Lipid composition and lateral diffusion in plasma membranes of teratocarcinoma-derived cell lines (original) (raw)
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Tumorigenicity of cell lines with altered lipid composition
Proceedings of the National Academy of Sciences, 1984
A series of closely related mouse fibroblast cell lines that differ in their content of neutral ether-linked glycerolipid and fatty acids has been used to investigate the relationship between lipid composition and tumorigenicity. Although these cell lines, derived from the same parental culture, were selected without reference to transformation or tumorigenicity, their ability to form tumors in irradiated mice was found to be closely correlated with ether-lipid content. The cell line with the highest level of ether-lipid (designated F40) produces more tumors, the tumors appear more rapidly than when parental cells are injected, and the number of F40 cells required for tumor induction is less by a factor of approximately equal to 1000. F40 tumors are highly invasive, readily metastasize, and rarely regress, in contrast to the occasional benign tumors produced by the parental cell line. Cell lines that are intermediate in their lipid composition appear to be intermediate in tumorigeni...
1991
hospta/, Be/flag With the aid of the Fluorescent lipophilic probe DPH tl. 6-diphenyl-1, 3, 5-hexatriene ), the degree of microvisccaiW OI) and lipid fluidity (LFU) obtained from lung cancer lines and carcinogenesis calls induced by irradiation as well as the patients with lung cancer were quantitatively monitored by Fluorescence polarization. The results have shown a marked decreased in q and a significant increase in LFU in various tumor cells as compared to normal calls. Sometime, the degree of fluidity in carcinogenesis cells induced by radiation and the patients with lung cancer have shown to be similar pattern. The possibility that these dynamic parmneter may serve as a diagnostic tool for an early detection of lung cancer is discussed.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1979
The fluorescence photobleaching recovery method has been used to determine the lateral mobilities of membrane lipids and proteins during the cell cycle of synchronized C1300 mouse neuroblastoma cells (clone Neuro-2A). As probes for lipid mobility, 3,3'-dioctadecylindocarbocyanine iodide and a fluorescein-labeled analog of ganglioside GM1 were used. Membrane proteins were labeled with rhodaminelabeled rabbit antibodies against mouse E14 cells. For both lipid probes the diffusion coefficients reach a minimum in mitosis, increase 2-to 3-fold during GI, remain constant at maximal values during S, and decrease again shortly before mitosis. Membrane proteins also exhibit minimum diffusion coefficients in mitosis, followed by a similar rise in GI. However, as cells proceed through S and G2, the lateral mobility of the membrane proteins gradually decreases. It is argued that lipid mobility is controlled by the fluidity of the membrane lipid matrix whereas protein mobility is governed also by other constraints.
MedChemComm, 2014
Dimethylsulfoxide (DMSO) for cell culture, Dulbecco's Modified Eagle Medium (DMEM), coumarin 153 (C153, Exciton, Scheme 1A) and 4′, 6-diamidino-2-phenylindole dihydrochloride (DAPI, Scheme1B) were purchased from Sigma Aldrich. Fetal bovine serum was purchased from Invitrogen. All the materials were used without further purification. Human lung cancer (A549) cell line were purchased from National Centre for Cell Science, Pune, India and cultured in our laboratory. Lung fibroblast cell was received as a gift from Dr. D. Sinha (IACS, Kolkata) and cultured in our laboratory. 2. Methods 2.1 Cell Preparation: Human lung cancer (A549) cells and non-cancer lung fibroblast (WI38) cells were grown in phenol red free DMEM with 10% fetal bovine serum, 1% Pen Strep Glutamine (from Gibco) in an atmosphere of 5% (v/v) CO 2 enriched air at 37 °C. A stock solution of C153 in biocompatible DMSO and DAPI in water (500 nM) were prepared. Cells were seeded at a density of 5000 cells per petri dish in a culture petri dish (BD BioCoat) for 18-24 hours before the dyes treatment. For proper staining of the cells, 200 µL of 500 nM dye solutions was added to the culture dish and incubated (half an hour for DAPI and 4 hours for C153) separately. After incubation the cells were washed 3-4 times with phosphate buffered
Lipidomic Analysis of Cancer Cell and Tumor Tissues
Methods in molecular biology, 2019
Due to their role in cellular structure, energetics, and signaling, characterization of changes in cellular and extracellular lipid composition is of key importance to understand cancer biology. In addition, several mass spectrometry-based profiling as well as imaging studies have indicated that lipid molecules may be useful to augment existing biochemical and histopathological methods for diagnosis, staging, and prognosis of cancer. Therefore, analysis of lipidomic changes associated with cancer cells and tumor tissues can be useful for both fundamental and translational studies. Here, we provide a high-throughput single-extractionbased method that can be used for simultaneous lipidomic and metabolomic analysis of cancer cells or healthy or tumor tissue samples. In this chapter, a modified Bligh-Dyer method is described for extraction of lipids followed by analysis of fatty acid composition by gas chromatography-mass spectrometry (GC-MS) or untargeted lipidomics using electrospray ionization mass spectrometry (ESIMS) coupled with reversephase (RP) ultraperformance liquid chromatography (UPLC) followed by multivariate data analysis to identify features of interest.
Lipid Composition, Physical State, and Lipid Peroxidation of Tumor Membranes
Toxicologic Pathology, 1984
Studies were carried out on microsomes isolated from the highly differentiated (slow-growing) Morris hepatoma 9618A, on microsomes and plasma membranes from the poorly differentiated (fast-growing) Morris hepatoma 3924A, and rat liver used as control. The lipid composition (phospholipid and cholesterol content, degree of fatty acid unsaturation) and peroxidation of such membranes has been correlated with the order and fluidity of the membrane bilayer. The results indicate that substrate availability is the rate-limiting step in microsomal and plasma membrane lipid peroxidation of hepatoma 3924A. From diphenylhexatriene fluorescence depolarization measurements it appears that the changes in lipid composition cause an increase in the order of the lipid bilayer on going from the control to hepatoma 9618A and 3924A microsomes, while fluidity is virtually unchanged. Conversely, for similar chemical changes, in plasma membranes from hepatoma 3924A the order is nearly the same and there is a decrease in fluidity. The changes in the above parameters of tumor membranes might be partly related to the loss of protective enzymes against oxygen radicals. This is supported by the observation that inhibition of liver superoxide dismutase and glutathione reductase, by treatment of rats with diethyldithiocarbamate and chloroethyl nitrosourea, respectively, renders the microsomal membranes more resistant to lipid peroxidation in vitro.
Lateral mobility of plasma membrane lipids in normal and transformed keratinocytes
Biochemical and Biophysical Research Communications, 1988
In this study we have examined possible differentiation-dependent modulations in plasma membrane lipid properties in normal keratinocytes, SV-40 transformed keratinocytes (SVKI4) and a number of squamous carcinoma (SCC) cells. In normal keratinocytes the lateral diffusion coefficient of plasma membrane lipids (D) differs sigllificant ly for cells cq~tured permanentl~ under low and normal Ca=~-conditions (5.16 x i0-> and 3.27 x i0-cm2/.s, respectively). When differentiation is induced by exposing low_~a2~-cultured cells to normal Ca 2~ concentrations D increases to 7.07 x i0 cm2/s during the initial hours of differentiation followed by a gradual sustained decrease to values also observed in cells cultured permanently under normal Ca2~-conditions. In SCC and SVKI4 cells a similar initial transient increase in lateral lipid mobility is observed upon initiation of differentiation, but, in contrast to normal keratinocytes, n~ sustained decrease in D is seen upon prolonged culturing under normal Ca ~° conditions. The results i/}dicate that the deficiency of the transformed cells to respond to Ca 2'-induced differentiation might involve transformation-dependent alterations in membrane structure and function. © z~88 Academlc Press, Inc. Cultured keratinocytes provide an attractive model to investigate differentiation-related modulations of cellular metabolism, since the extent of their capacity to differentiate can be readily manipulated by changing the extracellular Ca2+-concentration. The capacity to differentiate was monitored by the ability of the cells to form cornified envelopes.
Experimental Cell Research, 1998
lished [1, 2]. Many processes associated with cell The effect of various differentiation inducers on growth, differentiation, and cellular function are acmembrane cell dynamics was studied using HL-60 and companied by changes in membrane order parameter K562 leukemic cell lines. Membrane lipid dynamics was and fluidity [1]. These biophysical characteristics are measured by the steady-state fluorescence polarization dictated by changes in membrane-lipid-protein inter-(P) method utilizing either 1,6-diphenyl-1,3,5-hexaactions. Changes in membrane order parameters may triene (DPH) or the trimethyl ammonium derivative of be associated with the mechanism of signal transduc-DPH (TMA-DPH), which ascertains anchorage of the tion and/or may result in variable exposure of surface label to the membrane-water-lipid interface. Decrease receptors and antigens, thus modulating cellular funcin membrane microfluidity was observed in HL-60 cells tion [1, 2]. undergoing differentiation into macrophages by 1,25-Changes in lipid bilayer dynamics can be studied by dihydroxyvitamin D 3 and by K562 cells induced to difhydrophobic membrane probes such as 1,6-diphenylferentiate by DMSO. Sodium butyrate caused an in-1,3,5-hexatriene (DPH) and its trimethyl amino analog, crease in membrane fluidity in K562 cells undergoing 1-(4-trimethyl-ammonium phenyl)-6-phenyl-1,3,5-hexdifferentiation into erythroid-like cells while in HL-60 atriene, p-toluensulfonate (TMA-DPH). The measured cells a dual effect was observed. At 0.4 mM concentrachange in fluorescence anisotropy reveals alteration in tion, in which the cells were induced to differentiate the order parameter of the membrane lipid bilayer, ofalong the monocyte pathway, a decrease in membrane ten referred to as ''membrane microfluidity''; it refluidity was observed, while at 1 mM concentration an quires, however, the parallel measure of the fluoresincrease in membrane fluidity occurred. Interferon-g cence lifetime which is essential to ascertain that (IFN-g) induced an increase in membrane fluidity in changes in rotational correlation time cause the obboth cell lines. Using HL-60 cells fluorescently labeled by TMA-DPH, similar results indicating fluidization of served change in the measured anisotropy [1]. the membrane following IFN-g treatment were ob-The levels of protein and lipid constituents of plasma tained. Advanced fluorescence lifetime measurements, membranes show alterations during cell proliferation evaluated either by phase modulation spectrofluoromand differentiation. These changes can affect the physetry or by single photon correlation fluorometry conical state of the membrane. In early as well as recent firmed that the decrease in fluorescence polarization reports, higher fluidity has been found in immature by IFN-g resulted from membrane fluidization and not cells while lower fluidity has been documented in diffrom elongation of the probe's excited state lifetime. It ferentiated cells. Several cells, such as erythrocytes, is suggested that the inducer mode of action, and not lymphocytes, neutrophils, and liver cells, did show the differentiation route, determine the outcome of these alterations in fluidity [3-10]. Cyclic changes in changes in membrane microviscosity. ᭧ 1998 Academic Press membrane microviscosity during cell cycle have been reported [11]. Differentiated cells lose their proliferating capabilities. Thus, since it was shown that prolifer-1 To whom correspondence and reprint requests should be ad-The sensitivity of leukemia cell lines to various difdressed at The Chemistry Department, Ben-Gurion University of the ferentiation agents and their ability to undergo multi-Negev, Beer-Sheva 84 105, Israel.
European Journal of Biochemistry, 1992
The chemical composition of highly purified plasma membrane preparations from a series of malignant Chinese hamster ovary (CHO) cell lines were undertaken to ascertain if neutral lipid, including cholesteryl ester and triacylglycerol, were present. Triacylglycerols (33 -41 nmol/mg total lipid) and cholesteryl ester (226 -271 nmol/mg) were measured in the plasma membranes and differences in the chemical composition of these membranes recorded. The most significant difference was a gradual decrease in the level of free cholesterol from wild type (312 f 7 nmol/mg total plasma membrane lipid), Pod RII-6 (268 f 64 nmol/mg total plasma membrane lipid), Col R-22 (243 f 39 nmol/mg total plasma membrane lipid) to EOT (204 20 nmol/mg total plasma membrane lipid), with a concomitant increase in the degree of saturation of the cholesteryl ester fatty acids, particularly palmitic acid. No statistically significant differences were apparent in the chemical composition of the whole cells in this series.
Plasma membrane reorganization induced by tumor promoters in an epithelial cell line
Proceedings of the National Academy of Sciences, 1984
The effects of phorbol ester tumor promoters on the lateral diffusion in plasma membrane lipid environments were examined by the technique of fluorescence recovery after photobleaching. To this end, the probe coliarein, a fluorescent lipid analog that has the property of exclusive localization in the plasma membrane, was synthesized. Measured decreases in three parameters [percentage of fluorescence bleached (30%), percentage of recovery (52%), and half-time for recovery (52%)] connoted the appearance of an immobile fraction upon exposure to tumor promoters. These data are consistent with lipid reorganization in response to a reorganization of the intraand perimembranous macromolecular scaffolding upon the interaction of cells with tumor promoters. The idea of induced reorganization is supported by experiments in which cell shape change, brought about by either exposure to cytochalasin B or growth on matrices of collagen, fibronectin, or laminin, resulted in values in the fluorescence recovery after photobleaching technique similar to those with active phorbol esters. Membranes are dynamic structures in which proteins, glyco