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Mechanism of Quercetin-induced suppression and delay of heat shock gene expression and thermotolerance development in HT-29 cells

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Previous studies have shown that a combination of low pH and quercetin (QCT) treatment following heat shock markedly suppresses and delays the expression of heat shock protein genes, particularly the HSP70 gene (Lee et al., Biochem. Biophys. Res. Commun., 186:1121–1128, 1992). The possible mechanism for alteration of gene expression by treatment with QCT at low pH was investigated in human colon carcinoma cells. Cells were heated at 45°C for 15 min and then incubated at 37°C for various times (0–12 h) with QCT (0.05–0.2 mM) at pH 7.4 or 6.5. Gel mobility-shift analysis of whole cell extracts from heated cells showed the formation of the heat shock transcription factor (HSF)-heat shock element (HSE) complex. Dissociation of HSF from the HSE of the human HSP70 promotor occurred within 4 h under both pH conditions. The kinetics of recovery were not affected by treatment with 0.1% dimethyl sulfoxide (DMSO). However, the dissociation of HSF-HSE complex was markedly delayed during treatment with a combination of low pH and QCT. In addition,in vitro transcription assays showed a suppression of initiation and elongation of HSP70 mRNA. These results may explain why the combination of low pH and QCT treatment suppresses and delays the HSP70 gene expression as well as thermotolerance development.

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References

  1. Tissieres A, Mitchell HK, Tracy U: Protein synthesis in salivary glands ofDrosophila melanogaster. Relation to chromosome puffs. J Mol Biol 84: 389–398, 1974

    Google Scholar 

  2. Peluso RW, Lamb RA, Choppin PW: Polypeptide synthesis in simian virus 5-infected cells. J Viron 23: 177–187, 1977

    Google Scholar 

  3. Li GC: Induction of thermotolerance and enhanced heat shock protein synthesis in Chinese hamster fibroblasts by sodium arsenite and by ethanol. J Cell Physiol 115: 116–122, 1983

    Google Scholar 

  4. Li GC, Werb Z: Correlation between synthesis of heat shock protein synthesis in Chinese hamster fibroblasts. Proc Natl Acad Sci USA 79: 3218–3222, 1982

    Google Scholar 

  5. Landry J, Bernier D, Chretien P, Nicole LM, Tanguay RM, Marceau N: Synthesis and degradation of heat shock proteins during development and decay of thermotolerance. Cancer Res 42: 2457–2461, 1982

    Google Scholar 

  6. Li GC, Mac JY: Induction of heat shock protein synthesis in murine tumors during the development of thermotolerance. Cancer Res 45: 3816–3824, 1985

    Google Scholar 

  7. Li GC: Elevated levels of 70,000 dalton heat shock protein in transiently thermotolerant Chinese hamster fibroblasts and their stable heat resistant variants. Int J Radiat Oncol Biol Phys 11: 165–177, 1985

    Google Scholar 

  8. Hosokawa N, Nirayoshi K, Nakai A, Hosokawa Y, Marui N, Yoshida M, Sakai T, Nishino H, Aoike A, Kawai K, Nagata K: Flavonoids inhibit the expression of heat shock proteins. Cell Struct Funct 15: 383–401, 1990

    Google Scholar 

  9. Kim JH, Kim SH, Alfieri AA, Young CW: Quercetin, an inhibitor of lactate transport and a hyperthermic sensitizer of HeLa cells. Cancer Res 44: 102–106, 1984

    Google Scholar 

  10. Lee YJ, Hou Z, Curetty L, Borrelli MJ, Corry PM: Correlation between redistribution of a 26 kDa protein and development of chronic thermotolerance in various mammalian cell lines. J Cell Physiol 145: 324–332, 1990

    Google Scholar 

  11. Levy J, Teuerstein I, Marbach M, Radian S, Sharoni Y: Tyrosine protein kinase activity in the DMBA-induced rat mammary tumor: inhibition by quercetin. Biochem Biophys Res Commun 123: 1227–1233, 1984

    Google Scholar 

  12. Cochet C, Feige JJ, Pirollet F, Keramidas M, Chambaz EM: Selective inhibition of a cyclic nucleotide independent protein kinase (G type casein kinase) by quercetin and related polyphenols. Biochem Pharmacol 31: 1357–1361, 1982

    Google Scholar 

  13. Gschwendt M, Horn F, Kittstein W, Marks F: Inhibition of the calcium-and phospholipid-dependent protein kinase activity from mouse brain cytosol by quercetin. Biochem Biophys Res Commun 117: 444–447, 1983

    Google Scholar 

  14. Srivastava AK: Inhibition of phosphorylase kinase, and tyrosine protein kinase activities by quercetin. Biochem Biophys Res Commun 131: 1–5, 1985

    Google Scholar 

  15. Matter WF, Brown RF, Vlahos CJ: The inhibition of phosphatidylinositol 3-kinase by quercetin and analogs. Biochem Biophys Res Commun 186: 624–631, 1992

    Google Scholar 

  16. Belt JA, Thomas JA, Buchsbaum RN, Racker E: Inhibition of lactate transport and glycolysis in Ehrlich ascites tumor cells by bioflavonoids. Biochemistry 18: 3506–3511, 1979

    Google Scholar 

  17. Lee YJ, Curetty L, Hou Z, Kim SH, Kim JH, Corry PM: Effect of pH on quercetin-induced suppression of heat shock gene expression and thermotolerance development in HT-29 cells. Biochem Biophys Res Commun 186: 1121–1128, 1992

    Google Scholar 

  18. Highfield DP, Holahan EV, Holahan PK, Dewey WC: Hyperthermic survival of Chinese hamster ovary cells as a function of cellular population density at the time of plating. Radiat Res 97: 139–153, 1984

    Google Scholar 

  19. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  20. Mosser DD, Theodorakis NG, Morimoto RI: Coordinate changes in heat shock element binding activity and hsp 70 gene transcription rates in human cells. Mol Cell Biol 8: 4736–4744, 1988

    Google Scholar 

  21. Hunt C, Morimoto RI: Conserved features of eukaryotic hsp 70 genes revealed by comparison with the nucleotide sequence of human hsp 70. Proc Natl Acad Sci USA 82: 6455–6459, 1985

    Google Scholar 

  22. Banerji SS, Theodorakis NG, Morimoto RI: Heat shock-induced translational control of HSP70 and globin synthesis in chicken reticulocytes. Mol Cell Biol 4: 2437–2448, 1984

    Google Scholar 

  23. Wu B, Hunt C, and Morimoto R: Structure and expression of the human gene encoding major heat shock protein HSP70. Mol Cell Biol 5: 330–341, 1985

    Google Scholar 

  24. Wilson GN, Hollar BA, Waterson JR, Schmickel RD: Molecular analysis of cloned human 18S ribosomal DNA segments. Proc Natl Acad Sci USA 75: 5367–5371, 1978

    Google Scholar 

  25. Tushinski R, Sussman P, Yu L, Bancroft F: Pregrowth hormone messenger RNA: Glucocorticoid induction and identification in rat pituitary cells. Proc Natl Acad Sci USA 74: 2357–2361, 1977

    Google Scholar 

  26. Lebrach H, Diamond L, Wozney J, Boedtker H: RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry 16: 4743–4751, 1977

    Google Scholar 

  27. Koishi M, Hosokawa N, Sato M, Nakai A, Hirayoshi K, Hiraoka M, Abe M, Nagata K: Quercetin, an inhibitor of heat shock protein synthesis, inhibits the acquisition of thermotolerance in a human colon carcinoma cell line. Jpn J Cancer Res 83: 1216–1222, 1992

    Google Scholar 

  28. Peterson R, Lindquist S: The Drosophila hsp70 message is rapidly degraded at nomral temperature and stabilized by heat shock. Gene 72: 161–168, 1988

    Google Scholar 

  29. Theodorakis N, Morimoto R: Posttranscriptional regulation of hsp70 expression in human cells: effects of heat shock, inhibition of protein synthesis, and adenovirus infection on translation and mRNA stability. Mol Cell Biol 7: 4357–4368, 1987

    Google Scholar 

  30. Pelham HRB: A regulatory upstream promoter element in theDrosophila hsp70 heat-shock gene. Cell 30: 517–528, 1982

    Google Scholar 

  31. Sorger PK, Lewis MJ, Pelham HRB: Heat shock factor is regulated differently in yeast and HeLa cells. Nature (Lond.) 329: 81–84, 1987

    Google Scholar 

  32. Kingston RE, Schuetz TJ, Larin Z: Heat-inducible human factor that binds to a human hsp70 promoter. Mol Cell Biol 7: 1530–1534, 1987

    Google Scholar 

  33. Sorger PK, Pelham HRB: Yeast heat shock factor is an essential DNA-binding protein that exhibits temperature-dependent phosphorylation. Cell 54: 855–864, 1988

    Google Scholar 

  34. Zimarino V, Wu C: Induction of sequence-specific binding ofDrosophila heat shock activator protein without protein synthesis. Nature (Lond.) 327: 727–730, 1987

    Google Scholar 

  35. Abravaya K, Phillips B, Morimoto RI: Attenuation of the heat shock response in HeLa cells is mediated by the relaease of bound heat shock transcription factor and is modulated by changes in growth and in heat shock temperatures. Genes and Dev 5: 2117–2127, 1991

    Google Scholar 

  36. Morimoto RI, Sarge KD, Abravaya K: Transcriptional regulation of heat shock genes. J Biol Chem 267: 21987–21990, 1992

    Google Scholar 

  37. Sorger PK, Nelson HCM: Trimerization of a yeast transcriptional activator via a coiled-coil motif. Cell 59: 807–813, 1989

    Google Scholar 

  38. Cunniff NF, Wagner J, Morgan WD: Modular recognition of 5-basepair DNA sequence motifs by human heat shock transcription factor. Mol Cell Biol 11: 3504–3514, 1991

    Google Scholar 

  39. Rabindran SK, Haroun RI, Clos J, Wisniewski J, Wu C: Regulation of heat shock factor trimer formation: role of a conserved leucine zipper. Science 259: 230–234, 1993

    Google Scholar 

  40. Westwood JT, Clos J, Wu C: Stress-induced oligomerization and chromosomal relocalization of heat-shock factor. Nature 353: 822–827, 1991

    Google Scholar 

  41. Bruce JL, Price BD, Coleman CN, Calderwood SK: Oxidative injury rapidly activates the heat shock transcription factor but fails to increase levels of heat shock proteins. Cancer Res 53: 12–15, 1993

    Google Scholar 

  42. Sarge KD, Murphy SP, Morimoto RI: Activation of heat shock gene transcription by heat shock factor 1 involves oligomerization, acquisition of DAN-binding activity, and nuclear localization and can occur in the absence of stress. Mol Cell Biol 13: 1392–1407, 1993

    Google Scholar 

  43. Clos J, Westwood JT, Becker PB, Wilson S, Lambert K, and Wu C: Molecular cloning and expression of a hexameric Drosophila heat shock factor subject to negative regulation. Cell 63: 1085–1097, 1990

    Google Scholar 

  44. Hosakawa N, Hirayoshi K, Kudo H, Takechi H, Aoike A, Kawai K, Nagata K: Inhibition of the activation of heat shock factor in vivo and in vitro by flavonoids. Mol Cell Biol 12: 3490–3498, 1992

    Google Scholar 

  45. Mosser DD, Duchaine J, Massie B: The DNA-binding activity of the human heat shock transcription factor is regulated in vivo by hsp70. Mol Cell Biol 13: 54275438, 1993

    Google Scholar 

  46. Price B, Calderwood SK: Heat-induced transcription from RNA polymerases II and III and HSF binding activity are coordinately regulated by the products of the heat shock genes. J Cell Physiol 153: 392–401, 1992

    Google Scholar 

  47. Hensold JO, Hunt CR, Calderwood SK, Housman DE, Kingston RE: DNA binding of heat shock factor to the heat shock element is insufficient for transcriptional activation in murine erythroleukemia cells. Mol Cell Biol 10: 1600–1608, 1990

    Google Scholar 

  48. Jurivich DA, Sistonen L, Kroes RA, Morimoto RI: Effect of sodium salicylate on the human heat shock response. Science 255: 1243–1245, 1992

    Google Scholar 

  49. Larson JS, Shuetz TJ, Kingston RE: Activationin vitro of sequence-specific DNA binding by a human regulatory factor. Nature 335: 372–275, 1988

    Google Scholar 

  50. Jakobsen BK, Pelham HRB: Constitutive binding of yeast heat-shock factor to DNAin vivo. Mol Cell Biol 8: 5040–5042, 1988

    Google Scholar 

  51. Nieto-Sotelo J, Wiederrecht G, Okuda A, Parker CS: The yeast heat shock transcription factor contains a transcriptional activation domain whose activity is repressed under nonshock conditions. Cell 62: 807–817, 1990

    Google Scholar 

  52. Chang NT, Huang LE, Liu AYC: Okadaic acid markedly potentiates the heat-induced hsp 70 promoter activity. J Biol Chem 268: 1436–1439

  53. Murai T, Hahn GM, Mivechi NF: Regulation of heat shock transcription factor by ser/thr and tyr-phosphatases.In: Thirteen Annual Meeting of the North American Hyperthermia Society, Abstract P-14-18, 1993

  54. Wooten MW: Alterations in protein kinase C type III-α during heat shock of rat embryo fibroblasts. Exp Cell Res 193: 274–278, 1991

    Google Scholar 

  55. Auger EA, Giaccia AJ, Brown JM, Hahn GM: Evidence that protein kinase C is involved in the activation of heat shock transcription factor by ionizing radiation. In: E. Gerner (ed.) Hyperthermic Oncology, Vol I, Tucson, Arizona, 1992, p99

  56. Kantengwa S, Polla BS: Flavonoids, but not protein kinase C inhibitors, prevent stress protein synthesis during erythrophagocytosis. Biochem Biophys Res Commun 180: 308–314, 1991

    Google Scholar 

  57. Kim SH, Kim JH, Erdos G, Lee YJ: Effect of staurosporine on suppression of heat shock gene expression and thermotolerance development in HT-29 cells. Biochem Biophys Res Commun 193: 759–763, 1993

    Google Scholar 

  58. Choi HS, Li B, Lin Z, Huang E, Liu AY: cAMP and cAMP-dependent protein kinase regulate the human heat shock protein 70 gene promoter activity. J Biol Chem 266: 11858–11865, 1991

    Google Scholar 

  59. Nose K: Inhibition by flavonoids of RNA synthesis in permeable WI-38 cells and of transcription by RNA polymerase II. Biochem Pharmacol 33: 3823–3827, 1984

    Google Scholar 

  60. Rougvie AE, Lis JT: Postinitiation transcriptional control inDrosophila melanogaster. Mol Cell Biol 10: 6041–6045, 1990

    Google Scholar 

  61. O'Brien T, Lis JT: RNA polymerase II pauses at the 5′ end of the transcriptionally inducedDrosophila hsp 70 gene. Mol Cell Biol 11: 5285–5290, 1991

    Google Scholar 

  62. Rougvie AE, Lis JT: The RNA polymerase II molecule at the 5′ end of the uninduced hsp 70 gene ofD. melanogaster is transcriptionally engaged. Cell 54: 795–804, 1988

    Google Scholar 

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Authors and Affiliations

  1. Department of Radiation Oncology, Research Laboratories, William Beaumont Hospital, 3601 West Thirteen Mile Road, 48073, Royal Oak, Michigan, USA

    Yong J. Lee, Geza Erdos, Zi-zheng Hou & Peter M. Corry

  2. Department of Radiation Oncology, Henry Ford Hospital, 2799 West Grand Boulevard, 48202, Detroit, Michigan, USA

    Sang H. Kim & Jae H. Kim

  3. Lucky Biotech Corporation, 5919 Hollis Street, 94608, Emeryville, CA, USA

    Joong M. Cho

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  1. Yong J. Lee
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  4. Sang H. Kim
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  7. Peter M. Corry
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Lee, Y.J., Erdos, G., Hou, Zz. et al. Mechanism of Quercetin-induced suppression and delay of heat shock gene expression and thermotolerance development in HT-29 cells. Mol Cell Biochem 137, 141–154 (1994). https://doi.org/10.1007/BF00944076

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  • DOI: https://doi.org/10.1007/BF00944076

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