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Increased phospholipase D activity in human breast cancer

  • Original Paper
  • Clinical Oncology
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

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.

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Abbreviations

PtdCho :

phosphatidylcholine

PLD :

phospholipase D

GroPCho:

sn-glycero-3-phosphocholine

TLC :

thin-layer chromatography

PtdOEt :

phosphatidylethanol

References

  • Bocckino SB, Blackmore PF, Wilson PB, Exton JH (1987) Phosphatidate accumulation in hormone-treated hepatocytes via a phospholipase D mechanism. J Biol Chem 262:15309–15315

    PubMed  Google Scholar 

  • Bocckino SB, Wilson PB, Exton JH (1991) Phosphatidate-dependent protein phosphorylation. Proc Natl Acad Sci USA 88:6210–6213

    PubMed  Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgam quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    PubMed  Google Scholar 

  • Brown HA, Gutowski S, Moomaw CR, Slaughter C, Sternweis PC (1993) ADP-ribosylation factor, a small GTP-dependent regulatory protein, stimulates phospholipase D activity. Cell 75:1137–1144

    PubMed  Google Scholar 

  • Chalifour R, Kanfer JN (1982) Fatty acid activation and temperature perturbation of rat brain microsomal phospholipase D. J Neurochem 39:299–305

    PubMed  Google Scholar 

  • Chuang TH, Bohl BP, Bokoch GM (1993) Biologically active lipids are regulators of Rac-GDI complexation. J Biol Chem 268:26206–26211

    PubMed  Google Scholar 

  • Cockcroft S, Thomas GMH, Fensome A, Geny B, Cunningham E, Gout I, Hiles I, Totty NF, Truong O, Hsuan JJ (1994) Phospholipase D: a downstream effector of ARF in granulocytes. Science 263:523–526

    PubMed  Google Scholar 

  • Exton JH (1994) Phosphatidylcholine breakdown and signal transduction. Biochim Biophys Acta 1212:26–42

    PubMed  Google Scholar 

  • Gratas C, Powis G (1993) Inhibition of phospholipase D by agents that inhibit cell growth. Anticancer Res 13:1239–1244

    PubMed  Google Scholar 

  • Guillem JG, O'Brian CA, Fitzer CJ, Forde KA, LoGerfo P, Treat M, Weinstein IB (1987) Altered levels of protein kinase C and Ca2+-dependent protein kinases in human colon carcinomas. Cancer Res 47:2036–2039

    PubMed  Google Scholar 

  • Hagiwara M, Hachiya T, Watanabe M, Usuda N, Iida F, Tamai K, Hidaka H (1990) Assessment of protein kinase C isozymes by enzyme immunoassay and overexpression of type II in thyroid adenocarcinoma. Cancer Res 50:5515–5519

    PubMed  Google Scholar 

  • Imamura F, Horai T, Mukai M, Shinkai K, Sawada M, Akedo H (1993) Induction of in vitro tumor cell invasion of cellular monolayers by lysophosphatidic acid or phospholipase D. Biochem Biophys Res Commun 193:497–503

    PubMed  Google Scholar 

  • Jackowski S, Rock CO (1989) Stimulation of phosphatidylinositol 4,5-bisphosphate phospholipase C activity by phosphatidic acid. Arch Biochem Biophys 268:516–524

    PubMed  Google Scholar 

  • Jenkins GH, Fisette PL, Anderson RA (1994) Type I Phosphatidylinositol 4-phosphate 5-kinase isoforms are specifically stimulated by phosphatidic acid. J Biol Chem 269:11547–11554

    PubMed  Google Scholar 

  • Khan WA, Blobe GC, Richards AL, Hannun YA (1994) Identification, partial purification, and characterization of a novel phospholipid-dependent and fatty acid-activated protein kinase from human platelets. J Biol Chem 269:9729–9735

    PubMed  Google Scholar 

  • Kishimoto A, Takai Y, Mori T, Kikkawa U, Nishizuka Y (1980) Activation of calcium and phospholipid-dependent protein kinase by diacylglycerol, its possible relation to phosphatidylinositol turnover. J Biol Chem 255:2273–2276

    PubMed  Google Scholar 

  • Kobayashi M, Kanfer JN (1987) Phosphatidylethanol formation via transphosphatidylation by rat brain synaptosomal phospholipase D. J Neurochem 48:1597–1603

    PubMed  Google Scholar 

  • Kopp R, Noelke B, Sauter G, Schildberg FW, Paumgartner G, Pfeiffer A (1991) Altered protein kinase C activity in biopsies of human colonic adenomas and carcinomas. Cancer Res 51:205–210

    PubMed  Google Scholar 

  • Malcolm KC, Ross AH, Qiu RG, Symons M, Exton JH (1994) Activation of rat liver phospholipase D by the small GTP-binding protein RhoA. J Biol Chem 269:25951–25954

    PubMed  Google Scholar 

  • Matozaki T, Williams JA (1989) Multiple sources of 1,2-diacylglycerol in isolated rat pancreatic acini stimulated by cholecystokinin. Involvement of phosphatidylinositol bisphosphate and phosphatidylcholine hydrolysis. J Biol Chem 264:14729–14734

    PubMed  Google Scholar 

  • Moolenaar WH, Kruijer W, Tilly BC, Verlaan I, Bierman AJ, Laat SW de (1986) Growth factor-like action of phosphatidic acid. Nature 323:171–173

    PubMed  Google Scholar 

  • Moritz A, De Graan PNE, Gispen WH, Wirtz KWA (1992) Phosphatidic acid is a specific activator of phosphatidylinositol-4-phosphate kinase. J Biol Chem 267:7207–7210

    PubMed  Google Scholar 

  • Mufson RA, Okin E, Weinstein IB (1981) Phorbol esters stimulate the rapid release of choline from prelabelled Cells. Carcinogenesis 2:1095–1102

    PubMed  Google Scholar 

  • Murayama T, Ui M (1987) Phosphatidic acid may stimulate membrane receptors mediating adenylate cyclase inhibition and phospholipid breakdown in 3T3 fibroblasts. J Biol Chem 262:5522–5529

    PubMed  Google Scholar 

  • Nakanishi H, Exton JH (1992) Purification and characterization of the ζ isoform of protein kinase C from bovine kidney. J Biol Chem 267:16347–16354

    PubMed  Google Scholar 

  • Nishizuka Y (1992) Intracellular signaling by hydrolysis of phospholipids and activation of protein kinase C. Science 258:607–614

    PubMed  Google Scholar 

  • O'Brian CA, Vogel VG, Singletary SE, Ward NE (1989) Elevated protein kinase C expression in human breast tumor biopsies relative to normal breast tissue. Cancer Res 49:3215–3217

    PubMed  Google Scholar 

  • Okamura S, Yamashita S (1994) Purification and characterization of phosphatidylcholine phospholipase D from pig lung. J Biol Chem 269:31207–31213

    PubMed  Google Scholar 

  • Pai JK, Siegel MI, Egan RW, Billah MM (1988) Phospholipase D catalyzes phospholipid metabolism in chemotactic peptidestimulated HL-60 granulocytes. J Biol Chem 263:12472–12477

    PubMed  Google Scholar 

  • Sato T, Hashizume T, Fujii T (1992) Preferential activation of phospholipase A2 by low concentrations of phosphatidic acid with long-chain fatty acids in rabbit platelets. J Biochem (Tokyo) 112:756–761

    Google Scholar 

  • Schnitt SJ, Connolly JL, Harris JR, Hellman S, Cohen, RB (1984) Pathologic predictors of early local recurrence in stage I and II breast cancer treated by primary radiation therapy. Cancer 53:1049–1057

    PubMed  Google Scholar 

  • Siegmann DW (1987) Stimulation of quiescent 3T3 cells by phosphatidic acid-containing liposomes. Biochem Biophys Res Commun 145:228–233

    PubMed  Google Scholar 

  • Takai Y, Kishimoto A, Kikkawa U, Mori T, Nishizuka Y (1979) Unsaturated diacylglycerol as a possible messenger for the activation of calcium-activated, phospholipid-dependent protein kinase system. Biochem Biophys Res Commun 91:1218–1224

    PubMed  Google Scholar 

  • Tettenborn CS, Mueller GC (1988) 12-O-Tetradecanoylphorbol-13-acetate activates phosphatidylethanol and phosphatidylglycerol synthesis by phospholipase D in cell lysates. Biochem Biophys Res Commun 155:249–255

    PubMed  Google Scholar 

  • Tsai MH, Yu CL, Wei FS, Stacey DW (1989) The effect of GTPase activating protein upon Ras is inhibited by mitogenically responsive lipids. Science 243:522–526

    PubMed  Google Scholar 

  • Van Blitterswijk WJ, Hilkmann H, Widt J de, Bend RL van der (1991) Phospholipid metabolism in bradykinin-stimulated human fibroblasts. I. Biphasic formation of diacylglycerol from phosphatidylinositol and phosphatidylcholine, controlled by protein kinase C. J Biol Chem 166:10337–10343

    Google Scholar 

  • Van Corven EJ, Groenink A, Jalink K, Eichholtz T, Moolenaar WH (1989) Lysophosphatidate-induced cell proliferation: identification and dissection of signaling pathways mediated by G proteins. Cell 59:45–54

    PubMed  Google Scholar 

  • Wright TM, Rangan LA, Shin HS, Raben DM (1988) Kinetic analysis of 1,2-diacylglycerol mass levels in cultured fibroblasts. Comparison of stimulation by α-thrombin and epidermal growth factor. J Biol Chem 263:9374–9380

    PubMed  Google Scholar 

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Authors and Affiliations

  1. First Department of Surgery, Gunma University School of Medicine, 371, Maebashi, Japan

    Nobuyuki Uchida & Yukio Nagamachi

  2. Department of Biochemistry, Gunma University School of Medicine, 371, Maebashi, Japan

    Shin-ichi Okamura & Satoshi Yamashita

Authors
  1. Nobuyuki Uchida
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  2. Shin-ichi Okamura
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  3. Yukio Nagamachi
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  4. Satoshi Yamashita
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Corresponding author

Correspondence to Satoshi Yamashita.

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This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture, Japan

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Uchida, N., Okamura, Si., Nagamachi, Y. et al. Increased phospholipase D activity in human breast cancer. J Cancer Res Clin Oncol 123, 280–285 (1997). https://doi.org/10.1007/BF01208639

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  • DOI: https://doi.org/10.1007/BF01208639

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