| For medical practitioners and the general public - Cholesterol Journal Article Catalog. |
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| Trans fat. Does margarine really lower cholesterol? Beardsley, T. (1991), Sci Am 264(1): 34. |
| Trans fatty acid derived phospholipids show increased membrane cholesterol and reduced receptor activation as compared to their cis analogs Niu, S. L., D. C. Mitchell, et al. (2005), Biochemistry 44(11): 4458-65. Abstract: The consumption of trans fatty acid (TFA) is linked to the elevation of LDL cholesterol and is considered to be a major health risk factor for coronary heart disease. Despite several decades of extensive research on this subject, the underlying mechanism of how TFA modulates serum cholesterol levels remains elusive. In this study, we examined the molecular interaction of TFA-derived phospholipid with cholesterol and the membrane receptor rhodopsin in model membranes. Rhodopsin is a prototypical member of the G-protein coupled receptor family. It has a well-characterized structure and function and serves as a model membrane receptor in this study. Phospholipid-cholesterol affinity was quantified by measuring cholesterol partition coefficients. Phospholipid-receptor interactions were probed by measuring the level of rhodopsin activation. Our study shows that phospholipid derived from TFA had a higher membrane cholesterol affinity than their cis analogues. TFA phospholipid membranes also exhibited a higher acyl chain packing order, which was indicated by the lower acyl chain packing free volume as determined by DPH fluorescence and the higher transition temperature for rhodopsin thermal denaturation. The level of rhodopsin activation was diminished in TFA phospholipids. Since membrane cholesterol level and membrane receptors are involved in the regulation of cholesterol homeostasis, the combination of higher cholesterol content and reduced receptor activation associated with the presence of TFA-phospholipid could be factors contributing to the elevation of LDL cholesterol. |
| Trans fatty acids and cholesterol metabolism: mechanistic studies in rats and rabbits fed semipurified diets Gatto, L. M., M. A. Lyons, et al. (2001), Int J Food Sci Nutr 52(5): 435-41. Abstract: Studies were conducted in rabbits and rats to investigate the effects of diets rich in oleic (CIS diet), palmitic (SAT diet) and trans fatty acids (TRANS diet) on plasma lipids and lipoprotein metabolism. An important difference between these species is that rabbits possess plasma cholesteryl ester transfer protein (CETP) activity while rats are devoid of transfer activity. In the presence of dietary cholesterol (0.2% w/w) the change in plasma low density lipoprotein-cholesterol (LDL-C) concentration from baseline was significantly higher in rabbits fed the TRANS diet compared with those fed the CIS diet (P < 0.01). Despite this difference, the hepatic LDL-receptor activity was similar in all groups. Also, the fatty acid composition of hepatic phospholipids was affected by diet with lower proportion of palmitic (11%) and higher (19%) linoleic acid despite a similar content in the diet. These effects may represent the maintenance of membrane fluidity within narrow limits to ensure optimal function. The studies in rats showed that the plasma total cholesterol concentration was 20% lower (P < 0.01) in TRANS-fed rats compared with those fed the CIS diet. The results of an in vivo assay of reverse cholesterol transport (RCT) suggested that the three diets gave rise to high density lipoprotein (HDL) particles with similar capacity to accept cellular cholesterol. The differential effects of dietary trans fatty acids in these animal models provide another line of evidence that reinforces the significant role of CETP activity in determining the distribution of plasma cholesterol in response to dietary trans fatty acids. |
| Trans fatty acids, HDL-cholesterol, and cardiovascular disease. Effects of dietary changes on vascular reactivity de Roos, N. M., E. G. Schouten, et al. (2003), Eur J Med Res 8(8): 355-7. Abstract: A high consumption of trans fatty acids increases the risk of cardiovascular disease (CVD). We investigated whether this increase in risk was due to the decrease in serum HDL-cholesterol by trans fatty acids, because low concentrations of serum HDL-cholesterol also increase risk of CVD. Flow-mediated vasodilation (FMD) was used as an endpoint in dietary interventions that were designed to change the concentration of serum HDL-cholesterol within 4 weeks in healthy volunteers. Replacement of 10% of energy from saturated by trans fatty acids decreased serum HDL-cholesterol by 21 % and impaired FMD. However, a replacement of monounsaturated fats by carbohydrates did not impair FMD, although it decreased serum HDL-cholesterol by 13%. Acute postprandial impairments of FMD by either trans fats or saturated fats were not found, suggesting that long-term effects are responsible for the detrimental effect of trans fats on health. However, the role of serum HDL-cholesterol appears to be less than we expected. |
| Trans monounsaturated fatty acids and serum cholesterol levels Grundy, S. M. (1990), N Engl J Med 323(7): 480-1. |
| Transbilayer complementarity of phospholipids in cholesterol-rich membranes Zhang, J., B. Jing, et al. (2005), Biochemistry 44(9): 3598-603. Abstract: Lipid-lipid interactions across cholesterol-rich phospholipid bilayers were investigated by measuring nearest-neighbor preferences of exchangeable phospholipids derived from 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), in the presence of nonexchangeable dimers (i.e., templates) made from DMPE or DSPE. When homotemplates were present, a significant preference for homophospholipid association was observed. In contrast, when the corresponding heterotemplate was present, heterodimer formation was favored. These results support a model in which the longer phospholipid in one monolayer preferentially associates with the shorter one in the adjoining monolayer. In the absence of cholesterol, transbilayer complementarity was also observed but to a lesser degree. Transbilayer complementarity of phospholipids is likely to play an important role in stabilizing biological membranes and in promoting a compositional interdependence of their two lipid leaflets. |
| Transbilayer distribution of cholesterol is modified in brain synaptic plasma membranes of knockout mice deficient in the low-density lipoprotein receptor, apolipoprotein E, or both proteins Igbavboa, U., N. A. Avdulov, et al. (1997), J Neurochem 69(4): 1661-7. Abstract: Both apolipoprotein E (apoE) and the low-density lipoprotein (LDL) receptor are present in brain; however, little is known regarding the function of these proteins in brain, in particular with respect to brain cholesterol. The role of apoE and the LDL receptor in modulating the transbilayer or asymmetric distribution of cholesterol in the exofacial and cytofacial leaflets of synaptic plasma membranes (SPMs) was examined in mutant mice deficient in apoE, the LDL receptor, or both proteins by using the fluorescent sterol dehydroergosterol and fluorescent quenching procedures. Fluidity of the exofacial and cytofacial leaflets was also measured. Cholesterol asymmetry of SPMs was altered in the mutant mice, with the largest effect observed in the LDL receptor-deficient mice. There was an approximately twofold increase in the percent distribution of cholesterol in the exofacial leaflet of the LDL receptor-deficient mice (32%) compared with C57BL/6J mice (15%). Mice deficient in apoE or both proteins also showed a significantly higher percent distribution of cholesterol (23 and 26%, respectively) in the exofacial leaflet compared with the C57BL/6J mice. Although the percent distribution of cholesterol was highest in the exofacial leaflet of the LDL receptor-deficient mice, fluidity of the exofacial leaflet of that group was significantly lower. However, the cholesterol-to-phospholipid ratio of SPMs of the LDL receptor-deficient mice was significantly lower, and this difference was largely the result of a significant increase in the total amount of SPM phospholipid. This study demonstrates for the first time that SPM lipid structure is altered in mice deficient in apoE or the LDL receptor. Although the mechanism that maintains the asymmetric distribution of cholesterol in plasma membranes is not well understood, data of the present experiments indicate that both apoE and the LDL receptor are involved in maintaining the transbilayer distribution of cholesterol. |
| Transbilayer movement and net flux of cholesterol and cholesterol sulfate between liposomal membranes Rodrigueza, W. V., J. J. Wheeler, et al. (1995), Biochemistry 34(18): 6208-17. Abstract: The kinetics of cholesterol and cholesterol sulfate (CS) movement between vesicles have been investigated. CS is widely distributed in cell membranes, plasma and skin and is similar in structure to cholesterol, but possesses an ionizable sulfate moiety at the 3 beta-position which imparts a negative charge at physiological pHs. Donor vesicles of various sizes ranging from 40 to 250 nm, composed of egg phosphatidylcholine (EPC)/sterol/N-palmitoyldihydrolactosylcerebroside (75:10:15 mole ratio) containing trace amounts of 3Hsterol, were used to monitor sterol transfer into a 10-fold excess of large unilamellar vesicles (LUV) composed of EPC with a diameter of 100 nm. The two populations of vesicles were separated by centrifugation following the addition of a lectin which caused the aggregation of donor vesicles. Both cholesterol and CS exhibited biphasic kinetics of exchange. The rate constants for efflux and transbilayer diffusion for both sterol molecules were determined after fitting kinetic data, using numerical integration, to a three-compartment model, which includes the inner and outer monolayers of donor vesicles and the acceptor bilayer. The rate of intermembrane exchange for CS was approximately 10-fold faster than for cholesterol in all liposomes tested. Using the kinetic model, a rate of transbilayer movement for cholesterol and CS was estimated. In both cases, it was found to be slower than the rate of efflux from the surface of vesicles. For vesicles containing CS, the surface charge was monitored to demonstrate that the slowly exchanging pool was located in the inner monolayer, and the rapidly exchanging pool in the outer half of the bilayer. For cholesterol, it was not possible to distinguish between this model and one where lateral domains of cholesterol within the plane of the bilayer may influence the kinetics of exchange. |
| Trans-bilayer movement of erythrocyte membrane cholesterol in human essential hypertension Frithz, G. (1995), J Hypertens 13(12 Pt 1): 1480-81. |
| Transbilayer movement of erythrocyte membrane cholesterol in human essential hypertension Muriana, F. J., C. Montilla, et al. (1995), J Hypertens 13(6): 619-23. Abstract: OBJECTIVE: To study whether the rate of transbilayer movement of membrane cholesterol is impaired in erythrocyte membrane of normo- and hypercholesterolaemic patients with untreated essential hypertension. DESIGN: An observational case-control study. METHODS: Erythrocytes were prepared from venous blood samples obtained from normotensive subjects and hypertensive patients. The rate of transbilayer movement of membrane cholesterol was monitored in intact erythrocytes, using a radiolabelled cholesterol tracer. Erythrocytes were treated briefly or continuously with cholesterol oxidase to convert a portion of the outer leaflet cholesterol to cholestenone, and the specific radioactivity of cholestenone was determined over the period of tracer equilibration. The decrease in specific radioactivity of cholestenone reflected the transbilayer movement of radiolabelled cholesterol. RESULTS: There were no significant differences between the diffusion of cholesterol across the erythrocyte membrane of normo- and hypercholesterolaemic hypertensive patients, but the rates were significantly lower than that estimated in control subjects. The mean +/- SD half-times for the process were 55.1 +/- 8.8 and 11.3 +/- 2.1 min in controls, 63.1 +/- 9.2 and 15.8 +/- 2.3 min in normocholesterolaemic hypertensive patients, and 66.2 +/- 9.4 and 16.2 +/- 1.7 min in hypercholesterolaemic hypertensive patients, after a brief and after a continuous cholesterol oxidase treatment, respectively. CONCLUSION: There is a reduction in the transbilayer movement of membrane cholesterol in erythrocytes of patients with untreated essential hypertension. |
| Transbilayer movement of fully ionized taurine-conjugated bile salts depends upon bile salt concentration, hydrophobicity, and membrane cholesterol content Donovan, J. M. and A. A. Jackson (1997), Biochemistry 36(38): 11444-51. Abstract: Taurine-conjugated bile salts mediate rapid transmembrane flux of divalent cations, irrespective of whether bile salts and divalent cations are initially on the same or opposite side of the membrane. We therefore hypothesized that ionized bile salts can equilibrate between membrane hemileaflets. We quantitated bile salt binding to large unilamellar egg yolk phosphatidylcholine (EYPC) +/- cholesterol (Ch) vesicles under conditions in which one or both hemileaflets were initially exposed to bile salts. At unbound taurodeoxycholate (TDC) concentrations >0.2 mM, the dependence of binding on TDC concentration after 30 min was indistinguishable for vesicles prepared by either method and did not change from 30 minutes to 24 h. At unbound TDC concentrations <0.1 mM, the ratio of bound/free TDC to EYPC vesicles doubled over a single exponential time course. Equilibration times were greater for the more hydrophilic bile salts taurocholate and tauroursodeoxycholate, for EYPC/Ch vesicles, and at lower temperatures. For glycine-conjugated bile salts, time-dependent changes in binding did not occur, consistent with more rapid equilibration of the small fraction of the protonated form. We conclude that fully ionized conjugated bile salts translocate between lipid bilayer hemileaflets, in contrast to previous observations that equilibration of fully ionized unconjugated bile salts occurs at a negligible rate in small unilamellar vesicles. The rate of "flip-flop" increases with increases in intramembrane bile salt concentration and hydrophobicity but decreases with cholesterol content and lower temperature. We speculate that physiologically, even in the absence of a specific membrane transporter, bile salts can gain access to intracellular compartments and mediate increases in divalent cation flux that may underlie cytotoxicity. |
| Transcriptional control mechanisms in the regulation of cholesterol balance Osborne, T. F. (1995), Crit Rev Eukaryot Gene Expr 5(3-4): 317-35. Abstract: The mechanisms that govern regulation of cholesterol metabolism in higher eukaryotic cells provide an example of how metabolic regulation has evolved to establish growth and nutritional control in a multicellular environment. Two sources of cholesterol must be balanced to ensure optimum growth and viability. Much of the control is established by regulating the levels of key proteins involved in cholesterol uptake and biosynthesis and this occurs by alterations in promoter activity. The studies discussed here track the progression in understanding the mechanism for transcriptional regulation by cholesterol from the isolation of the key genes involved, to the careful dissection of the cis-acting sequences that control expression, and on to what is currently known about the trans-acting proteins that mediate the regulatory response. |
| Transcytotic efflux from early endosomes is dependent on cholesterol and glycosphingolipids in polarized hepatic cells Nyasae, L. K., A. L. Hubbard, et al. (2003), Mol Biol Cell 14(7): 2689-705. Abstract: We examined the role that lipid rafts play in regulating apical protein trafficking in polarized hepatic cells. Rafts are postulated to form in the trans-Golgi network where they recruit newly synthesized apical residents and mediate their direct transport to the apical plasma membrane. In hepatocytes, single transmembrane and glycolipid-anchored apical proteins take the "indirect" route. They are transported from the trans-Golgi to the basolateral plasma membrane where they are endocytosed and transcytosed to the apical surface. Do rafts sort hepatic apical proteins along this circuitous pathway? We took two approaches to answer this question. First, we determined the detergent solubility of selected apical proteins and where in the biosynthetic pathway insolubility was acquired. Second, we used pharmacological agents to deplete raft components and assessed their effects on basolateral-to-apical transcytosis. We found that cholesterol and glycosphingolipids are required for delivery from basolateral early endosomes to the subapical compartment. In contrast, fluid phase uptake and clathrin-mediated internalization of recycling receptors were only mildly impaired. Apical protein solubility did not correlate with raft depletion or impaired transcytosis, suggesting other factors contribute to apical protein insolubility. Examination of apical proteins in Fao cells also revealed that raft-dependent sorting does not require the polarized cell context. |
| Transdermal estrogen reduces vascular resistance and serum cholesterol in postmenopausal women West, S. G., A. L. Hinderliter, et al. (2001), Am J Obstet Gynecol 184(5): 926-33. Abstract: OBJECTIVE: Our aim was to compare the effects of transdermal versus oral estrogens on vascular resistance index, mean arterial pressure, serum lipid concentrations, norepinephrine, and left ventricular structure. STUDY DESIGN: Ten postmenopausal women received transdermal estradiol (0.05 mg/d) plus cyclic oral progesterone for 6 months. Responses were compared with those of 23 women receiving oral conjugated estrogens (0.625 mg/d) plus cyclic progesterone and with those of 9 subjects receiving placebo in a concurrent randomized trial. We assessed the vascular resistance index and the mean arterial pressure at rest and during behavioral stressors. RESULTS: Oral and transdermal estrogen significantly decreased the vascular resistance index, mean arterial pressure, norepinephrine, and total and low-density lipoprotein cholesterol to a similar extent. Changes in the vascular resistance index and mean arterial pressure were equally evident at rest and during stress. Although both treatments reduced left ventricular mass (-4% to -6%) and relative wall thickness (-3% to -5%), these changes were not statistically significant. CONCLUSIONS: Equivalent reductions in vascular resistance index, norepinephrine, mean arterial pressure, and cholesterol were observed with transdermal and oral estrogens. Future studies comparing novel hormone regimens with oral hormone replacement therapy should include multiple risk markers to allow better assessment of their potential impact on coronary artery health. |
| Transdermal estrogens do not appear to modify the extent of lesional areas of aortic atherosclerosis in oophorectomized rabbits on a cholesterol-rich diet Blumel, J. E., C. Castelo-Branco, et al. (2000), Atherosclerosis 148(2): 303-8. Abstract: Cardiovascular disease (CVD) is the leading cause of death in older women in industrialised countries. It has been suggested that it is the cessation of estrogen production by the ovaries that puts postmenopausal women at increased risk of CVD. Estrogen therapy has demonstrated a protective effect against CVD and several reports suggest that diverse mechanisms may be involved. Oral estrogen appears to be associated with a better lipid profile than the use of transdermal estrogens; however, it is assumed that estrogens, oral and non-oral, have direct actions on the blood vessels that may exert an important role in cardiovascular disease prevention. To investigate the effect of transdermal estrogen therapy on aorta atherogenesis, we studied 20 cholesterol-fed New Zealand White rabbits for 4 months. The rabbits were oophorectomized and randomly assigned to two groups. Ten rabbits underwent bilateral ovariectomy followed by treatment with transdermal 17-beta-estradiol (group E) and the other 10 received placebo after sterilization (Group C). After diet total levels of cholesterol increase in group C from 50. 0+/-12.5 to 820.9+/-186.0 mg/dl, and in group E from 52.6+/-9.4 to 811.4+/-213.0 mg/dl (no significant differences were observed between groups). Estrogen therapy increased twofold the total reactive antioxidant potential (TRAP group C: 22.5+/-16.7 mmol of Trolox/l vs. TRAP group E: 43.4+/-22.4 mmol of Trolox/l; P<0.04). At 4 months, the cholesterol-rich diet caused atherosclerotic lesions in both treated and untreated rabbits affecting 18.7+/-14.5 and 21. 6+/-9.7% of the aortic surface respectively. In summary, the principal result from this study was that although treatment with transdermal 17-beta-estradiol in cholesterol-fed ovariectomized rabbits increases the TRAP to pre-surgery values, it does not inhibit aortic cholesterol accumulation. |
| Transdifferentiation of mouse aortic smooth muscle cells to a macrophage-like state after cholesterol loading Rong, J. X., M. Shapiro, et al. (2003), Proc Natl Acad Sci U S A 100(23): 13531-6. Abstract: Mouse aortic smooth muscle cells (SMCs) were loaded for 72 h with cholesterol by using cholesterol:methyl-beta-cyclodextrin complexes, leading to approximately 2-fold and approximately 10-fold increases in the contents of total cholesterol and cholesteryl ester, respectively. Foam-cell formation was demonstrated by accumulation of intracellular, Oil Red O-stained lipid droplets. Immunostaining showed decreased protein levels of smooth muscle alpha-actin and alpha-tropomyosin and increased levels of macrophage markers CD68 and Mac-2 antigen. Quantitative real-time RT-PCR revealed that after cholesterol loading, the expression of SMC-related genes alpha-actin, alpha-tropomyosin, myosin heavy chain, and calponin H1 decreased (to 11.5 +/- 0.5%, 29.3 +/- 1.4%, 23.8 +/- 1.4%, and 3.8 +/- 0.5% of unloaded cells, respectively; P < 0.05 for all), whereas expression of macrophage-related genes CD68, Mac-2, and ABCA1 mRNA increased (to 709 +/- 84%, 330 +/- 11%, and 207 +/- 13% of unloaded cells, respectively; P < 0.05 for all), thereby demonstrating that the protein changes were regulated at the mRNA level. Furthermore, these changes were accompanied by a gain in macrophage-like function as assessed by phagocytotic activity. Expression of vascular cell adhesion molecule 1 and monocyte chemoattractant protein 1, known responders to inflammation, were not changed. In conclusion, cholesterol loading of SMC causes phenotypic changes regulated at the mRNA level that result in a transdifferentiation to a macrophage-like state. This finding suggests that not all foam cells in lesions may have a macrophage origin, despite what is indicated by immunostaining for macrophage-related markers. Furthermore, inflammatory changes in foam cells observed in vivo may not be simple consequences of cholesterol accumulation. |
| Transfection of myoblasts in primary culture with isomeric cationic cholesterol derivatives Bischoff, R., Y. Cordier, et al. (1997), Anal Biochem 254(1): 69-81. Abstract: Transfection of satellite cells from dog muscle (myoblasts) in primary culture has been optimized with respect to the position of the cholesteryl moiety along the polyamine chain of spermidine or spermine. Spermidine or spermine were derivatized with cholesterylchloroformate giving rise to three isomers in the case of spermidine and two isomers for spermine that were separated by reversed-phase high-performance liquid chromatography (rp-HPLC). The position of the cholesteryl moiety was assigned by 13C-NMR and coelution with synthetic isomers of defined structure. The isomeric cationic lipids were evaluated for their transfection activity in myoblasts from dog muscle and a human lung epithelial cell line (A549) using plasmid DNA expressing the luciferase reporter gene. The results showed that the position of the cholesteryl moiety is of critical importance for efficient transfection of myoblasts in primary culture with isomers having a derivatized secondary amine being significantly more effective than those with a derivatized primary amine. On the contrary, differences in the A549 cell line were less pronounced and did not follow the same pattern. The results show that slight structural differences between cationic lipids lead to significantly different transfection efficiencies for myoblasts in primary culture. This may also represent an advantage in view of cell or organ targeting. |
| Transfection of urothelial cells using methyl-beta-cyclodextrin solubilized cholesterol and Dotap Lawrencia, C., R. Mahendran, et al. (2001), Gene Ther 8(10): 760-8. Abstract: the murine urothelial cell line, mb49 was transfected with the reporter gene pcmvlacz using a number of commercial transfection agents. the transfection efficiency of these agents, as determined by beta-galactosidase activity, is in the order of dotap>superfect>Fugene. The addition of methyl-beta-cyclodextrin solubilized cholesterol (MBC) to Dotap and Superfect further improved their transfection efficiency by 3.8-fold and 2.6-fold, respectively. beta-Galactosidase activity was detectable within 1 h of transfection and peaked at 48 h. Nuclear and cytoplasmic separation showed that with Dotap + methyl-beta-cyclodextrin solubilized cholesterol (DMBC), the DNA plasmid complex was found in both the nucleus and the cytoplasm. In vivo, murine bladders were transfected with an intravesical instillation of DMBC + DNA for 2 h. Two days later the bladder, lungs, liver, spleen and heart were assayed for the presence of the beta-galactosidase gene by staining and PCR. Expression of the gene was confined to the bladder. Both in vitro and in vivo expression was observed after as little as a 15 min exposure to DMBC:DNA. Expression of the marker gene was present up to 30 days after transfection in vivo. From our data it appears that DMBC is the best nonviral agent for the transfection of urothelial cells in vitro and in vivo. |
| Transfer and expression of a Streptomyces cholesterol oxidase gene in Streptococcus thermophilus Somkuti, G. A., D. K. Solaiman, et al. (1991), Biotechnol Appl Biochem 13(2): 238-45. Abstract: The recombinant plasmid pNCO937 (8.1 kbp) containing a Streptomyces sp. cholesterol oxidase gene was introduced into Streptococcus thermophilus by electrotransformation. Transformation frequency was 7.2 x 10(5) colony forming units/micrograms of DNA. The presence of the cholesterol oxidase gene in S. thermophilus was confirmed with Southern blot analysis using a biotinylated probe. Thin-layer chromatographic analysis showed the expression of the Streptomyces cholesterol oxidase gene resulting in the oxidation of cholesterol to 4-cholesten-3-one. S. thermophilus may be a suitable host for the expression of other genes regulating prokaryotic cholesterol metabolism. |
| Transfer into a mesothelioma cell line of tumor suppressor gene p16 by cholesterol-based cationic lipids Piperno-Neumann, S., O. Oudar, et al. (2003), Biochim Biophys Acta 1611(1-2): 131-9. Abstract: In this work, the tumor suppressor gene p16 was efficiently transferred into FR cells isolated from a patient with malignant mesothelioma using cationic liposomes prepared from trimethyl aminoethane carbamoyl cholesterol (TMAEC-Chol) and triethyl aminopropane carbamoyl cholesterol (TEAPC-Chol). This transfer was performed after preliminary assays were undertaken to find the optimal transfection conditions. Results showed that an efficient transfer of plasmids containing the reporter gene pCMV-beta galactosidase vectorized by TMAEC-Chol/DOPE and TEAPC-Chol/DOPE liposomes into mesothelioma FR cells was obtained as assessed by luminometric measurements of beta-galactosidase activity. Cytotoxicity studied by MTT test showed that at concentrations used for this study, the cationic liposomes have no effect on cell growth. Transfer into mesothelioma FR cells of a plasmid construct containing the tumor suppressor gene p16 was carried out with these liposomes. Western blotting and immunofluorescence showed the presence of p16 in treated cells. An inhibition of cell growth was observed, indicating that efficient tumor suppressor gene transfer can be performed by using cationic liposomes. |