Increased phospholipase D activity in human breast cancer

Phospholipase D is believed to play an important role in cell proliferation and tumorigenesis. One of its major functions is to cause a sustained activation of protein kinase C through the primary production of phosphatidic acid from phosphatidylcholine by the enzyme, followed by dephosphorylation forming diacylglycerol. Protein kinase C is known to be activated or translocated in some tumors including breast tumors. In order to examine phospholipase D activity in breast tumors, surgical specimens of human breast tumors were obtained by mastectomy or wide excision, and their phospholipase D activities were assayed by determining the formation of phosphatidylethanol from phosphatidylcholine and ethanol. Phospholipase D activity was predominantly localized in the microsomal fraction of the tumor tissue and markedly stimulated by oleic acid. We observed a significant increase in phospholipase D activity in 17 out of 19 spontaneous human breast tumors as compared to adjacent histologically normal breast tissue. The mean specific activity in the tumors was 52.9±41.8 (SD) pmol min−1 mg protein−1 whereas the value for the normal breast tissue was 34.0±36.2 (SD) pmol min−1 mg protein−1 (P<0.01; paired Wilcoxon's rank-sum test). The mean tumor/normal activity ratio was 2.37. Among prognostic factors, the nuclear grade, evaluated according to Schnitt et al., was found to be correlated with the activity ratio. Our results suggest a role for phospholipase D in human breast tumors. An elevation in phospholipase D activity is useful as a potential marker for malignant disease in the breast.

[1]  J. Kanfer,et al.  Phosphatidylethanol Formation via Transphosphatidylation by Rat Brain Synaptosomal Phospholipase D , 1987, Journal of neurochemistry.

[2]  I. Weinstein,et al.  Phorbol esters stimulate the rapid release of choline from prelabelled cells. , 1981, Carcinogenesis.

[3]  Y. Hannun,et al.  Identification, partial purification, and characterization of a novel phospholipid-dependent and fatty acid-activated protein kinase from human platelets. , 1994, The Journal of biological chemistry.

[4]  S. Okamura,et al.  Purification and characterization of phosphatidylcholine phospholipase D from pig lung. , 1994, The Journal of biological chemistry.

[5]  K. Wirtz,et al.  Phosphatidic acid is a specific activator of phosphatidylinositol-4-phosphate kinase. , 1992, The Journal of biological chemistry.

[6]  C. Yu,et al.  The effect of GTPase activating protein upon ras is inhibited by mitogenically responsive lipids. , 1989, Science.

[7]  S. Schnitt,et al.  Pathologic predictors of early local recurrence in stage I and II breast cancer treated by primary radiation therapy , 1984, Cancer.

[8]  I. Gout,et al.  Phospholipase D: a downstream effector of ARF in granulocytes. , 1994, Science.

[9]  W. V. van Blitterswijk,et al.  Phospholipid metabolism in bradykinin-stimulated human fibroblasts. I. Biphasic formation of diacylglycerol from phosphatidylinositol and phosphatidylcholine, controlled by protein kinase C. , 1991, The Journal of biological chemistry.

[10]  J. Exton Phosphatidylcholine breakdown and signal transduction. , 1994, Biochimica et biophysica acta.

[11]  G M Bokoch,et al.  Biologically active lipids are regulators of Rac.GDI complexation. , 1993, The Journal of biological chemistry.

[12]  J. Kanfer,et al.  Fatty Acid Activation and Temperature Perturbation of Rat Brain Microsomal Phospholipase D , 1982, Journal of neurochemistry.

[13]  S. Singletary,et al.  Elevated protein kinase C expression in human breast tumor biopsies relative to normal breast tissue. , 1989, Cancer research.

[14]  Kees Jalink,et al.  Lysophosphatidate-induced cell proliferation: Identification and dissection of signaling pathways mediated by G proteins , 1989, Cell.

[15]  T. Sato,et al.  Preferential activation of phospholipase A2 by low concentrations of phosphatidic acid with long-chain fatty acids in rabbit platelets. , 1992, Journal of biochemistry.

[16]  P. Blackmore,et al.  Phosphatidate accumulation in hormone-treated hepatocytes via a phospholipase D mechanism. , 1987, The Journal of biological chemistry.

[17]  D. Siegmann,et al.  Stimulation of quiescent 3T3 cells by phosphatidic acid-containing liposomes. , 1987, Biochemical and biophysical research communications.

[18]  F. Schildberg,et al.  Altered protein kinase C activity in biopsies of human colonic adenomas and carcinomas. , 1991, Cancer research.

[19]  小林 睦博 Phosphatidylethanol formation via transphosphatidylation by rat brain synaptosomal phospholipase D , 1990 .

[20]  M. M. Bradford A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[21]  R. Egan,et al.  Phospholipase D catalyzes phospholipid metabolism in chemotactic peptide-stimulated HL-60 granulocytes. , 1988, The Journal of biological chemistry.

[22]  J. Exton,et al.  Purification and characterization of the zeta isoform of protein kinase C from bovine kidney. , 1992, The Journal of biological chemistry.

[23]  Y. Nishizuka,et al.  Unsaturated diacylglycerol as a possible messenger for the activation of calcium-activated, phospholipid-dependent protein kinase system. , 1979, Biochemical and biophysical research communications.

[24]  M. Symons,et al.  Activation of rat liver phospholipase D by the small GTP-binding protein RhoA. , 1994, The Journal of biological chemistry.

[25]  T. Wright,et al.  Kinetic analysis of 1,2-diacylglycerol mass levels in cultured fibroblasts. Comparison of stimulation by alpha-thrombin and epidermal growth factor. , 1988, The Journal of biological chemistry.

[26]  C. Rock,et al.  Stimulation of phosphatidylinositol 4,5-bisphosphate phospholipase C activity by phosphatidic acid. , 1989, Archives of biochemistry and biophysics.

[27]  M. Ui,et al.  Phosphatidic acid may stimulate membrane receptors mediating adenylate cyclase inhibition and phospholipid breakdown in 3T3 fibroblasts. , 1987, The Journal of biological chemistry.

[28]  I. Verlaan,et al.  Growth factor-like action of phosphatidic acid , 1986, Nature.

[29]  C. S. Tettenborn,et al.  12-O-tetradecanoylphorbol-13-acetate activates phosphatidylethanol and phosphatidylglycerol synthesis by phospholipase D in cell lysates. , 1988, Biochemical and biophysical research communications.

[30]  M. Treat,et al.  Altered levels of protein kinase C and Ca2+-dependent protein kinases in human colon carcinomas. , 1987, Cancer research.

[31]  F. Imamura,et al.  Induction of in vitro tumor cell invasion of cellular monolayers by lysophosphatidic acid or phospholipase D. , 1993, Biochemical and biophysical research communications.

[32]  R. Anderson,et al.  Type I phosphatidylinositol 4-phosphate 5-kinase isoforms are specifically stimulated by phosphatidic acid. , 1994, The Journal of biological chemistry.

[33]  Y. Nishizuka Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. , 1992, Science.

[34]  H. Brown,et al.  ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity , 1993, Cell.

[35]  J. Williams,et al.  Multiple sources of 1,2-diacylglycerol in isolated rat pancreatic acini stimulated by cholecystokinin. Involvement of phosphatidylinositol bisphosphate and phosphatidylcholine hydrolysis. , 1989, The Journal of biological chemistry.

[36]  Y Nishizuka,et al.  Activation of calcium and phospholipid-dependent protein kinase by diacylglycerol, its possible relation to phosphatidylinositol turnover. , 1980, The Journal of biological chemistry.

[37]  M. Hagiwara,et al.  Assessment of protein kinase C isozymes by enzyme immunoassay and overexpression of type II in thyroid adenocarcinoma. , 1990, Cancer research.

[38]  J. Exton,et al.  Phosphatidate-dependent protein phosphorylation. , 1991, Proceedings of the National Academy of Sciences of the United States of America.