Immunohistochemical Aspects on (HSP70) and its Correlation to Clinical Stages, Pathological Grading and Hormones Receptors (ER, PR, HER-2) in Breast Cancer (BC) Women from South of Iraq

Author(s): Alaa R. Rashed, Maha K. Ibrahim, Maha SH. Hassan

Breast cancer considered the most common malignancy .Therefore this study was aimed to review the observations and information on relationship of preoperative level of (HSP70) with hormones receptors (ER,PR,HER-2) and the expression of these receptors in biopsies from patients with different stages and grades of disease by immunohistochemical method. In this study total number of (130)cases was analyzed and most patients diagnosed with stage (T111) then (T11)and (T1) with highly significant difference at (p≤ 0.05) and the same results was obtained from classified tumor grading with (BC)patients. The recent study evaluated level of (HSP70) in all (BC)patients ,the data showed highly significant difference and the mean level was increased in (71)case compared to control ,also we evaluated the relationship between tumor staging and grading with level of (HSP70) and data revealed to high level in (T111) patients while no signification between (T1,T3) ,the same results recorded between tumor grading and level (HSP70), hormones receptors expression (ER,PR,HER-2) and its correlation with (IL-6)level was also determined ,the( HSP70) with (ER+)compared to (ER-) with high significance (p=0.00),the same results shown with (PR+) and(HER-2) expression and (HSP70)in all (BC)patients. The role of (HSP70) in (BC) immunity ,carcinogenesis ,cancer pathogenesis ,tumor initiation ,growth and metastasis .Results also explained correlate of (HSP70)with clinical staging and grading ,these findings pointed to an important role in the progression of (BC) at least in patients with advanced disease stages. Data determined that the (ER+)(PR+) positive tumor more reactive with(HSP70) which explained the effect of this factor on hormones receptors. Moreover present study showed little elevated in concentrate of (HSP70) with Her2+as compared with Her2- but with no significant differences,(p=.158).Immunhistochemical staining was used in this study and results showed variable degree of staining Within invasive ductal and lobular carcinoma types compared to control.

1. Kolahdoozan, S., Sadjadi, A., Radmard, A.R., & Khademi, H. (2010). Five common cancers in Iran. Archives of Iranian Medicine, 13(2), 143–6.
2. Balaburski, G. M, Ju Leu J., Beeharry N., Hayik S., . Andrake M. D, Zhang G , Herlyn M., . Villanueva J,1 Dunbrack, Yen T., George D. L., and Murphy1, M. E. ( 2013). A modified HSP70 inhibitor shows broad activity as an anticancer agent . Mol Cancer Res . 11(3): 219–229.
3. Ciocca DR, Calderwood SK (2005) Heat shock proteins in cancer: diagnostic, prognostic, predictive, and treatment implications. Cell Stress Chaperones 10(2):86–103 .
4. Daugaard M, Rohde M, and Jäättelä M. (2007).The heat shock protein 70 family: Highly homologous proteins with overlapping and distinct functions. FEBS Lett.581:3702-10.
5. Ferlay J., Shin H. R., Bray F., Forman D., Mathers C., and. Parkin D. M.(2010).“Estimates of worldwide burden of cancer in 2008: GLOBOCAN 2008,” International Journal of Cancer, 127( 12) : 2893–2917.
6. Garrido C., Gurbuxani S, Ravagnan L., and Kroemer G.( 2001). Heat shock proteins: endogenous modulators of apoptotic cell death. Biochem Biophys Res Commun 286: 433-442.
7. Granato M , LacconV i, Peddis M , Lotti L V , Renzo L D , Gonnella R, Santarelli R, Trivedi P, Frati L,’Orazi G D, Faggioni A and Cirone M . (2013). HSP70 inhibition by 2-phenylethynesulfonamide induces lysosomal cathepsin D release and immunogenic cell death in primary effusion lymphoma Open. Cell Death and Disease (4):730.
8. Margel D. , Pesvner-Fischer, M. , Baniel J., Yossepowitch O., and, Cohen I. R. ( 2 0 1 1 ) . Stress Proteins and Cytokines are Urinary Biomarkers for Diagnosis and Staging of Bladder Cancer. European Urology. (59):113-115.
9. Powers, M.V., Jones, K., Barillari C. , West wood I. , van Montfort R.L., and Workman P. ( 2010) .Targeting HSP70: the second potentially druggable heat shock protein and molecular chaperone? Cell Cycle;9:1542-50.
10. Rohde M, Daugaard M, Jensen MH, Helin K, Nylandsted J, and Ja¨a¨ttela¨ M. (2005).Members of the heat-shock protein 70 family promote cancer cell growth by distinct mechanisms. Genes Dev;19:570–82.
11. Sherman M., and Multhoff G. (2007) Heat shock proteins in cancer. Ann N Y Acad Sci., 1113, 192-201.
12. Vargas-Roig L M ,Gago F E, Tello O, Aznar J C, and Ciocca D R (1998). Int J Cancer 73:468–475
13. Dundas SR, Lawrie LC, Rooney PH, Murray GI. 2005. Mortalin is over-expressed by colorectal adenocarcinomas and correlates with poor survival. J Pathol 205: 74–81
14. Watson RWG, Lebret T, Fitzpatrick JM. 2003. Heat shock proteins in the genitourinary system. Curr Urol Rep 4: 70–76.
15. Suzuki,K.,Ito,Y.,Wakai,K.,Kawado, M., Hashimoto,S.,Seki,N.,Ando, M., Nishino,Y.,Kondo,T.,Watan- abe, Y.,Ozasa,K.,Inoue,T.,and Tamakoshi,A.(2006).Serumheat shock protein70levelsandlung cancerrisk:acase-controlstudy nestedinalargecohortstudy. Can-cer Epidemiol.BiomarkersPrev. 15, 1733–1737.
16. Didelot, C.,Lanneau,D.,Brunet,M., Joly,A.L.,DeThonel,A.,Chiosis,G., and Garrido,C.(2007).Anti-cancer therapeuticapproachesbasedon intracellularandextracellularheat shock proteins. Curr.Med.Chem. 14, 2839–2847.
17. Tavassol F, Starke OF, Kokemuller H, Wegener G, Muller-Tavassol CC, Gellrich NC, et al. Prognostic significance of heat shock protein 70 (HSP70) in patients with oral cancer. Head Neck Oncol. 2011; 3:10. [PubMed: 21345207]
18. Laoukili J, Stahl M, Medema RH (2007) FoxM1: At the crossroads of ageing and cancer. Biochim Biophys Acta 1775: 92-102.
19. Lebret, T., Watson, R. W. G., Molinié, V., O'Neill, A., Gabriel, C., Fitzpatrick, J. M. and Botto, H. (2003), Heat shock proteins HSP27, HSP60, HSP70, and HSP90. Cancer, 98: 970–977. doi:10.1002/cncr.11594
20. Halasi M, Váraljai R , Benevolenskaya E and Andrei L. Gartel.(2015), Novel function of molecular chaperone HSP70: suppression of oncogenic FOXM1 after proteotoxic stress,J. Biol. Chem. published online November 11, 2015 as Manuscript M115.678227
21. Rohde M, Daugaard M, Jensen MH, Helin K, Nylandsted J, et al. (2005) Members of the heat-shock protein 70 family promote cancer cell growth by distinct mechanisms. Genes Dev 19: 570-582.
22. Wadhwa R, Takano S, Kaur K, Deocaris CC, Pereira-Smith OM, Reddel RR, et al. Upregulation of mortalin/mthsp70/Grp75contributes to human carcinogenesis. Int J Cancer. 2006;118:2973–80.
23. Yang L, Liu X, Hao J, Yang Y, Zhao M, Zuo J, et al. Glucose-regulated protein75 suppresses apoptosis induced by glucose deprivation in PC12 cells through inhibition of Bax conformational change. Acta Biochem Biophys Sin (Shanghai). 2008;40(4):339–48.
24. Deocaris CC, Lu WJ, Kaul SC, Wadhwa R. Druggability of mortalin for cancer and neuro-degenerativedisorders. Curr Pharm Des. 2013;19:418–29.
25. Yang L, Guo W, Zhang Q, Li H, Liu X, Yang Y, et al. Crosstalk between Raf/ MEK/ERK and PI3K/AKT insuppression of Bax conformational change by Grp75 under glucose deprivation conditions. J Mol Biol. 2011;414:654–66.
26. Chen J, Liu WB, Jia WD, Xu GL, Ma JL, Huang M, et al. Overexpression of Mortalin in hepatocellular carcinoma and its relationship with angiogenesis and epithelial to mesenchymal transition. Int J Oncol. 2014;44:247–55.
27. Chen X, Xu B, Li H, Yang L, Zuo J, Liu W, et al. Expression of mortalin detected in human liver cancer by tissue microarrays. Anat Rec. 2011;294:1344–51.
28. Haidan Jin, Meiying Ji, Liyan Chen, Qixiang Liu, Shuanlong Che, Ming Xu and Zhenhua Lin ,2016, The clinicopathological significance of Mortalin overexpression in invasive ductal carcinoma of breast, of Experimental & Clinical Cancer Research (2016) 35:42 doi 10.1186/s13046-016-0316-0.
29. S. E. Conroy and D. S. Latchman, “Do Heat Shock Pro-teins Have a Role in Breast Cancer?” British Journal of Cancer, Vol. 74, No. 5, 1996, pp. 717-721. doi:10.1038/bjc.1996.427
30. R. Kumaraguruparan, D. Karunagaran, C. Balachandran, B. M. Manohar and S. Nagini, “Of Human and Canines: A Comparative Evaluation of Heat Shock and Apoptosis- Associated Proteins in Mammary Tumours,” Clinica Chi- mica Acta, Vol. 365, No. 1-2, 2006, pp. 168-176. doi:10.1016/j.cca.2005.08.018
31. M. Romanucci, A. Marinelli, G. Sarli and L. Della Salda, “Heat Shock Proteins Expression in Canine Malignant Mammary Tumours,” BMC Cancer, Vol. 6, 2006, p. 171. doi:10.1186/1471-2407-6-171
32. M. Romanucci, T. Bastow and L. Della Salda, “Heat Shock Proteins in Animal Neoplasms and Human Tumours—A Comparison,” Cell Stress & Chaperones, Vol. 13, No. 3, 2008, pp. 253-262. doi:10.1007/s12192-008-0030-8
33. C. Torronteguy, A. Frasson, F. Zerwes, E. Winnikov, V. D. da Silva, A. Ménoret and C. Bonorino, “Inducible Heat Shock Protein 70 Expression as a Potential Predic- tive Marker of Metastasis in Breast Tumors,” Cell Stress & Chaperones, Vol. 11, No. 1, 2006, pp. 34-43. doi:10.1379/CSC-159R.1
34. C. Torronteguy, A. Frasson, F. Zerwes, E. Winnikov, V. D. da Silva, A. Ménoret and C. Bonorino, “Inducible Heat Shock Protein 70 Expression as a Potential Predic- tive Marker of Metastasis in Breast Tumors,” Cell Stress & Chaperones, Vol. 11, No. 1, 2006, pp. 34-43. doi:10.1379/CSC-159R.1
35. L. M. Vargas-Roig, F. E. Gago, O. Tello, J. C. Aznar and D. R. Ciocca, “Heat Shock Protein Expression and Drug Resistance in Breast Cancer Patients Treated with Induc- tion Chemotherapy,” International Journal of Cancer, Vol. 79, No. 5, 1998, pp. 468-475. doi:10.1002/(SICI)1097-0215(19981023)79:5<468::AID-IJC4>3.0.CO;2-Z
36. L. Chouchane, S. B. Ahmed, S. Baccouche and S. Re- madi, “Polymorphism in the Tumor Necrosis Factor-Al- pha Promotor Region and in the Heat Shock Protein 70 Genes Associated with Malignant Tumors,” Cancer, Vol. 80, No. 8, 1997, pp. 1489-1496. doi:10.1002/(SICI)1097-0142(19971015)80:8<1489::AID-CNCR17>3.0.CO;2-1
37. S. E. Conroy, P. D. Sasieni, I. Fentiman and D. S. Latch- man, “Autoantibodies to the 90kDa Heat Shock Protein and Poor Survival in Breast Cancer Patients,” European Journal of Cancer, Vol. 34, No. 6, 1998, pp. 942-943.
38. C. Didelot, E. Schmitt, M. Brunet, L. Maingret, A. Par-cellier and C. Garrido, “Heat Shock Proteins: Endogenous Modulators of Apoptotic Cell Death,” Handbook of Ex-perimental Pharmacology, Vol. 172, 2006, pp. 171-198. doi:10.1007/3-540-29717-0_8
39. JKalogeraki A, Giannikaki E, Tzardi M, Kafousi M, Ieromonachou P,Dariviannaki K, Askoxylakis J, Tsiftsis D, Stathopoulos E, Zoras O(2007). Correlation of heat shock protein (HSP70) expression with cell proliferation (MIB1), estrogen receptors (ER) and clinicopathological variables in invasive ductal breast carcinomas.Exp. Clin. Cancer Res. 26: 367-368.
40. Yang, Y.; Rao, R.; Shen, J.; Tang, Y.; Fiskus, W.; Nechtman, J.; Atadja, P.; Bhalla, K. Role of acetylation and extracellular location of heat shock protein 90alpha in tumor cell invasion. Cancer Res. 2008, 68, 4833–4842.
41. Hani Z. Asfour, Mohamed H. AlQahtani, Khaled A. Balto and Sufian M. ElAssouli(2011): Heat shock proteins, estrogen and progesterone receptors expression in breast carcinomas Journal of Medicine and Medical Science Vol. 2(7) pp. 977-984, July 2011.
42. Leonardo Della Salda, Mariarita Romanucci:2012, The Role of Heat Shock Proteins in Mammary Neoplasms: A Brief ReviewJournal of Cancer Therapy, 2012, 3, 755-767.
43. Gunaldi M , Kocoglu H, Okuturlar Y, Gedikbasi A, Karabulut M, Alis H, Hursitoglu M.(2015), Heat shock protein 70 is a useful marker for predicting colorectal cancer, JBUON 2015; 20(6): 1464-1470.
44. Amna N. Jassem, Hind M. Mousa,Munther J. Hussain(2014). Heat Shock Protein70 as Biomarker in Bladder Cancer, Baghdad Science Journal , Vol.11(3)2014.
45. Yu H-J, Chang Y-H, Pan C-C. Prognostic significance of heat shock proteins in urothelial carcinoma of the urinary bladder. Histopathology. 2013; 62:788–798. [PubMed: 23425286]
46. Nishitai G, Matsuoka M. 2008. Differential regulation of HSP70 expression by the JNK kinases SEK1 and MKK7 in mouse embryonic stem cells treated with cadmium. J Cell Biochem 104:1771–1780.
47. Henson DE, Ries L, Freedman LS, Carriaga M. Relationship among outcome, stage of disease, and histologic grade for 22,616 cases of breast cancer. The basis for a prognostic index. Cancer. 1991;68:2142-9.
48. AJCC(Amrican Joint Committee on Cancer) cancer Staging Manual,7thed Edge,SB, Byrd,DR, Compton,CC,etal,Springer- Verlag,New York,2010.p.347 377