Advertisement

Breast Cancer Research and Treatment

, Volume 54, Issue 2, pp 135–146 | Cite as

De‐regulation of GRP stress protein expression in human breast cancer cell lines

  • Gadi Gazit
  • Jun Lu
  • Amy S. Lee
Article

Abstract

The stress‐inducible glucose regulated proteins (GRPs), a class of calcium‐binding molecular chaperones localized in the endoplasmic reticulum, have been implicated in the development of tumorigenicity, drug resistance, and cytotoxic immunology. This study investigates the expression pattern of GRP94 and GRP78 in a panel of breast carcinoma cell lines, as compared to two independently derived normal human breast epithelial cell lines. Here we report that a 3‐ to 5‐fold increase in the basal level of the GRP94 protein was observed in all five breast carcinoma cell lines examined. The increase was independent of either the p53 or estrogen receptor status of the breast carcinomas. In carcinoma cells deprived of glucose, mimicking the conditions in poorly vascularized solid tumors, up to 9‐fold induction of GRP94 was observed relative to the basal level expressed in a normal breast epithelial cell line. Interestingly, while the majority of the breast cancer cell lines can respond to tunicamycin‐ and thapsigargin‐induced stress by increasing the steady state levels of grp94 and grp78 transcripts, the induction at the GRP protein level is variable and does not always correspond with the transcript level. Further, we discovered that one of the human breast carcinoma cell lines, MCF‐7, has specifically lost its ability to respond to tunicamycin stress.

stress protein expression human breast cancer 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Shiu RP, Pastan IH: Properties and purification of a glucoseregulated protein from chick embryo fibroblasts. Biochim Biophys Acta 576: 141–150, 1979Google Scholar
  2. 2.
    Gething MJ, Sambrook J: Protein folding in the cell. Nature 355: 33–45, 1992Google Scholar
  3. 3.
    Lee AS: Mammalian stress response: Induction of the glucose-regulated protein family. Curr Opin Cell Biol 4: 267–273, 1992Google Scholar
  4. 4.
    Little E, Ramakrishnan M, Roy B, Gazit G, Lee AS: The glucose-regulated proteins (GRP78 and GRP94): Functions, gene regulation, and applications. Crit Rev Eukaryot Gene Expr 4: 1–18, 1994Google Scholar
  5. 5.
    Sciandra JJ, Subjeck JR, Hughes CS: Induction of glucoseregulated proteins during anaerobic exposure and of heatshock proteins after reoxygenation. Proc Natl Acad SciUSA 81: 4843–4847, 1984Google Scholar
  6. 6.
    Lee AS: Coordinated regulation of a set of genes by glucose and calcium ionophores in mammalian cells. Trends Biochem Sci 12: 20–23, 1987Google Scholar
  7. 7.
    Patierno SR, Tuscano JM, Kim KS, Landolph JR, Lee AS: Increased expression of the glucose-regulated gene encoding the Mr 78,000 glucose-regulated protein in chemically and radiation-transformedC3H10T1/2 mouse embryo cells. Cancer Res 47: 6220–6224, 1987Google Scholar
  8. 8.
    Cai JW, Henderson BW, Shen JW, Subjeck JR: Induction of glucose-regulated proteins during growth of a murine tumor. J Cell Physiol 154: 229–237, 1993Google Scholar
  9. 9.
    Gazit G, Kane SE, Nichols P, Lee AS: Use of the stressinducible grp78/BiP promoter in targeting high level gene expression in fibrosarcoma in vivo. Cancer Res 55: 1660–1663, 1995Google Scholar
  10. 10.
    Thastrup O, Cullen PJ, Drobak BK, Hanley MR, Dawson AP: Thapsigargin, a tumor promoter, discharges intracellular Ca2+ stores by specific inhibition of the endoplasmic reticulum Ca2+-ATPase. Proc Natl Acad Sci USA 87: 2466–2470, 1990Google Scholar
  11. 11.
    Li LJ, Li X, Ferrario A, Rucker N, Liu ES, Wong S, Gomer CJ, Lee AS: Establishment of a Chinese hamster ovary cell line that expresses grp78 antisense transcripts and suppresses A23187 induction of both GRP78 and GRP94. J Cell Physiol 153: 575–582, 1992Google Scholar
  12. 12.
    Little E, Lee AS: Generation of a mammalian cell line defi-cient in glucose-regulated protein stress induction through targeted ribozyme driven by a stress-inducible promoter. J Biol Chem 270: 9526–9534, 1995Google Scholar
  13. 13.
    Little E, Tocco G, Baudry M, Lee AS, Schreiber SS: Induction of glucose-regulated protein (glucose-regulated protein 78/BiP and glucose-regulated protein 94) and heat shock protein 70 transcripts in the immature rat brain following status epilepticus. Neuroscience 75: 209–219, 1996Google Scholar
  14. 14.
    Liu H, Bowes RC III, van de Water B, Sillence C, Nagelkerke JF, Stevens JL: Endoplasmic reticulum chaperones GRP78 and calreticulin prevent oxidative stress, Ca2+ disturbances, and cell death in renal epithelial cells. J Biol Chem 272: 21751–21759, 1997Google Scholar
  15. 15.
    Morris JA, Dorner AJ, Edwards CA, Hendershot LM, Kaufman RJ: Immunoglobulin binding protein (BiP) function is required to protect cells from endoplasmic reticulum stress but is not required for the secretion of selective proteins. J Biol Chem 272: 4327–4334, 1997Google Scholar
  16. 16.
    Sugawara S, Nowicki M, Xie S, Song HJ, Dennert G: Effects of stress on lysability of tumor targets by cytotoxic T cells and tumor necrosis factor. J Immunol 145: 1991–1998, 1990Google Scholar
  17. 17.
    Sugawara S, Takeda K, Lee A, Dennert G: Suppression of stress protein GRP78 induction in tumor B/C10ME eliminates resistance to cell mediated cytotoxicity. Cancer Res 53: 6001–6005, 1993Google Scholar
  18. 18.
    Jamora C, Dennert G, Lee AS: Inhibition of tumor progression by suppression of stress protein GRP78/BiP induction in fibrosarcoma B/C10ME. Proc Natl Acad Sci USA 93:7690–7694, 1996Google Scholar
  19. 19.
    Menoret A, Meflah K, Le Pendu J: Expression of the100 kDa glucose-regulated protein (GRP100/endoplasmin) is associated with tumorigenicity in a model of rat colon adenocarcinoma. Int J Cancer 56: 400–405, 1994Google Scholar
  20. 20.
    Shen J, Hughes C, Chao C, Cai J, Bartels C, Gessner T, Subjeck J: Coinduction of glucose-regulated proteins and doxorubicin resistance in Chinese hamster cells. Proc Natl Acad Sci USA 84: 3278–3282, 1987Google Scholar
  21. 21.
    Hughes CS, Shen JW, Subjeck JR: Resistance to etoposide induced by three glucose-regulated stresses in Chinese hamster ovary cells. Cancer Res 49: 4452–4454, 1989Google Scholar
  22. 22.
    Vichi PJ, Tritton TR: Protection from adriamycin cytotoxicity in L1210 cells by brefeldin A. Cancer Res 53: 5237–5243, 1993Google Scholar
  23. 23.
    Chatterjee S, Cheng MF, Berger RB, Berger SJ, Berger NA: Effect of inhibitors of poly(ADP-ribose) polymerase on the induction of GRP78 and subsequent development of resistance to etoposide. Cancer Res 55: 868–873, 1995Google Scholar
  24. 24.
    Gomer CJ, Ferrario A, Rucker N, Wong S, Lee AS: Glucose regulated protein induction and cellular resistance to oxidative stress mediated by porphyrin photosensitization. Cancer Res 51: 6574–6579, 1991Google Scholar
  25. 25.
    Ciocca DR, Fuqua SA, Lock-Lim S, Toft DO, Welch WJ, McGuire WL: Response of human breast cancer cells to heat shock and chemotherapeutic drugs. Cancer Res 52: 3648–3654, 1992Google Scholar
  26. 26.
    Ciocca DR, Clark GM, Tandon AK, Fuqua SA, Welch WJ, McGuire WL: Heat shock protein HSP70 in patients with axillary lymph node-negative breast cancer: Prognostic implications. J Natl Cancer Inst 85: 570–574, 1993Google Scholar
  27. 27.
    Oesterreich S, Weng CN, Qiu M, Hilsenbeck SG, Osborne CK, Fuqua SA: The small heat shock protein HSP27 is correlated with growth and drug resistance in human breast cancer cell lines. Cancer Res 53: 4443–4448, 1993Google Scholar
  28. 28.
    Chavany C, Mimnaugh E, Miller P, Bitton R, Nguyen P, Trepel J, Whitesell L, Schnur R, Moyer J, Neckers L: p185erbB2 binds to GRP94 in vivo. Dissociation of the p185erbB2/GRP94 heterocomplex by benzoquinone ansamycins precedes depletion of p185erbB2. J Biol Chem 271: 4974–4977, 1996Google Scholar
  29. 29.
    Lee AS, Delegeane AM, Baker V, Chow PC:Transcriptional regulation of two genes specifically induced by glucose starvation in a hamster mutant fibroblast cell line. J Biol Chem 258: 597–603, 1983Google Scholar
  30. 30.
    Tso JY, Sun XH, Kao TH, Reece KS, Wu R: Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: Genomic complexity and molecular evolution of the gene. Nucleic Acids Res 13: 2485–2502, 1985Google Scholar
  31. 31.
    deFazio A, Chiew YE, Donoghue C, Lee CS, Sutherland RL: Effect of sodium butyrate on estrogen receptor and epidermal growth factor receptor gene expression in human breast cancer cell lines. J Biol Chem 267: 18008–18012, 1992Google Scholar
  32. 32.
    Ogretmen B, Ratajczak H, Gendel SM, Stark BC: Effects of sodium saccharin and linoleic acid on mRNA levels of Her2/neu and p53 in a human breast epithelial cell line. Cancer Lett 102: 91–99, 1996Google Scholar
  33. 33.
    Shao ZM, Dawson MI, Li XS, Rishi AK, Sheikh MS, Han QX, Ordonez JV, Shroot B, Fontana JA: p53 independent G0/G1 arrest and apoptosis induced by a novel retinoid in human breast cancer cells. Oncogene 11: 493–504, 1995Google Scholar
  34. 34.
    Nakshatri H, Bhat-Nakshatri P, Martin DA, Goulet RJ Jr, Sledge GW Jr: Constitutive activation of NF-κB during progression of breast cancer to hormone-independent growth. Mol Cell Biol 17: 3629–3639, 1997Google Scholar
  35. 35.
    Kim YK, Kim KS, Lee AS: Regulation of the glucoseregulated protein genes by beta-mercaptoethanol requires de novo protein synthesis and correlates with inhibition of protein glycosylation. J Cell Physiol 133: 553–559, 1987Google Scholar
  36. 36.
    Watowich SS, Morimoto RI: Complex regulation of heat shock-and glucose-responsive genes in human cells. Mol Cell Biol 8: 393–405, 1988Google Scholar
  37. 37.
    Li WW, Alexandre S, Cao X, Lee AS: Transactivation of the grp78 promoter by Ca2+ depletion. A comparative analysis with A23187 and the endoplasmic reticulum Ca2C-ATPase inhibitor thapsigargin. J Biol Chem 268: 12003–12009, 1993Google Scholar
  38. 38.
    Falany JL, Falany CN: Expression of cytosolic sulfotransferases in normal mammary epithelial cells and breast cancer cell lines. Cancer Res 56: 1551–1555, 1996Google Scholar
  39. 39.
    Williard R, Benz CC, Baxter JD, Kushner P, Hunt CA: Paradoxical production of target protein using antisense RNA expression vectors. Gene 149: 21–24, 1994Google Scholar
  40. 40.
    Amellem O, Stokke T, Sandvik JA, Smedshammer L, Pettersen EO: Hypoxia-induced apoptosis in human cells with normal p53 status and function, without any alteration in the nuclear protein level. Exp Cell Res 232: 361–370, 1997Google Scholar
  41. 41.
    Elstner E, Linker-Israeli M, Said J, Umiel T, de Vos S, Shintaku IP, Heber D, Binderup L, Uskokovic M, Koeffler HP: 20-epi-vitamin D3 analogues: A novel class of potent inhibitors of proliferation and inducers of differentiation of human breast cancer cell lines. Cancer Res 55: 2822–2830, 1995Google Scholar
  42. 42.
    Hoffman R: Potent inhibition of breast cancer cell lines by the isoflavonoid kievitone: Comparison with genistein. Biochem Biophys Res Commun 211: 600–606, 1995Google Scholar
  43. 43.
    Lee AS: The accumulation of three specific proteins related to glucose-regulated proteins in a temperature-sensitive hamster mutant cell line K12. J Cell Physiol 106: 119–125, 1981Google Scholar
  44. 44.
    McCormick TS, McColl KS, Distelhorst CW: Mouse lymphoma cells destined to undergo apoptosis in response to thapsigargin treatment fail to generate a calcium-mediated grp78/grp94 stress response. J Biol Chem 272: 6087–6092, 1997Google Scholar
  45. 45.
    Kozutsumi Y, Segal M, Normington K, Gething MJ, Sambrook J: The presence of malfolded proteins in the endoplasmic reticulum signals the induction of glucose-regulated proteins. Nature 332: 462–464, 1988Google Scholar
  46. 46.
    Melnick J, Dul JL, Argon Y: Sequential interaction of the chaperones BiP and GRP94 with immunoglobulin chains in the endoplasmic reticulum. Nature 370: 373–375, 1994Google Scholar
  47. 47.
    Bruneau N, Lombardo D: Chaperone function of a GRP 94-related protein for folding and transport of the pancreatic bile salt-dependent lipase. J Biol Chem 270: 13524–13533,1995Google Scholar
  48. 48.
    Ramakrishnan M, Tugizov S, Pereira L, Lee AS: Conformation-defective herpes simplex virus 1 glycoprotein B activates the promoter of the grp94 gene that codes for the 94 kDa stress protein in the endoplasmic reticulum. DNA Cell Biol 14: 373–384, 1995Google Scholar
  49. 49.
    Li Z, Srivastava PK: Tumor rejection antigen gp96/grp94 is an ATPase: Implications for protein folding and antigen presentation. EMBO J 12: 3143–3151, 1993Google Scholar
  50. 50.
    Sambrook JF: The involvement of calcium in transport of secretory proteins from the endoplasmic reticulum. Cell 61:197–199, 1990Google Scholar
  51. 51.
    Rutherford SL, Zuker CS: Protein folding and the regulation of signaling pathways. Cell 79: 1129–1132, 1994Google Scholar
  52. 52.
    Dorner AJ, Wasley LC, Kaufman RJ: Overexpression of GRP78 mitigates stress induction of glucose regulated proteins and blocks secretion of selective proteins in Chinese hamster ovary cells. EMBO J 11: 1563–1571, 1992Google Scholar
  53. 53.
    Kiang JG, Gist ID, Tsokos GC: 17 beta-estradiol-induced increase in glucose-regulated proteins 78 kDa and 94 kDa protects human breast cancer T47-D cells from thermal injury. Chin J Physiol 40: 213–219, 1997Google Scholar
  54. 54.
    Poola I, Kiang JG: The estrogen-inducible transferrin receptor-like membrane glycoprotein is related to stressregulated proteins. J Biol Chem 269: 21762–21769, 1994Google Scholar
  55. 55.
    Hentze MW: Translational regulation: Versatile mechanisms for metabolic and developmental control. Curr Opin Cell Biol 3: 393–398, 1995Google Scholar
  56. 56.
    Lazaris-Karatzas A, Montine KS, Sonenberg N: Malignant transformation by a eukaryotic initiation factor subunit that binds to mRNA 50 cap. Nature 345: 544–547, 1990Google Scholar
  57. 57.
    Shantz LM, Pegg AE: Overproduction of ornithine decarboxylase caused by relief of translational repression is associated with neoplastic transformation. Cancer Res 54: 2313–2316, 1994Google Scholar
  58. 58.
    Koromilas AE, Lazaris-Karatzas A, Sonenberg N: mRNAs containing extensive secondary structure in their 5' noncoding region translate efficiently in cells overexpressing initiation factor eIF-4E. EMBO J 11: 4153–4158, 1992Google Scholar
  59. 59.
    Ting J: Thesis. University of Southern California, Los Angeles, 1987Google Scholar
  60. 60.
    Price BD, Calderwood SK: Gadd45 and Gadd153 messenger RNA levels are increased during hypoxia and after exposure of cells to agents which elevate the levels of the glucose-regulated proteins. Cancer Res 52: 3814–3817, 1992Google Scholar
  61. 61.
    Cao X, Zhou Y, Lee AS: Requirement of tyrosine and serine/threonine kinases in the transcriptional activation of the mammalian grp78/BiP promoter by thapsigargin. J Biol Chem 270: 494–502, 1995Google Scholar
  62. 62.
    Zhou Y, Lee AS: Mechanism for the suppression of the mammalian stress response by genistein, an anticancer phytoestrogen from soy. J Natl Cancer Inst 90: 381–388, 1998Google Scholar

Copyright information

© Kluwer Academic Publishers 1999

Authors and Affiliations

  • Gadi Gazit
    • 1
  • Jun Lu
    • 1
  • Amy S. Lee
    • 1
  1. 1.Department of Biochemistry and Molecular Biology, USC/Norris Comprehensive Cancer CenterUniversity of Southern California School of MedicineLos AngelesUSA

Personalised recommendations