Dynamic parameter of membrane lipid in lung cancer cell lines, carcinogenesis cells and cancer cells isolated from patients with lung cancer (original) (raw)
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Membrane fluidity characteristics of human lung cancer
Cancer Letters, 1999
Membrane¯uidity of non-cultured lung cancer tissue was studied by electron paramagnetic resonance (EPR). EPR spectra of a lipophilic spin probe in a tissue of resected tumor samples from 51 patients were compared with computer simulated spectra, which were superimpositions of spectra characterizing membrane domains with different¯uidity. The membranes of tumor tissues were more¯uid, than those of normal lungs; the most¯uid domains were enlarged and their order parameter decreased in comparison to normal tissue. An empirical¯uidity parameter (H13) was de®ned as the criterion to correlate EPR and clinical data. The histology of tumor, the quantitative presence of different tumor and non-tumor cells and the pathohisthological stage of the disease had no signi®cant in¯uence on¯uidity.
Applied Radiation and Isotopes, 1996
The changes in the microviscosity of the nuclear membranes of tumor and liver cells of tumor hosts with developing Erlich ascites carcinoma at different times after irradiation of lethal dose has been studied by spin probe method. Using two iminoxyl radicals localized in various lipid regions, it was shown that the character and degree of changes in microviscosity, estimated from rotational correlation time for spin probes, indicate the different response to irradiation of liver and tumor cells.
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.
Membrane fluidity affects tumor-cell motility, invasion and lung-colonizing potential
International Journal of Cancer, 1989
Membrane fluidity, determined by steady-state fluorescence polarization measurements, was correlated with metastatic capacity of murine tumor-cell lines. A correlation was observed in cell lines with different metastatic potential, and was confirmed when their lung-colonizing ability was modulated by alteration of either the membrane lipid composition or the culture conditions. Two cellular functions, motility and basement membrane invasion, were affected by the membrane lipid composition, and might explain the role of membrane fluidity observed in cancer metastasis.
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
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.
Lipid composition and lateral diffusion in plasma membranes of teratocarcinoma-derived cell lines
Cell, 1981
We measured lipid lateral diffusion rates for a series of teratocarcinoma-derived and embryo-derived cell lines, using the technique of fluorescence photobleaching recovery with a fluorescent lipid probe, CIGdil. The probe diffuses more rapidly in plasma membranes of embryonal carcinoma cells than in plasma membranes of teratocarcinoma-derived endodermal cell lines. When embryonal carcinoma cells are induced to differentiate by treatment with retinoic acid, diffusion constants of CIsdil are reduced to levels typical of endoderm. These changes are paralleled by differences in membrane cholesterol content; membrane free cholesterol levels in embyronal carcinoma lines are approximately half those found in endodermal lines, and are markedly increased upon retinoic-acid-induced differentiation.
Application of POLARIC™ fluorophores in an in vivo tumor model
Oncology Reports, 2013
Fluorescent and luminescent tools are commonly used to study the dynamics of cancer progression and metastases in real-time. Fluorophores have become essential tools to study biological events. However, few can sustain fluorescence long enough during long-term studies. In the present study, we focused on a series of new amphiphilic fluorophores known as POLARIC™, which emit strong fluorescence in lipid bilayers and can be readily modified using the Suzuki-Miyaura cross-coupling reaction. Appropriate chemical modifications of substituent groups can improve target-site specificity, reduce cytotoxicity and prolong emission. Therefore, in contrast to conventional fluorescent probes, these fluorophores show promise for long-term monitoring of biological processes. In the present study, we conducted long-term observations of tumor growth and metastasis using a POLARIC derivative as a novel fluorescent probe. For this purpose, we studied the metastatic melanoma cell line A375-SM, which proliferates at a high rate. We compared the characteristics of the POLARIC probe with the commercially available fluorescent dye PKH26 and fluorescent protein mRFP1. A375-SM cells were labeled with these fluorescent probes and orthotopically implanted into nude mice. The fluorescence emitted by POLARIC was detected more than five weeks after implantation without causing detectable harmful effects on tumor growth. By contrast, fluorescence of cells labeled with PKH26 could not be detected at this same time. Furthermore, POLARIC-, but not PKH26-labeled cells, were also detected in lung metastases. These results indicate that labeling cells with POLARIC fluorophores can significantly extend the time course of in vivo studies on tumor cell growth.
The effect of X-irradiation on membrane lipids of lymphosarcoma cells in vivo and in vitro
Journal of Radiation Research, 1982
Membrane lipids/Radiation/Tumor Phospholipids of the membranes of spleen lymphosarcoma cells were radioactively labelled with the radioresistant fatty acid palmitic acid (16:0) and the radiosensitive fatty acid arach idonic acid (20:4). The effect of irradiation on the specific radioactivity of the phospholipids was studied. For the in vivo experiments trace amounts of radioactive palmitate or arachidonate were injected intraperitoneally into lymphosarcoma-bearing mice. Incorporation of label into the lipids of the tumorous spleen cells was monitored in control animals and in mice that were whole body irradiated after injection of the label. In both groups of animals the label was detected in the blood within minutes after injection and was found to be efficiently incorpo rated into the phospholipids of the tumor cells. In the irradiated animals a marked transient increase in label incorporation was observed as compared to control animals. The radiation effect was observed in the lipids of the total homogenate, purified nuclei, the mitochondrial lysosomal fraction and in the microsomal fraction. Most experiments were performed with nuclei, which are known for their high radiosensitivity. The levels of label incorporation for palmitate and arachidonate were increased to the same extents and found to be dose depend ent. For a dose of 5 Gy the increased label incorporation started immediately after irradiation and lasted for a period of about 50 minutes. The increase in label incorporation into the phospholipids was preceded by an increase in the concentration of fatty acids in the cytosol of the tumors. Our experiments point to the occurrence of a transient increase in the flux of fatty acids through the plasma membrane as a result of irradiation and suggest that under normal physiological conditions fatty acid uptake through the plasma membrane is the rate-limiting step in the incorporation of acyl groups into the phospholipids. Experiments with isolated tumor cells also showed an increased incorporation of fatty acids into the phospholipids after irradiation. Again the incorporation patterns of both types of fatty acids (16:0) and (20:4) were very similar. A hypotonic treatment of the cells also resulted in a similar increase in fatty acid incorporation as irradiation did; the effects of hypo tonic treatment and irradiation were not additive.