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Frontiers of Chemical Science and Engineering

ISSN 2095-0179

ISSN 2095-0187(Online)

CN 11-5981/TQ

邮发代号 80-969

2019 Impact Factor: 3.552

Frontiers of Chemical Science and Engineering  2019, Vol. 13 Issue (1): 98-107   https://doi.org/10.1007/s11705-018-1775-y
  本期目录
Insight into the role of cholesterol in modulation of morphology and mechanical properties of CHO-K1 cells: An in situ AFM study
Lei Zhang1,2, Lisha Zhao3, Ping-Kai Ouyang2, Pu Chen1,2()
1. Department of Chemical Engineering and Waterloo Institute for Nanotechnology, University of Waterloo, Ontario N2L 3G1, Canada
2. College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing 211816, China
3. Department of Agricultural, Food and Nutritional Science, University of Alberta, Alberta T6G 2P5, Canada
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Abstract

Cholesterol plays a significant role in the organization of lipids and modulation of membrane dynamics in mammalian cells. However, the effect of cholesterol depletion on the eukaryotic cell membranes seems controversial. In this study, the effects of cholesterol on the topography and mechanical behaviors of CHO-K1 cells with manipulated membrane cholesterol contents were investigated by atomic force microscopy (AFM) technique. Here, we found that the depletion of cholesterol in cell membranes could increase the membrane stiffness, reduce the cell height as well as promote cell retraction and detachment from the surface, whereas the cholesterol restoration could reverse the effect of cholesterol depletion on the membrane stiffness. Increased methyl-β-cyclodextrin levels and incubation time could significantly increase Young’s modulus and degree of stiffing on cell membrane and cytoskeleton. This research demonstratede importance of cholesterol in regulating the dynamics of cytoskeleton-mediated processes. AFM technique offers excellent advantages in the dynamic monitoring of the change in membranes mechanical properties and behaviors during the imaging process. This promising technology can be utilized in studying the membrane properties and elucidating the underlying mechanism of distinct cells in the near-native environment.

Key wordscholesterol    methyl-β-cyclodextrin    atomic force microscopy    Young’s modulus    CHO-K1 cell
收稿日期: 2018-05-23      出版日期: 2019-02-25
Corresponding Author(s): Pu Chen   
 引用本文:   
. [J]. Frontiers of Chemical Science and Engineering, 2019, 13(1): 98-107.
Lei Zhang, Lisha Zhao, Ping-Kai Ouyang, Pu Chen. Insight into the role of cholesterol in modulation of morphology and mechanical properties of CHO-K1 cells: An in situ AFM study. Front. Chem. Sci. Eng., 2019, 13(1): 98-107.
 链接本文:  
https://academic.hep.com.cn/fcse/CN/10.1007/s11705-018-1775-y
https://academic.hep.com.cn/fcse/CN/Y2019/V13/I1/98
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1 Lde Oliveira Andrade. Understanding the role of cholesterol in cellular biomechanics and regulation of vesicular trafficking: The power of imaging. Biomedical Spectroscopy and Imaging, 2016, 5(s1): S101–S117
https://doi.org/10.3233/BSI-160157
2 EEvangelisti, C Cecchi, RCascella, CSgromo, MBecatti, C MDobson, FChiti, MStefani. Membrane lipid composition and its physicochemical properties define cell vulnerability to aberrant protein oligomers. Journal of Cell Science, 2012, 125(10): 2416–2427
https://doi.org/10.1242/jcs.098434
3 LRedondo-Morata, M I Giannotti, F Sanz. Influence of cholesterol on the phase transition of lipid bilayers: A temperature-controlled force spectroscopy study. Langmuir, 2012, 28(35): 12851–12860
https://doi.org/10.1021/la302620t
4 LZhao, F Temelli. Preparation of liposomes using supercritical carbon dioxide via depressurization of the supercritical phase. Journal of Food Engineering, 2015, 158: 104–112
https://doi.org/10.1016/j.jfoodeng.2015.03.004
5 AMagarkar, V Dhawan, PKallinteri, TViitala, MElmowafy, TRog, A Bunker. Cholesterol level affects surface charge of lipid membranes in saline solution. Scientific Reports, 2014, 4: 2045–2322
6 LZhao, F Temelli, J MCurtis, LChen. Encapsulation of lutein in liposomes using supercritical carbon dioxide. Food Research International, 2017, 100: 168–179
https://doi.org/10.1016/j.foodres.2017.06.055
7 Fde Meyer, B Smit. Effect of cholesterol on the structure of a phospholipid bilayer. Proceedings of the National Academy of Sciences of the United States of America, 2009, 106(10): 3654–3658
https://doi.org/10.1073/pnas.0809959106
8 WSułkowski, DPentak, KNowak, ASułkowska. The influence of temperature, cholesterol content and pH on liposome stability. Journal of Molecular Structure, 2005, 744: 737–747
https://doi.org/10.1016/j.molstruc.2004.11.075
9 LZhao, F Temelli, J MCurtis, LChen. Preparation of liposomes using supercritical carbon dioxide technology: Effects of phospholipids and sterols. Food Research International, 2015, 77: 63–72
https://doi.org/10.1016/j.foodres.2015.07.006
10 LZhao, F Temelli. Preparation of liposomes using a modified supercritical process via depressurization of liquid phase. Journal of Supercritical Fluids, 2015, 100: 110–120
https://doi.org/10.1016/j.supflu.2015.02.022
11 NKhatibzadeh, A A Spector, W E Brownell, B Anvari. Effects of plasma membrane cholesterol level and cytoskeleton F-actin on cell protrusion mechanics. PLoS One, 2013, 8(2): e57147
https://doi.org/10.1371/journal.pone.0057147
12 SMañes, C Martínez-A. Cholesterol domains regulate the actin cytoskeleton at the leading edge of moving cells. Trends in Cell Biology, 2004, 14(6): 275–278
https://doi.org/10.1016/j.tcb.2004.04.008
13 MSun, N Northup, FMarga, THuber, F JByfield, ILevitan, GForgacs. The effect of cellular cholesterol on membrane-cytoskeleton adhesion. Journal of Cell Science, 2007, 120(13): 2223–2231
https://doi.org/10.1242/jcs.001370
14 L LNorman, R J Oetama, M Dembo, FByfield, D AHammer, ILevitan, HAranda-Espinoza. Modification of cellular cholesterol content affects traction force, adhesion and cell spreading. Cellular and Molecular Bioengineering, 2010, 3(2): 151–162
https://doi.org/10.1007/s12195-010-0119-x
15 Y TYang, J D Liao, C C K Lin, C T Chang, S H Wang, M S Ju. Characterization of cholesterol-depleted or-restored cell membranes by depth-sensing nano-indentation. Soft Matter, 2012, 8(3): 682–687
https://doi.org/10.1039/C1SM06180A
16 PKilbride, H J Woodward, K B Tan, N T Thanh, K E Chu, S Minogue, M GWaugh. Modeling the effects of cyclodextrin on intracellular membrane vesicles from Cos-7 cells prepared by sonication and carbonate treatment. PeerJ, 2015, 3: e1351
https://doi.org/10.7717/peerj.1351
17 RZidovetzki, I Levitan. Use of cyclodextrins to manipulate plasma membrane cholesterol content: Evidence, misconceptions and control strategies. Biochimica et Biophysica Acta (BBA)— Biomembranes, 2007, 1768(6): 1311–1324
https://doi.org/10.1016/j.bbamem.2007.03.026
18 AChristian, M Haynes, MPhillips, GRothblat. Use of cyclodextrins for manipulating cellular cholesterol content. Journal of Lipid Research, 1997, 38(11): 2264–2272
19 V GRomanenko, YFang, F Byfield, A JTravis, C AVandenberg, G HRothblat, ILevitan. Cholesterol sensitivity and lipid raft targeting of Kir2. 1 channels. Biophysical Journal, 2004, 87(6): 3850–3861
https://doi.org/10.1529/biophysj.104.043273
20 SMahammad, I Parmryd. Cholesterol depletion using methyl--cyclodextrin. Methods in Membrane Lipids, 2015: 91–102
21 V GRomanenko, G HRothblat, ILevitan. Modulation of endothelial inward-rectifier K+ current by optical isomers of cholesterol. Biophysical Journal, 2002, 83(6): 3211–3222
https://doi.org/10.1016/S0006-3495(02)75323-X
22 ILevitan, A E Christian, T N Tulenko, G H Rothblat. Membrane cholesterol content modulates activation of volume-regulated anion current in bovine endothelial cells. Journal of General Physiology, 2000, 115(4): 405–416
https://doi.org/10.1085/jgp.115.4.405
23 S LNiu, D C Mitchell, B J Litman. Manipulation of cholesterol levels in rod disk membranes by methyl-β-cyclodextrin effects on receptor activation. Journal of Biological Chemistry, 2002, 277(23): 20139–20145
https://doi.org/10.1074/jbc.M200594200
24 UKlein, G Gimpl, FFahrenholz. Alteration of the myometrial plasma membrane cholesterol content with β-cyclodextrin modulates the binding affinity of the oxytocin receptor. Biochemistry, 1995, 34(42): 13784–13793
https://doi.org/10.1021/bi00042a009
25 CRoduit, F G van der Goot, P De Los Rios, AYersin, PSteiner, GDietler, SCatsicas, FLafont, SKasas. Elastic membrane heterogeneity of living cells revealed by stiff nanoscale membrane domains. Biophysical Journal, 2008, 94(4): 1521–1532
https://doi.org/10.1529/biophysj.107.112862
26 A MBronder, A Bieker, SElter, MEtzkorn, DHäussinger, FOesterhelt. Oriented membrane protein reconstitution into tethered lipid membranes for AFM force spectroscopy. Biophysical Journal, 2016, 111(9): 1925–1934
https://doi.org/10.1016/j.bpj.2016.08.051
27 ICasuso, J Khao, MChami, PPaul-Gilloteaux, MHusain, J PDuneau, HStahlberg, J NSturgis, SScheuring. Characterization of the motion of membrane proteins using high-speed atomic force microscopy. Nature Nanotechnology, 2012, 7(8): 525–529
https://doi.org/10.1038/nnano.2012.109
28 J LHutter, J Bechhoefer. Calibration of atomic-force microscope tips. Review of Scientific Instruments, 1993, 64(7): 1868–1873
https://doi.org/10.1063/1.1143970
29 I NSneddon. The relation between load and penetration in the axisymmetric Boussinesq problem for a punch of arbitrary profile. International Journal of Engineering Science, 1965, 3(1): 47–57
https://doi.org/10.1016/0020-7225(65)90019-4
30 RMatzke, K Jacobson, MRadmacher. Direct, high-resolution measurement of furrow stiffening during division of adherent cells. Nature Cell Biology, 2001, 3(6): 607–610
https://doi.org/10.1038/35078583
31 AEmad, W F Heinz, M D Antonik, N P D’Costa, S Nageswaran, C ASchoenenberger, J HHoh. Relative microelastic mapping of living cells by atomic force microscopy. Biophysical Journal, 1998, 74(3): 1564–1578
https://doi.org/10.1016/S0006-3495(98)77868-3
32 R SLam, A R Shaw, M Duszyk. Membrane cholesterol content modulates activation of BK channels in colonic epithelia. Biochimica et Biophysica Acta (BBA)— Biomembranes, 2004, 1667(2): 241–248
https://doi.org/10.1016/j.bbamem.2004.11.004
33 MToselli, G Biella, VTaglietti, ECazzaniga, MParenti. Caveolin-1 expression and membrane cholesterol content modulate N-type calcium channel activity in NG108-15 cells. Biophysical Journal, 2005, 89(4): 2443–2457
https://doi.org/10.1529/biophysj.105.065623
34 HOh, E R Mohler III, A Tian, TBaumgart, S LDiamond. Membrane cholesterol is a biomechanical regulator of neutrophil adhesion. Arteriosclerosis, Thrombosis, and Vascular Biology, 2009, 29(9): 1290–1297
https://doi.org/10.1161/ATVBAHA.109.189571
35 SCorvera, C DiBonaventura, H SShpetner. Cell confluence-dependent remodeling of endothelial membranes mediated by cholesterol. Journal of Biological Chemistry, 2000, 275(40): 31414–31421
https://doi.org/10.1074/jbc.M001708200
36 DFrankel, J Pfeiffer, ZSurviladze, AJohnson, JOliver, BWilson, ABurns. Revealing the topography of cellular membrane domains by combined atomic force microscopy/fluorescence imaging. Biophysical Journal, 2006, 90(7): 2404–2413
https://doi.org/10.1529/biophysj.105.073692
37 JKwik, S Boyle, DFooksman, LMargolis, M PSheetz, MEdidin. Membrane cholesterol, lateral mobility, and the phosphatidylinositol 4,5-bisphosphate-dependent organization of cell actin. Proceedings of the National Academy of Sciences of the United States of America, 2003, 100(24): 13964–13969
https://doi.org/10.1073/pnas.2336102100
38 F JByfield, S Tikku, G HRothblat, K JGooch, ILevitan. OxLDL increases endothelial stiffness, force generation, and network formation. Journal of Lipid Research, 2006, 47(4): 715–723
https://doi.org/10.1194/jlr.M500439-JLR200
39 XZhang, J Hurng, D LRateri, ADaugherty, G WSchmid-Schönbein, H YShin. Membrane cholesterol modulates the fluid shear stress response of polymorphonuclear leukocytes via its effects on membrane fluidity. American Journal of Physiology. Cell Physiology, 2011, 301(2): C451–C460
https://doi.org/10.1152/ajpcell.00458.2010
40 TBorbiev, C Radel, VRizzo. Participation of caveolae in β1 integrin-mediated mechanotransduction. FASEB Journal, 2007, 21(6): A752–A752
41 YQi, L Andolfi, FFrattini, FMayer, MLazzarino, JHu. Membrane stiffening by STOML3 facilitates mechanosensation in sensory neurons. Nature Communications, 2015, 6(1): 8512
https://doi.org/10.1038/ncomms9512
42 F JByfield, H Aranda-Espinoza, V GRomanenko, G HRothblat, ILevitan. Cholesterol depletion increases membrane stiffness of aortic endothelial cells. Biophysical Journal, 2004, 87(5): 3336–3343
https://doi.org/10.1529/biophysj.104.040634
43 BHissa, B Pontes, P M SRoma, A PAlves, C DRocha, T MValverde, P H NAguiar, F PAlmeida, A JGuimaraes, CGuatimosim, et al.. Membrane cholesterol removal changes mechanical properties of cells and induces secretion of a specific pool of lysosomes. PLoS One, 2013, 8(12): e82988
https://doi.org/10.1371/journal.pone.0082988
44 R EBrown. Sphingolipid organization in biomembranes: What physical studies of model membranes reveal. Journal of Cell Science, 1998, 111(1): 1–9
45 APralle, P Keller, E LFlorin, KSimons, J HHörber. Sphingolipid-cholesterol rafts diffuse as small entities in the plasma membrane of mammalian cells. Journal of Cell Biology, 2000, 148(5): 997–1008
https://doi.org/10.1083/jcb.148.5.997
46 TWakatsuki, B Schwab, N CThompson, E LElson. Effects of cytochalasin D and latrunculin B on mechanical properties of cells. Journal of Cell Science, 2001, 114(5): 1025–1036
47 NKhatibzadeh, S Gupta, BFarrell, W EBrownell, BAnvari. Effects of cholesterol on nano-mechanical properties of the living cell plasma membrane. Soft Matter, 2012, 8(32): 8350–8360
https://doi.org/10.1039/c2sm25263e
48 LZhang, W F D Bennett, T Zheng, P KOuyang, X POuyang, X QQiu, A QLuo, MKarttunen, PChen. Effect of cholesterol on cellular uptake of cancer drugs pirarubicin and ellipticine. Journal of Physical Chemistry B, 2016, 120(12): 3148–3156
https://doi.org/10.1021/acs.jpcb.5b12337
49 ORamprasad, G Srinivas, K SRao, PJoshi, J PThiery, SDufour, GPande. Changes in cholesterol levels in the plasma membrane modulate cell signaling and regulate cell adhesion and migration on fibronectin. Cytoskeleton, 2007, 64(3): 199–216
https://doi.org/10.1002/cm.20176
50 C ALópez, A Hde Vries, S JMarrink. Molecular mechanism of cyclodextrin mediated cholesterol extraction. PLoS Computational Biology, 2011, 7(3): e1002020
https://doi.org/10.1371/journal.pcbi.1002020
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