Heat Shock Protein 40 (Hsp40) and Hsp70 Protein Expression in Oral Squamous Cell Carcinoma (OSCC)

DOI: 10.4236/jct.2013.43090   PDF   HTML   XML   3,534 Downloads   5,867 Views   Citations


Introduction: As a chaperone, heat shock protein acts as central integrators of protein homeostasis in cell. The form of these functions is to help setting up a complex protein molecular fold (folded protein) in many important settings, such as growth, differentiation, and the ability to live. It has become clear that the control system plays an important role if the folding process fails or an error occurs, causing folding abnormalities and targeted functionality to accumulate. The accumulation of faulty protein folding would harm cells and can result in death. Apparently, there is a correlation between protein folding error with various diseases, such as diabetes mellitus and cancer. Method: We examined protein levels in all samples using Dotblott with monoclonal antibody anti-Hsp40 and anti-Hsp70. Levels of the protein content was read using a densitometer. Modification of Dot Blot was as follows: treatment was conducted with 3 × SSC, added with 20 mL blocking solution, add with total protein samples of 10 mg/ml on nitrocellulose paper, prehybridized, incubated at 70° for 30 seconds, incubated at 70° for 30 seconds with primary antibody anti-Hsp40 or Hsp70 protein and then added with second antibody HRP anti-Hsp40 or Hsp70 protein, treated with 3 × SSC and visualized with TSA HRP, and then administered with streptavidin, biothynil tyramide, and, finally, added with chromogen (DAB) in a confined space. Result: From the analysis of the data using Manova test with Wilks Lambda, there were significant differences in the levels of Hsp40 between Benign Oral Lesion (mean 688.31 area) and OSCC (mean 1354.59 area) patients (p < 0.070), there was also a highly significant difference in Hsp70 levels between patients who experienced Benign Oral Lesion (mean 529.82 area) and OSCC (mean 1346.32 area) patients (p < 0.006). Conclusion: OSCC patients have increased Hsp70 levels, so it is possible that something is going wrong in protein folding. Errors in protein folding result in a new homeostasis or inhibition of apoptosis and increasing cell proliferation that triggers carcinogenesis. Hsp40 acts as co-chaperones.

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A. Prayitno, E. Asnar, O. Astirin, D. Rosmala and S. Putra, "Heat Shock Protein 40 (Hsp40) and Hsp70 Protein Expression in Oral Squamous Cell Carcinoma (OSCC)," Journal of Cancer Therapy, Vol. 4 No. 3, 2013, pp. 734-741. doi: 10.4236/jct.2013.43090.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] M. P. Mayer and B. Bukau, “Hsp70 Chaperones: Cellular Functions and Molecular Mechanism,” Cellular and Molecular Life Sciences, Vol. 62, No. 6, 2005, pp. 670-684. doi:10.1007/s00018-004-4464-6
[2] J. P. Hendrick and F. U. Hartl, “The Role of Molecular Chaperones in Protein Folding,” FASEB Journal, Vol. 9, No. 15, 1995, pp. 1559-1569.
[3] M. P. Goetz, D. O. Toft, M. M. Ames and C. Erlichman, “The Hsp90 Chaperone Complex as a Novel Target for Cancer Therapy,” Annals of Oncology, Vol. 14, No. 8, 2003, pp. 1169-1176. doi:10.1093/annonc/mdg316
[4] C. Spiess, A. S. Meyer, S. Reissmann and J. Frydman, “Mechanism of the Eukaryotic Chaperonin: Protein Folding in the Chamber of Secrets,” Trends in Cell Biology, Vol. 14, No. 11, 2004, pp. 598-604. doi:10.1016/j.tcb.2004.09.015
[5] R. J. Nelson, T. Ziegelhoffer, C. Nicolet, M. WernerWashburne and E. A. Craig, “The Translation Machinery and 70 kd Heat Shock Protein Cooperate in Protein Synthesis,” Cell, Vol. 71, No. 1, 1992, pp. 97-105. doi:10.1016/0092-8674(92)90269-I
[6] J. Frydman, E. Nimmesgern, K. Ohtsuka and F. U. Hartl, “Folding of Nascent Polypeptide Chains in a High Molecular Mass Assembly with Molecular Chaperones,” Nature, Vol. 370, 1994, pp. 111-117. doi:10.1038/370111a0
[7] R. J. Schumacher, R. Hurst, W. P. Sullivan, N. J. Mc Mahon, D. O. Toft and R. L. Matts, “ATP-Dependent Chaperoning Activity of Reticulocyte Lysate,” The Journal of Biological Chemistry, Vol. 269, No. 13, 1994, pp. 9493-9499.
[8] J. P. Hendrick and F. U. Hartl, “The Role of Molecular Chaperones in Protein Folding,” FASEB Journal, Vol. 9, No. 15, 1995, pp. 1559-1569.
[9] J. P. Hendrick, T. Langer, T. A. Davis, F. U. Hartl and M. Wiedmann, “Control of Folding and Membrane Translocation by Binding of the Chaperone DnaJ to Nascent Polypeptides,” Proceedings of the Natinal Academy of Sciences of the USA, Vol. 90, No. 21, pp. 10216-10220. doi:10.1073/pnas.90.21.10216
[10] A. Smith, “Protein Misfolding,” Nature Reviews Drug Discovery, Vol. 426, No. 6968, 2003, pp. 883-909.
[11] A. H. Lund and M. van Lohuizen, “Epigenetics and Cancer,” Genes & Development, Vol. 18, No. 19, 2004, p. 231535.
[12] T. J. Hubbard and C. Sander, “The Role of Heat-Shock and Chaperone Proteins in Protein Folding: Possible Molecular Mechanisms,” Protein Engineering, Vol. 4, No. 7, 1991, pp. 711-717.
[13] H. M. Beere, B. B. Wolf, K. Cain, D. D. Mosser, A. Mahboubi, T. Kuwana, P. Tailor, R. I. Morimoto, G. M. Cohen and D. R. Green, “Heat-Shock Protein 70 Inhibits Apoptosis by Preventing Recruitment of Procaspase-9 to the Apaf1apoptosome,” Nature Cell Biology, Vol. 2, 2000, pp. 469-475. doi:10.1038/35019501
[14] H. M. Beere, “The Stress of Dying: The Role of Heat Shock Proteins in the Regulation of Apoptosis,” Journal of Cell Science, Vol. 117, No. 13, 2004, pp. 2641-2651. doi:10.1242/jcs.01284
[15] S. Arawaka, Y. Machiya and T. Kato, “Heat Shock Proteins as Suppressors of Accumulation of Toxic Prefibrillar Intermediates and Misfolded Proteins in Neurodegenerative Diseases,” Current Pharmaceutical Biotechnology, Vol. 11, No. 2, 2010, pp. 158-166.
[16] S. A. Houck, S. Singh and D. M. Cyr, “Cellular Responses to Misfolded Proteins and Protein Aggregates,” Methods in Molecular Biology, Vol. 832, 2012, pp. 455-461. doi:10.1007/978-1-61779-474-2_32
[17] J. L. Alberto, M. D. Macario and E. C. de Macario, “Sick Chaperones, Cellular Stress, and Disease,” The New England Journal of Medicine, Vol. 353, No. 14, 2005, pp. 1489-1501.
[18] M. M. M. Wilhelmus, R. M. W. de Waal and M. M. Verbeek, “Heat Shock Proteins and Amateur Chaperones in Amyloid-Beta Accumulation and Clearance in Alzheimer’s Disease,” Molecular Neurobiology, Vol. 35, No. 3, 2007, pp. 203-216.
[19] H. L. Schmits, “Species Diagnostics Protocols: PCR and Other Nucleic Acid Methods,” In: J. P. C. Humana, Methods in Molecular Biology, Press Inc., Totowa, 1994.
[20] T. Hessel, S. P. Dhital, R. Plank and D. Dean, “Immune Response to Chlamydial 60-Kilodalton Heat Shock Protein in Tears from Nepali Trachoma Patients,” Infection and Immunity, Vol. 69, No. 8, 2001, pp. 4996-5000.
[21] M. F. Prummel, Y. Van Pareren, O. Barker and W. M. Wiersinga, “Anti-Heat Shock Protein (hsp)72 Antibodies Are Present in Patients with Graves’ Disease (GD) and in Smoking Control Subjects,” Clinical and Experimental Immunology, Vol. 110, No. 2, 1997, pp. 292-295.
[22] C. Georgopoulos and W. J. Welch, “Role of the Major Heat Shock Proteins as Molecular Chaperones,” Annual Review of Cell Biology, Vol. 9, 1993, pp. 601-634. doi:10.1146/annurev.cb.09.110193.003125
[23] K. N. Truscott, K. Brandner and N. Pfanner, “Mechanism of Protein Import into Mitochondria,” Current Biology, Vol. 13, No. 8, 2003, pp. R326-R337. doi:10.1016/S0960-9822(03)00239-2
[24] S. Alberti, C. Esser and J. Hohfeld, “BAG-1—A Nucleotide Exchange factor Of Hsc70 with Multiple Cellular Functions,” Cell Stress and Chaperones, Vol. 8, No. 3, 2003, pp. 225-231. doi:10.1379/1466-1268(2003)008<0225:BNEFOH>2.0.CO;2
[25] A. J. Caplan, “What is a Co-Chaperone?” Cell Stress and Chaperones, Vol. 8, No. 2, 2003, pp. 105-107. doi:10.1379/1466-1268(2003)008<0105:WIAC>2.0.CO;2
[26] J. C. Young, J. M. Barral and F. U. Hartl, “More than Folding: Localized Functions of Cytosolic Chaperones,” Trends in Biochemical Sciences, Vol. 28, No. 10, 2003, pp. 541-547. doi:10.1016/j.tibs.2003.08.009
[27] J. C. Young, V. R. Agashe, K. Siegers and F. U. Hartl, “Pathways of Chaperone-Mediated Protein Folding in the Cytosol,” Nature Reviews Molecular Cell Biology, Vol. 5, No. 10, 2004, pp. 781-791. doi:10.1038/nrm1492
[28] J. Martin, “Chaperonin Function—Effects of Crowding and Confinement,” Journal of Molecular Recognition, Vol. 17, No. 5, 2004, pp. 465-472. doi:10.1002/jmr.707
[29] O. O. Odunuga, V. M. Longshaw and G. L. Blatch, “Hop: More than an HSP70/Hsp90 Adaptor Protein,” BioEssays, Vol. 26, No. 10, 2004, pp. 1058-1068. doi:10.1002/bies.20107
[30] A. L. Fink, “Chaperone-Mediated Protein Folding,” Physiological Reviews, Vol. 79, No. 2, 1999, pp. 425-449.
[31] B. Kleizen and I. Braakman, “Protein Folding and Quality Control in the Endoplasmic Reticulum,” Current Opinion in Cell Biology, Vol. 16, No. 4, 2004, pp. 343-349. doi:10.1016/j.ceb.2004.06.012
[32] A. J. L. Macario and E. C. de Macario, “The Pathology of Cellular Anti-Stress Mechanisms: A New Frontier,” Stress, Vol. 7, No. 4, 2004, pp. 243-249. doi:10.1080/10253890400019706
[33] A. J. L. Macario and E. C. de Macario, “Molecular Chaperones and Age-Related Degenerative Disorders,” Advances in Cell Aging and Gerontology, Vol. 7, 2001, pp. 131-162. doi:10.1016/S1566-3124(01)07018-3
[34] G. A. Gaitanaris, A. Vysokanov, S. C. Hung, M. E. Gottesman and A. Gragerov, “Successive Action of Escherichia coli Chaperones in Vivo,” Molecular Microbiology, Vol. 14, No. 5, 1994, pp. 861-869. doi:10.1111/j.1365-2958.1994.tb01322.x
[35] M. A. Petit, W. Bedale, J. Osipiuk, C. Lu, M. Rajagopalan, P. Mc Inerney, M. F. Goodman and H. Echols, Sequential Folding of UmuC by the Hsp70 and Hsp60 Chaperone Complexes of Escherichia coli,” The Journal of Biological Chemistry, Vol. 269, No. 38, 1994, pp. 23824-23829.
[36] A. A. Morino and D. H. Moris, “Chronic Electromagnetic Stressors in the Enviroment: A Risk Factor in Human Cancer,” Journal of Environmental Science and Health, Vol. 3, No. 2, 1985, pp. 189-219.
[37] P. L. Ooi and K. T. Goh, “Sick Building Syndrome: An Emerging Stress-Related Disorders,” International Journal of Epidemiology, Vol. 26, No. 6, 1997, pp. 1243-1249.
[38] K. Sjovall, B. Attner, T. Lithman, D. Noreen, B. Gunnars, B. Thomé, L. Lidgren, H. Olsson and M. Englund, “Sick Leave of Spouses to Cancer Patients before and after Diagnosis,” Acta Oncologica, Vol. 49, No. 4, 2010, pp. 467-473.
[39] P. Leandro and C. M. Gomes, “Protein Misfolding in Conformational Disorders: Rescue of Folding Defects and Chemical Chaperoning,” Mini-Reviews in Medicinal Chemistry, Vol. 8, No. 9, 2008, pp. 901-911.
[40] A. Prayitno, “Cervical Cancer with Human Papilloma virus and Epstein Barr virus Positif,” Journal of Carcinogenesis, Vol. 5, 2006, p.13.
[41] A. K. Chaturvedi, “Beyond Cervical Cancer: Burden of Other HPV-Related Cancers among Men and Women,” The Journal of Adolescent Health, Vol. 46, No. 4S, 2010, pp. S20-S26. doi:10.1016/j.jadohealth.2010.01.016
[42] A. Prayitno, “Incidence of HPV Infection in Oral Squamous Cell Carcinoma Its Associated with the Presence of p53 and C-Myc Mutation : A Case Control Study in Muwardi Hospital Surakarta,” Indian Journal of Dental Research, Vol. 17, No. 2, 2010, pp. 48-52.
[43] D. R. Ciocca and S. K. Calderwood, “Heat Shock Proteins in Cancer: Diagnostic, Prognostic, Predictive, and Treatment Implications,” Cell Stress Chaperones, Vol. 10, No. 2, 2005, pp. 86-103.
[44] J. Klucken, Y. Shin, E. Masliah, B. T. Hyman and P. J. McLean, “HSP70 Reduces Alphasynuclein Aggregation and Toxicity,” The Journal of Biological Chemistry, Vol. 279, 2004, pp. 25497-25502. doi:10.1074/jbc.M400255200
[45] J. Magrane, R. C. Smith, K. Walsh and H. W. Querfurth, “Heat Shock Protein 70 Participates in the Neuroprotective Response to Intracellularly Expressed Beta-Amyloid in Neurons,” The Journal of Neuroscience, Vol. 24, No. 7, 2004, pp. 1700-1706. doi:10.1523/JNEUROSCI.4330-03.2004
[46] C. A. Ross and M. A. Poirier, “Protein Aggregation and Neurodegenerative Disease,” Nature Medicine, Vol. 10, 2004, pp. S10-S17. doi:10.1038/nm1066
[47] M. Arrasate, S. Mitra, E. S. Schweitzer, M. R. Segal and S. Finkbeiner, “Inclusion Body Formation Reduces Levels of Mutant Huntingtin and the Risk of Neuronal Death,” Nature, Vol. 431, 2004, pp. 805-810. doi:10.1038/nature02998
[48] M. A. Poirier, H. Jiang and C. A. Ross, “A StructureBased Analysis of Huntingtin Mutant Polyglutamine Aggregation and Toxicity: Evidence for a Compact BetaSheet Structure,” Human Molecular Genetics, Vol. 14, No. 6, 2005, pp. 765-774. doi:10.1093/hmg/ddi071
[49] A. J. L. Macario and E. C. de Macario, “Genetic Disorders Involving Molecularchaperone Genes: A Perspective,” Genetics in Medicine, Vol. 7, 2005, pp. 3-12. doi:10.1097/01.GIM.000015135 1.11876.C3
[50] I. A. Ruz, D. A. Ossa, W. K. Torres, U. Kemmerling, B. A. Rojas and C. A. Martínez, “Nucleolar Organizer Regions in a Chronic Stress and Oral Cancer Model,” Oncology Letters, Vol. 3, No. 3, 2012, pp. 541-544.
[51] C. Bernstein, H. Bernstein, C. M. Payne and H. Garewal, “DNA Repair/Pro-Apoptotic Dual-Role Proteins in Five Major DNA Repair Pathways: Fail-Safe Protection against Carcinogenesis,” Mutation Research, Vol. 511, No. 2, 2002, pp. 145-178.
[52] F. Lang, M. Ritter, N. Gamper, S. Huber, S. Fillon, V. Tanneur, A. Lepple-Wienhues, I. Szabo and E. Gulbins, Cell Volume in the Regulation of Cell Proliferation and Apoptotic Cell Death,” Cell Physiology Biochemistry, Vol. 10, No. 5-6, 2000, pp. 417-428. doi:10.1159/000016367
[53] R. S. Fife, B. T. Rougraff, C. Proctor and G. W. Sledge Jr., “Inhibition of Proliferation and Induction of Apoptosis by Doxycycline in Cultured Human Osteosarcoma Cells,” The Journal of Laboratory and Clinical Medicine, Vol. 130, No. 5, 1997, 530-534.
[54] A. P. Feinberg, R. Ohlsson and S. Henikoff, “The Epigenetic Progenitor Origin of Human Cancer,” Nature Reviews Genetics, Vol. 7, 2006, pp. 21-33.

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