Comb Chem High Throughput Screen6, 541–556.
	Bortolato, M., Besson, F., and Roux, B. (1997). Inhibition of alkalinephosphatase by surfactin, a natural chelating lipopeptide from Bacillus subtilis. Biotechnol Lett19, 433–435. doi: 10.1023/A:1018387909310
	Cao, X., Wang, A.H., Jiao, R.Z., Wang, C.L., Mao, D.Z., Yan, L., and Zeng, B. (2009a). Surfactininduces apoptosis and G(2)/M arrest in human breast cancer MCF-7 cellsthrough cell cycle factor regulation. CellBiochem Biophys55, 163–171. doi: 10.1007/s12013-009-9065-4
	Cao, X.H., Liao, Z.Y., Wang, C.L., Cai, P., Yang, W.Y., Lu, M.F., and Huang, G.W. (2009b). Purificationand antitumour activity of a lipopeptide biosurfactant produced by Bacillus natto TK-1. Biotechnol Appl Biochem52, 97–106. doi: 10.1042/BA20070227
	Cao, X.H., Liao, Z.Y., Wang, C.L., Yang, W.Y., and Lu, M.F. (2009c). Evaluationof a lipopeptide biosurfactant from Bacillusnatto Tk-1 as a potential source of anti-adhesive. Antimicrobialand Antitumor Activities. Braz J Microbiol40, 373–379. doi: 10.1590/S1517-83822009000200030
	Cao, X.H., Wang, A.H., Wang, C.L., Mao, D.Z., Lu, M.F., Cui, Y.Q., and Jiao, R.Z. (2010). Surfactininduces apoptosis in human breast cancer MCF-7 cells through a ROS/JNK-mediatedmitochondrial/caspase pathway. Chem BiolInteract183, 357–362. doi: 10.1016/j.cbi.2009.11.027
	Carrillo, C., Teruel, J.A., Aranda, F.J., and Ortiz, A. (2003). Molecular mechanism of membrane permeabilization bythe peptide antibiotic surfactin. BiochimBiophys Acta1611, 91–97. doi: 10.1016/S0005-2736(03)00029-4
	Dai, R.J., Phillips, R.A., Zhang, Y., Khan, D., Crasta, O., and Ahmed, S.A. (2008). Suppression of LPS-induced Interferon-gamma and nitricoxide in splenic lymphocytes by select estrogen-regulated microRNAs:a novel mechanism of immune modulation. Blood112, 4591–4597. doi: 10.1182/blood-2008-04-152488
	Deleu, M., Bouffioux, O., Razafindralambo, H., Paquot, M., Hbid, C., Thonart, P., Jacques, P., and Brasseur, R. (2003). Interactionof surfactin with membranes: a computational approach. Langmuir19, 3377–3385. doi: 10.1021/la026543z
	Dufour, S., Deleu, M., Nott, K., Wathelet, B., Thonart, P., and Paquot, M. (2005). Hemolytic activityof new linear surfactin analogs in relation to their physico-chemicalproperties. Biochim Biophys Acta1726, 87–95.
	Eeman, M., Berquand, A., Dufrêne, Y.F., Paquot, M., Dufour, S., and Deleu, M. (2006). Penetration of surfactininto phospholipid monolayers: nanoscale interfacial organization. Langmuir22, 11337–11345. doi: 10.1021/la061969p
	Elstein, K.H., Thomas, D.J., and Zucker, R.M. (1995). Factors affectingflow cytometric detection of apoptotic nuclei by DNA analysis. Cytometry21, 170–176. doi: 10.1002/cyto.990210209
	Geiser, F., McAllan, B.M., and Kenagy, G.J. (1994). The degree of dietaryfatty acid unsaturation affects torpor patterns and lipid compositionof a hibernator. J Comp Physiol B164, 299–305. doi: 10.1007/BF00346446
	Geyer, R.P., Bennett, A., and Rohr, A. (1962). Fatty acids of thetriglycerides and phospholipids of HeLa cells and strain L fibroblasts. J Lipid Res3, 80–83.
	Gong, J., Traganos, F., and Darzynkiewicz, Z. (1994). A selectiveprocedure for DNA extraction from apoptotic cells applicable for gelelectrophoresis and flow cytometry. AnalBiochem218, 314–319. doi: 10.1006/abio.1994.1184
	Gorczyca, W., Gong, J., and Darzynkiewicz, Z. (1993). Detectionof DNA strand breaks in individual apoptotic cells by the in situterminal deoxynucleotidyl transferase and nick translation assays. Cancer Res53, 1945–1951.
	Grau, A., Gómez Fernández, J.C., Peypoux, F., and Ortiz, A. (1999). A study on the interactionsof surfactin with phospholipid vesicles. Biochim Biophys Acta1418, 307–319. doi: 10.1016/S0005-2736(99)00039-5
	Haddad, N.I.A., Liu, X.Y., Yang, S.Z., and Mu, B.Z. (2008). Surfactin isoforms from Bacillussubtilis HSO121: separation and characterization. Protein Pept Lett15, 265–269. doi: 10.2174/092986608783744225
	Hagve, T.A. (1988). Effects of unsaturated fatty acids on cell membranefunctions. Scand J Clin Lab Invest48, 381–388.
	Heerklotz, H., and Seelig, J. (2001). Detergent-likeaction of the antibiotic peptide surfactin on lipid membranes. Biophys J81, 1547–1554.
	Heerklotz, H., and Seelig, J. (2007). Leakageand lysis of lipid membranes induced by the lipopeptide surfactin. Eur Biophys J36, 305–314. doi: 10.1007/s00249-006-0091-5
	Howlett, N.G., and Avery, S.V. (1997). Relationshipbetween cadmium sensitivity and degree of plasma membrane fatty acidunsaturation in Saccharomyces cerevisiae. Appl Microbiol Biotechnol48, 539–545. doi: 10.1007/s002530051093
	Huang, X.Q., Lu, Z.X., Zhao, H.Z., Bie, X.M., Xia, L.F., and Yang, S.J. (2006). Antiviral activity of antimicrobial lipopeptide from Bacillus subtilis fmbj against PseudorabiesVirus, Porcine Parvovirus, Newcastle Disease Virus and InfectiousBursal Disease Virus in Vitro. Int J PeptRes Ther12, 373–377. doi: 10.1007/s10989-006-9041-4
	Kameda, Y., Oira, S., Matsui, K., Kanatomo, S., and Hase, T. (1974). Antitumoractivity of bacillus natto. V. Isolation and characterization of surfactinin the culture medium of Bacillus natto KMD 2311. Chem Pharm Bull (Tokyo)22, 938–944.
	Kikuchi, T., and Hasumi, K. (2002). Enhancementof plasminogen activation by surfactin C: augmentation of fibrinolysisin vitro and in vivo. Biochim Biophys Acta1596, 234–245.
	Kim, S.Y., Kim, J.Y., Kim, S.H., Bae, H.J., Yi, H., Yoon, S.H., Koo, B.S., Kwon, M., Cho, J.Y., Lee, C.E., et al. (2007). Surfactinfrom Bacillus subtilis displaysanti-proliferative effect via apoptosis induction, cell cycle arrestand survival signaling suppression. FEBSLett581, 865–871. doi: 10.1016/j.febslet.2007.01.059
	Kuo, M.T. (2009). Redox regulation of multidrug resistance in cancer chemotherapy:molecular mechanisms and therapeutic opportunities. Antioxid Redox Signal11, 99–133. doi: 10.1089/ars.2008.2095
	Liu, X.Y., Haddad, N.I.A., Yang, S.Z., and Mu, B.Z. (2007). Structural characterization of eight cyclic lipopeptidesproduced by Bacillus subtilis HSO121. Protein Pept Lett14, 766–773. doi: 10.2174/092986607781483642
	Liu, X.Y., Yang, S.Z., and Mu, B.Z. (2008). Isolation and characterizationof a C₁₂-lipopeptide produced by Bacillus subtilis HSO 121. J Pept Sci14, 864–875. doi: 10.1002/psc.1017
	Marsh, S., and McLeod, H.L. (2007). Pharmacogeneticsand oncology treatment for breast cancer. Expert Opin Pharmacother8, 119–127. doi: 10.1517/14656566.8.2.119
	Menendez, J.A., Vellon, L., and Lupu, R. (2005). Targeting fattyacid synthase-driven lipid rafts: a novel strategy to overcome trastuzumabresistance in breast cancer cells. MedHypotheses64, 997–1001. doi: 10.1016/j.mehy.2004.09.027
	Michel, V., and Bakovic, M. (2007). Lipidrafts in health and disease. Biol Cell99, 129–140. doi: 10.1042/BC20060051
	Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays. J Immunol Methods65, 55–63. doi: 10.1016/0022-1759(83)90303-4
	Pelz, K., Hopfener, K., Wiedmann-Al-Ahmad, M., Jahnke, H., Wittmer, A., and Otten, J.E. (2006). Differences in thefatty acid composition of KB-cells and gingival keratinocytes is culturemedium additive dependent. Biomed Chromatogr20, 870–880. doi: 10.1002/bmc.610
	Qi, L.F., Xu, Z.R., and Chen, M.L. (2007). In vitro and invivo suppression of hepatocellular carcinoma growth by chitosan nanoparticles. Eur J Cancer43, 184–193. doi: 10.1016/j.ejca.2006.08.029
	Rakheja, D., Kapur, P., Hoang, M.P., Roy, L.C., and Bennett, M.J. (2005). Increasedratio of saturated to unsaturated C18 fatty acids in colonic adenocarcinoma:implications for cryotherapy and lipid raft function. Med Hypotheses65, 1120–1123. doi: 10.1016/j.mehy.2005.05.045
	Savage, P.B., Li, C., Taotafa, U., Ding, B., and Guan, Q. (2002). Antibacterial propertiesof cationic steroid antibiotics. FEMS MicrobiolLett217, 1–7. doi: 10.1111/j.1574-6968.2002.tb11448.x
	Schlager, S.I., Ohanian, S.H., and Borsos, T. (1978). Correlation betweenthe ability of tumor cells to incorporate specific fatty acids andtheir sensitivity to killing by a specific antibody plus guinea pigcomplement. J Natl Cancer Inst61, 931–934.
	Sheppard, J.D., Jumarie, C., Cooper, D.G., and Laprade, R. (1991). Ionic channels induced by surfactin in planar lipidbilayer membranes. Biochim Biophys Acta1064, 13–23. doi: 10.1016/0005-2736(91)90406-X
	Singletary, S.E. (2008). Breast cancer management: the road to today. Cancer113, 1844–1849. doi: 10.1002/cncr.23649
	Vaara, M. (1992). Agents that increase the permeability of the outer membrane. Microbiol Rev56, 395–411.
	Vollenbroich, D., Pauli, G., ?zel, M., and Vater, J. (1997). Antimycoplasma properties and application in cell cultureof surfactin, a lipopeptide antibiotic from Bacillus subtilis. Appl EnvironMicrobiol63, 44–49.
	Walker, P.R., Kwast-Welfeld, J., Gourdeau, H., Leblanc, J., Neugebauer, W., and Sikorska, M. (1993). Relationship betweenapoptosis and the cell cycle in lymphocytes: roles of protein kinaseC, tyrosine phosphorylation, and AP1. ExpCell Res207, 142–151. doi: 10.1006/excr.1993.1173
	Wang, C.L., Ng, T.B., Yuan, F., Liu, Z.K., and Liu, F. (2007). Inductionof apoptosis in human leukemia K562 cells by cyclic lipopeptide from Bacillus subtilis natto T-2. Peptides28, 1344–1350. doi: 10.1016/j.peptides.2007.06.014
	Ye, C.L., Liu, J.W., Wei, D.Z., Lu, Y.H., and Qian, F. (2004). In vitroanti-tumor activity of 2',4'-dihydroxy-6'-methoxy-3',5'-dimethylchalconeagainst six established human cancer cell lines. Pharmacol Res50, 505–510. doi: 10.1016/j.phrs.2004.05.004

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