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:: Volume 21, Issue 2 (7-2019) ::
J Gorgan Univ Med Sci 2019, 21(2): 120-127 Back to browse issues page
Effect of recombinant Herceptin on the growth of breast cancer cells
Nafiseh Kiarostami1 , Jafar Amani 2, Abbas Ali Imani Fooladi , Ali Mirhosseini3
1- M.Sc in Biology, Department of Biology, Faculty of Basic Science, Damghan Branch, Islamic Azad University, Damghan, Iran
2- Associate Professor, Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran , jafar.amani@gmail.com
3- Assistant professor, Applied Microbiology Research Center, Systems Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran
Abstract:   (6549 Views)
Background and Objective: Breast cancer is one of the most common types of cancer among women. HER-2 molecule as the receptor of tyrosine kinase from the family of epithermal growth factor is a major cause of cancer. The Herceptin protein molecule, which is an anti-HER-2 antibody, can play an important role in the diagnosis and treatment of breast cancer. This study was done to subcloning of Herceptin gene, expression in the prokaryotic system (E.coli) and produce Herceptin recombinant protein for use in the treatment of breast cancer.

Methods: In this descriptive – laboratory study, Herceptin gene from synthesized construct was isolated by enzyme digestion, and then subcloned to the expression vector pET28a. Subcloning of the gene was confirmed using PCR and enzyme digestion. After transferring the vector into E.coli BL21 DE3, expression of the recombinant Herceptin gene was induced by IPTG. The recombinant protein was evaluated by SDS-PAGE and purified with Ni-NTA column chromatography and finally verified by western blotting using anti-histidine antibody. The survival of cells adjacent to recombinant Hercaptin by MTT was investigated.

Results: Following the subcloning of the Herceptin gene, PCR and enzyme digestion, the 741 fragment of the Herceptin gene was confirmed. Confirmation of Herceptin's recombinant protein and its evaluation on SDS-PAGE gel about 27 kDa was done. The recombinant protein was also confirmed with anti-histidine tag. The purified protein adjacent to the SKBR3 cell line was able to block the growth of cancer cells.

Conclusion: Regarding the expersion of HER2 antigen on surface of breast cancer cells, Herceptin can act as antibody blocker and it arrests the growth of breast cancer cells.
Keywords: Breast cancer, Recombinant Herceptin, Her-2 antigen, Cancer cell
Full-Text [PDF 366 kb]   (11867 Downloads)    
Type of Study: Original Articles | Subject: Medical Biotecnology
References
1. Thongsuksai P, Chongsuvivatwong V, Sriplung H. Delay in breast cancer care: a study in Thai women. Med Care. 2000 Jan; 38(1): 108-14.
2. Coleman MP, Quaresma M, Berrino F, Lutz JM, De Angelis R, Capocaccia R, et al. Cancer survival in five continents: a worldwide population-based study (CONCORD). Lancet Oncol. 2008 Aug; 9(8): 730-56. doi: 10.1016/S1470-2045(08)70179-7
3. Cheung KL, Graves CR, Robertson JF. Tumour marker measurements in the diagnosis and monitoring of breast cancer. Cancer Treat Rev. 2000 Apr; 26(2): 91-102. doi: 10.1053/ctrv.1999.0151
4. Hayes DF. Tumor markers for breast cancer. Current utilities and future prospects. Hematol Oncol Clin North Am. 1994 Jun; 8(3): 485-506.
5. Duffy MJ, Shering S, Sherry F, McDermott E, O'Higgins N. CA 15-3: a prognostic marker in breast cancer. Int J Biol Markers. 2000 Oct-Dec; 15(4): 330-33.
6. Ebeling FG, Stieber P, Untch M, Nagel D, Konecny GE, Schmitt UM, et al. Serum CEA and CA 15-3 as prognostic factors in primary breast cancer. Br J Cancer. 2002 Apr; 86(8): 1217-22. doi: 10.1038/sj.bjc.6600248
7. Bartlett JM, Going JJ, Mallon EA, Watters AD, Reeves JR, Stanton P, et al. Evaluating HER2 amplification and overexpression in breast cancer. J Pathol. 2001 Nov; 195(4): 422-28. doi: 10.1002/path.971
8. 8. Ludovini V, Gori S, Colozza M, Pistola L, Rulli E, Floriani I, et al. Evaluation of serum HER2 extracellular domain in early breast cancer patients: correlation with clinicopathological parameters and survival. Ann Oncol. 2008 May; 19(5): 883-90. doi: 10.1093/annonc/mdm585
9. Yakes FM, Chinratanalab W, Ritter CA, King W, Seelig S, Arteaga CL. Herceptin-induced inhibition of phosphatidylinositol-3 kinase and Akt Is required for antibody-mediated effects on p27, cyclin D1, and antitumor action. Cancer Res. 2002 Jul; 62(14): 4132-41.
10. Cooley S, Burns LJ, Repka T, Miller JS. Natural killer cell cytotoxicity of breast cancer targets is enhanced by two distinct mechanisms of antibody-dependent cellular cytotoxicity against LFA-3 and HER2/neu. Exp Hematol. 1999 Oct; 27(10): 1533-41.
11. Valabrega G, Montemurro F, Aglietta M. Trastuzumab: mechanism of action, resistance and future perspectives in HER2-overexpressing breast cancer. Ann Oncol. 2007 Jun; 18(6): 977-84. doi: 10.1093/annonc/mdl475
12. Baselga J, Norton L, Albanell J, Kim YM, Mendelsohn J. Recombinant humanized anti-HER2 antibody (Herceptin) enhances the antitumor activity of paclitaxel and doxorubicin against HER2/neu overexpressing human breast cancer xenografts. Cancer Res. 1998 Jul; 58(13): 2825-31.
13. Goldenberg MM. Trastuzumab, a recombinant DNA-derived humanized monoclonal antibody, a novel agent for the treatment of metastatic breast cancer. Clin Ther. 1999 Feb; 21(2): 309-18. doi: 10.1016/S0149-2918(00)88288-0
14. Tolosa L, Donato MT, Gómez-Lechón MJ. General Cytotoxicity Assessment by Means of the MTT Assay. Methods Mol Biol. 2015; 1250: 333-48. doi: 10.1007/978-1-4939-2074-7_26.
15. Davoli A, Hocevar BA, Brown TL. Progression and treatment of HER2-positive breast cancer. Cancer Chemother Pharmacol. 2010 Mar; 65(4): 611-23. doi: 10.1007/s00280-009-1208-1
16. Bange J, Zwick E, Ullrich A. Molecular targets for breast cancer therapy and prevention. Nat Med. 2001 May; 7(5): 548-52. doi: 10.1038/87872
17. Duffy MJ. Serum tumor markers in breast cancer: are they of clinical value? Clin Chem. 2006 Mar; 52(3): 345-51. doi: 10.1373/clinchem.2005.059832
18. Rilke F, Colnaghi MI, Cascinelli N, Andreola S, Baldini MT, Bufalino R, et al. Prognostic significance of HER-2/neu expression in breast cancer and its relationship to other prognostic factors. Int J Cancer. 1991 Aug; 49(1): 44-49.
19. Ross JS, McKenna BJ. The HER-2/neu oncogene in tumors of the gastrointestinal tract. Cancer Invest. 2001; 19(5): 554-68.
20. Widakowich C, Dinh P, de Azambuja E, Awada A, Piccart-Gebhart M. HER-2 positive breast cancer: what else beyond trastuzumab-based therapy? Anticancer Agents Med Chem. 2008; 8(5): 488-96.doi: 10.2174/187152008784533062
21. Romond EH, Perez EA, Bryant J, Suman VJ, Geyer CE Jr, Davidson NE, et al. Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer. N Engl J Med. 2005 Oct; 353(16): 1673-84. doi: 10.1056/NEJMoa052122
22. Campiglio M, Locatelli A, Olgiati C, Normanno N, Somenzi G, Viganò L, et al. Inhibition of proliferation and induction of apoptosis in breast cancer cells by the epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor ZD1839 ('Iressa') is independent of EGFR expression level. J Cell Physiol. 2004 Feb; 198(2): 259-68. doi: 10.1002/jcp.10411
23. Kazemi M, Amani J, Salmanian A H, Forghanifard MM, Aghamollaei H. [Design and expression of recombinant HER-2 antigen as a marker for detection of breast cancer]. Pathobiol Res. 2015; 17(4): 88-99. [Article in Persian]
24. Albanell J, Codony J, Rovira A, Mellado B, Gascón P. Mechanism of action of anti-HER2 monoclonal antibodies: scientific update on trastuzumab and 2C4. Adv Exp Med Biol. 2003; 532: 253-68.
25. Sun Y, Feng X, Qu J, Han W, Liu Z, Li X, et al. Expression and Characterization of the Extracellular Domain of Human HER2 from Escherichia Coli, and Production of Polyclonal Antibodies Against the Recombinant Proteins. Appl Biochem Biotechnol. 2015 Jun; 176(4): 1029-43. doi: 10.1007/s12010-015-1627-x
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Kiarostami N, Amani J, Imani Fooladi A A, Mirhosseini A. Effect of recombinant Herceptin on the growth of breast cancer cells. J Gorgan Univ Med Sci 2019; 21 (2) :120-127
URL: http://goums.ac.ir/journal/article-1-3320-en.html
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Volume 21, Issue 2 (7-2019) Back to browse issues page
مجله دانشگاه علوم پزشکی گرگان Journal of Gorgan University of Medical Sciences
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