David S. Newburg, Ph.D.

EFFECT OF PHOSPHOGLYCOLIPID EXTRACT (NT FACTOR)

ON NORMAL AND CANCEROUS CELLS

Cancerous cells of tumors import phospholipids from normal cells.

One of the fundamental biochemical differences between tumor cells and normal cells is the composition of the membrane lipid, including glycosphingolipid and phospholipids.  The phospholipid content of tumor cell membranes in known to be distinct from that of normal cells (Bergelson 1974, Spangler 1975, Hostetler 1976, Burlakova 1980, 1991).  The difference in phospholipid content has been attributed to differences in rates of phospholipid transfer to the plasma membrane of aggressive tumors.  There are two components to this difference in the ability of tumor membranes to acquire phospholipids from neighboring cells:  first is a difference in phospholipid exchange due to phospholipid exchange protein (PLEP); second is a difference in phospholipid exchange rates due to the intrinsic lipid composition of the membrane.  Over 90% of the Phosphatidylcholine in hepatoma microsomes can be exchanged within two hours at 37⁰C (Palmina 1995).  In Morris hepatoma cells, the transfer activity of phosphatidylcholine was 2 to 3 times higher than in controls (Poorthuis 1980).  However, only some of this difference could be accounted for by an increase in PLEP activity, the rest being attributed to intrinsic differences in membrane lipids.  Clearly, many types of tumors are able to incorporate extrinsic phospholipids into their membranes at the expense of normal cells of the body with the potential to deplete the phospholipids in the normal cells.

Reduced levels of phospholipids in normal cells can limit metabolic activity and limit available energy.

Phospholipids, as part of the membrane structure, maintain membrane integrity, and, through changes in membrane fluidity, also regulate enzyme activities and membrane transport processes (Spector 1981, 1985).  Phospholipids can have other specific functions.  Signal transduction utilizes phospatidylcholine and phosphatidylinositol for the production of diacyl glycerol (DAG) by phospholipase C (Berridge 1989) and for the production of inositol triphosphate (IP₃) (Ranan 1990, Michell 1988, Margolis 1990).  One of the choline phospholipids (1-alkyl-2acetyl-SN-glycerol-3-phosphocholine) is the substrate for the synthesis of platelet activating factor (Synder 1989).  The arachidonic acid found as part of the structure of choline or inositol phospholipid is utilized for the production of prostaglandin and leukotriene (Nordoy 1990).  The choline of phosphatidylcholine may be used in neural tissue for the synthesis of acetylcholine (Blusztain 1987).

 Plasma brain and neuronal choline concentrations were elevated by oral administration of choline, which also causes the release of acetylcholine in the neuromuscular system (Haubrich 1976, Cohen 1976).  Furthermore, muscle function has been shown to decrease during choline deficiency (Zeisel 1990).  Physical stress depresses plasma choline concentration, e.g., individuals in the Boston Marathon of 1986 showed 40% decreases in plasma choline levels during the race (Conlay 1986).  Providing phosphatidylcholine prior to exercise can compensate for these choline losses (von Allwörden1993).  Even with shorter and less strenuous forms of exercise, a supplemental supply of lecithin results in an increase in performace (von Allwörden 1995).

When tumor cells sequester large amounts of the phosphatidylcholine produced by normal cells, this could lead to a loss of choline homeostasis, producing decreases in plasma, brain, and muscle choline that would be expected to result in muscle fatigue.  This could account for some of the malaise and chronic fatigue that is known to accompany certain forms of cancer.  Under these circumstances, exogenous oral supplements would be expected to provide some measure of relief from cancer-associated fatigue. 

The rate of phospholipid accumulation in cancer cells is independent of exogenous supply.

In general, adult tissues contain more phosphatidylcholine than immature tissues (Sun 1985, Yorek 1993).  Like immature developing tissues, some tumors contain lower levels of phospholipid than corresponding normal tissue (Bergelson 1975).  However the phospholipid content varies greatly from tumor to tumor.  Many varieties of cancerous tissue contain more phosphatidylcholine with increased amounts circulating in the blood and available for use by the tumor (Takenaka 1983, Nikolasev 1972, Aso 1981).  Thus, some tumors can deplete normal tissue of phospholipid.

NT Factor^TM phosphoglycolipid improves cell maintenance and metabolic activity of normal cells.

The integrity of mitochondria and their ability to produce energy can be measured by isolating lymphocytes, treating them with Rhodamine 123 (a mitochondrial stain), and analyzing them using FACSCAN, a flow cytometer modified for analysis of mitochondria.  In rats, there is a measurable decrease in mitochondrial function as the rat ages.  However, in rats fed a diet that contains NT Factor^TM phosphoglycolipid, mitochondria showed a 20% improvement over those fed the identical diet without the NT Factor^TM, as measured by Rhodamine flow cytometry.  (Michael Seidman, personal communication)

Assuming that the degradation of mitochondrial function with age is caused by cumulative chemical toxicity, it would appear that NT Factor^TM phosphoglycolipid is able to protect normal tissue from this type of chemical induced damage.

NT Factor^TM   phosphoglycolipid contains high concentrations of lysolecithins.

 When NT Factor^TM phosphoglycolipid was analyzed in our laboratory and its composition compared to that of the parent soy-derived material from which it was extracted, we found that NT Factor^TM phosphoglycolipid contains substantially more phosphatidylcholine than the parent material.  Although the fatty acid composition of the phosphatidylcholine from NT Factor^TM   phosphoglycolipid was not different from that of the parent compound, by virtue of concentrating the phosphatidylcholine the extraction process also concentrated polyunsatured phosphatidylcholine.  The greatest difference between the preparations was that NT Factor^TM phosphoglycolipids had over 6 times the lysolecithin content of the parent compound.  This suggests that any unique biological activity of NT Factor^TM may be due in part to its lysolecithin content, either acting alone or in concert with other of its components.

Lysolecithin derivatives disrupt cancer cells at concentrations that do not affect normal cells.

Lysolecithin-like molecules are selectively cytotoxic to cancer cells in vitro (Hoffman 1986, Harmann 1986, Berger 1984).  Such compounds inhibit HL60 leukemic cells at a dosage that has no effect on normal human marrow cells, the tissue from which the leukemic cells are derived.  Normal cells were able to tolerate 4 times higher dosage than the leukemic cells during 24 hours incubation with the phospholipid preparation (Berdel 1986).  There was up to a 5-fold difference in sensitivity between the normal and tumor cells with breast, ovarian, and lung cancer cells, as well as with mesothelioma cells (Namba 1993).

 In summary, some cancerous cells are able to deplete normal cells of phospholipids, causing a degradation in function, and possibly leading to lethargy.  NT Factor^TM  phosphoglycolipid is a very rich source of phospholipids, and also contains high levels of lysolecithin.  Lysolecithin-like molecules are able to inhibit tumors at doses that do not affect normal cells.

REFERENCES:

Aso Y, Kujita K, Tajima A, Suzuki K, Yokoyama M. (1981) Acta Urol. Jpn. 27:1345-1349.

Berdel WE, Von Hoff DD, Unger C, Schick HD, Fink U, Reichert A, Eible H, Rastetter J. (1986) Ether lipid derivatives: Antineoplastic activity in vitro and the structure-activity relationship.  Lipids 21:301-304.

Bergelson LD, Dyatlovitskaya EV, Sorokina IV, Gorkova IB.  (1974) Biochim. Biophys. Acta 360-361.

Bergelson LD, (1972) Tumor lipids. Prog. Chem. Fats Lipids 13:1

Berridge MJ. Irvine RF. (1989) Inositol phosphates and cell signaling. Nature 341:197

Blusztain JK, Liscovitch M, Mauron C, Richardson UI, Wurtman RJ. (1987) Phosphatidylcholine as a precursor of choline for acetylcholine synthesis J. Neural Trans. 24:247

Bogguest WA. (1973) Adv. Tumoyur Prev. Detect. Charact (Charact. Hum. Tumours, Proc. Int. Symp., 5^th)1974: 279-289

Burlakova EB, Palmina NP, Maltseva EL. (1991) In Vigo-Pelfrey C (ed.), Membrane Lipid Oxidation III. Boca Raton, Florida: CRC Press, pp. 209-237

Burlakova KJ, Molochkina EM, Palmina NP. (1980) In Weber G (ed.) Advances in Enzyme Regulation. New York: Pergamon Press, vol 15, pp.163-179.

Cohen EL, Wurtman RJ. (1976) Science 19:561

Conlay LA, Wurtman RJ, Blusztajn K, Coviella IL, Maher TJ, Evoniuk GE. (1986) New Engll. J. Med.  175:892

Harmann DB, Heumann HA. (1986) Cytotoxic ether phospholipids. Different affinities to Lysophosphocholine acytransferases in sensitive and resistant cells.  J. Biol. Chem. 261:7742-7747.

Haubrich DR, Wang PF, Chippendale T, Proctor E. (1976) J. Neurochem. 27:1305.

Hietanen E, Punnonen K, Punnonen, R, Auvinen O. (1986) Carcinogenesis 7:1965-1969.

Hoffman DR, Hoffman LH, Synder F. (1986) Cytotoxicity and metabolism of alkyl phopholipid analogues in neoplastic cells. Cancer Res. 46:5803-5809.

Hostetler KJ, Zenner BD, Morris HP. (1976) Biochim. Biophys, Acta 441:231

Leung BS, Sun GY. (1976) Proc. Soc. Exp. Biol. Med. 152: 671-676.

Margolis B, Zilberstein A, Franks C, Felder S, Kremer S, Ullrich A, Rhee SG, Skorecki K, Schlessinger J.(1990) Effect of phospholipase-C-coverexpression on PDGF induced second messengers and mitogenesis.

Science 248:607

Michell RH. (1998) Phosphoinositides and inositol phosphates. Biochem. Soc. Trans. 17: 1.

Namba Y. (1993) Medical applications of phospholipid. In Cevc G (ed.), Phospholipids Handbook.  New York: Marcel Dekker, Inc., p 892.

Nikolasev V, Lazar G, Karady I. (1972) Kiserl. Orvostud. 24:465-469

Nordoy A, Goodnight SH. (1990) Dietary lipids and thrombosis.  Arteriosclerosis 10:149

Palmina NP. (1995) Phosphatidylcholine exchange in membranes of normal and tumor tissues.  In Cevc G,

Paltauf F (eds.). Phospholipids:  Characterization, Metabolism, and Novel Biological Applications.

Chamaign, Illinois: AOCS Press, pp. 311-318.

Poorthuis Ben JHM, van der Krift TP, Teerlink T, Akeroyd R, Hjostetler KW, Wirtz KWA. (1980) Biochim.

Biophys. Acta 600:376.

Rana Rs, Hokin LE. (1990) Role of phosphoinositides in transmembrane signaling, Phys. Rev. 70:  115.

Snyder F, Lee TC, Blank ML. (1989) Platelet activating factor and related ether lipid mediators.  Ann. NY Acad. Sci. 568:35.

Spangler M, Coetzee ML, Katyal SL, Morris HP, Ove  P. (1975) Cancer Res. 35:3131.

Spector A A, Mathur SN, Kaduce TL, Hyman BT. (1981) Lipid nutrition and metabolism of cultured  mammalian cells.  Prog. Lipid Res. 19:155.

Spector AA, Yorek MA.  (1985) Membrane lipid compositin and cellular function.  J. Lipid Res. 26:10105.

Sun GY, Foudin LL. (1985) Phospholipid composition and metabolism in the developing and aging nervous System.  In Eichberg J (ed.), Phospolipids in Nervous Tissue.  New York: Wiley, p. 79.

Takenaka R, Inoue M, Hori T, Okuyama H. (1983) Biochim.  Biophys. Acta 754: 28-37.

Tan W C, Chapman C, Takatori T, Privett OS, (1975) Lipids 10:70-74.

Von Allwörden HN, Horn S, Feldheim W. (1995) The influence of lecithin on the performance and the recovery process of endurance athletes.  In Cevc G,  Paltauf F (eds.) Phospholipids: Characterization, Metabolism, and Novel Biological Applications.  Champaign, Illlinois:  AOCS Press, pp. 319-325.

Von Allwörden HN, Horn S, Kahl J, Feldheim W. (1993) Eur. J. Appl. Physiol. 67: 87.

Yorek MA (1993) Biological distribution.  In Cevc G (ed.), Phospholipids Handbook. New York: Marcel Dekker, Inc., p. 760.

Cancerous cells of tumors import phospholipids from normal cells

Reduced levels of phospho-lipids in normal cells can limit metabolic activity and limit available energy

The rate of phospholipid accumulation in cancer cells is independent of exogenous supply

NT Factor^TM phospho-glycolipid improves cell maintenance and metabolic activity of normal cells

NT Factor^TM   phospho-glycolipid contains high concentrations of lysolecithins

Lysolecithin derivatives disrupt cancer cells at concentrations that do not affect normal cells

REFERENCES