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Article

A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro

  • Authors:
    • Jikai Yin
    • Zhongliang Cai
    • Li Zhang
    • Jian Zhang
    • Xianli He
    • Xilin Du
    • Qing Wang
    • Jianguo Lu
  • View Affiliations / Copyright

    Affiliations: Department of General Surgery, Tangdu Hospital of The Fourth Military Medical University, Ba'qiao, Xi'an, Shaanxi 710038, P.R. China, Research Institute of General Surgery, Nanjing General Hospital of Nanjing Military Command, Xuan'wu, Nanjing, Jiangsu 210002, P.R. China, Department of Ultrasound Diagnosis, Tangdu Hospital of The Fourth Military Medical University, Ba'qiao, Xi'an, Shaanxi 710038, P.R. China, State Key Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, The Fourth Military Medical University, Ba'qiao, Xi'an, Shaanxi 710032, P.R. China
  • Pages: 1061-1069
    |
    Published online on: January 10, 2013
       https://doi.org/10.3892/ijo.2013.1768
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Abstract

The growth factor receptor bound protein 2 (Grb2) is one of the affirmative targets for cancer therapy, especially for breast cancer. In this study, we hypothesized the Src-homology 2 (SH2) domain in Grb2 may serve as a competitive protein-binding agent to interfere with the proliferation of breast cancer cells in vitro. We designed, constructed, expressed and purified a novel fusion protein containing the protein transduction domain (PTD) and Grb2-SH2 domain (we named it after PTD-Grb2-SH2). An immunofluorescence assay was used to investigate the location of PTD-Grb2-SH2 in cells. MTT assay and EdU experiments were applied to detect the proliferation of breast cancer cells. The ultra-structure was observed using transmission electron microscopy. Flow cytometry was used to determine the cytotoxicity of PTD-Grb2-SH2 on cell proli­feration. We successfully obtained the PTD-Grb2-SH2 fusion protein in soluble form using a prokaryotic expression system. The new fusion protein successfully passed through both the cellular and nuclear membranes of breast cancer cells. The MTT assay showed that PTD-Grb2-SH2 exhibited significant toxicity to breast cancer cells in a dose- and time-dependent manner in vitro. EdU identified the decreased proliferation rates in treated MDA-MB-231 and SK-BR-3 cells. Observation by transmission electron microscopy and flow cytometry further confirmed the cytotoxicity as apoptosis. Our results show that the HIV1-TAT domain is a useful tool for transporting a low molecular weight protein across the cell membrane in vitro. The PTD-Grb2-SH2 may be a novel agent for breast cancer therapy.
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1 

DeSantis C, Siegel R, Bandi P and Jemal A: Breast cancer statistics, 2011. CA Cancer J Clin. 61:409–418. 2011. View Article : Google Scholar

2 

Harris AL, Nicholson S, Sainsbury JR, Farndon J and Wright C: Epidermal growth factor receptors in breast cancer: association with early relapse and death, poor response to hormones and interactions with neu. J Steroid Biochem. 34:123–131. 1989. View Article : Google Scholar : PubMed/NCBI

3 

Sainsbury JR, Malcolm AJ, Appleton DR, Farndon JR and Harris AL: Presence of epidermal growth factor receptor as an indicator of poor prognosis in patients with breast cancer. J Clin Pathol. 38:1225–1228. 1985. View Article : Google Scholar : PubMed/NCBI

4 

Nielsen TO, Hsu FD, Jensen K, et al: Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma. Clin Cancer Res. 10:5367–5374. 2004. View Article : Google Scholar : PubMed/NCBI

5 

Slamon DJ, Godolphin W, Jones LA, et al: Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science. 244:707–712. 1989. View Article : Google Scholar : PubMed/NCBI

6 

Foster CS, Gosden CM and Ke Y: HER2/neu expression in cancer: the pathologist as diagnostician or prophet? Hum Pathol. 34:635–638. 2003. View Article : Google Scholar : PubMed/NCBI

7 

Rosen PP, Lesser ML, Arroyo CD, Cranor M, Borgen P and Norton L: Immunohistochemical detection of HER2/neu in patients with axillary lymph node negative breast carcinoma. A study of epidemiologic risk factors, histologic features, and prognosis. Cancer. 75:1320–1326. 1995. View Article : Google Scholar

8 

Carlomagno C, Perrone F, Gallo C, et al: c-erb B2 overexpression decreases the benefit of adjuvant tamoxifen in early-stage breast cancer without axillary lymph node metastases. J Clin Oncol. 14:2702–2708. 1996.

9 

Gusterson BA, Gelber RD, Goldhirsch A, et al: Prognostic importance of c-erbB-2 expression in breast cancer. International (Ludwig) Breast Cancer Study Group. J Clin Oncol. 10:1049–1056. 1992.PubMed/NCBI

10 

Bianco R, Melisi D, Ciardiello F and Tortora G: Key cancer cell signal transduction pathways as therapeutic targets. Eur J Cancer. 42:290–294. 2006. View Article : Google Scholar : PubMed/NCBI

11 

Chardin P, Cussac D, Maignan S and Ducruix A: The Grb2 adaptor. FEBS Lett. 369:47–51. 1995. View Article : Google Scholar : PubMed/NCBI

12 

Seger R and Krebs EG: The MAPK signaling cascade. FASEB J. 9:726–735. 1995.PubMed/NCBI

13 

Di Fulvio M, Henkels KM and Gomez-Cambronero J: Short-hairpin RNA-mediated stable silencing of Grb2 impairs cell growth and DNA synthesis. Biochem Biophys Res Commun. 357:737–742. 2007.PubMed/NCBI

14 

Machida K and Mayer BJ: The SH2 domain: versatile signaling module and pharmaceutical target. Biochim Biophys Acta. 1747:1–25. 2005. View Article : Google Scholar : PubMed/NCBI

15 

Pawson T: Protein modules and signalling networks. Nature. 373:573–580. 1995. View Article : Google Scholar : PubMed/NCBI

16 

Mayer BJ, Jackson PK and Baltimore D: The noncatalytic src homology region 2 segment of abl tyrosine kinase binds to tyrosine-phosphorylated cellular proteins with high affinity. Proc Natl Acad Sci USA. 88:627–631. 1991. View Article : Google Scholar : PubMed/NCBI

17 

Mayer BJ, Jackson PK, Van Etten RA and Baltimore D: Point mutations in the abl SH2 domain coordinately impair phosphotyrosine binding in vitro and transforming activity in vivo. Mol Cell Biol. 12:609–618. 1992.PubMed/NCBI

18 

Schwarze SR, Hruska KA and Dowdy SF: Protein transduction: unrestricted delivery into all cells? Trends Cell Biol. 10:290–295. 2000. View Article : Google Scholar : PubMed/NCBI

19 

Lindgren M, Hallbrink M, Prochiantz A and Langel U: Cell-penetrating peptides. Trends Pharmacol Sci. 21:99–103. 2000. View Article : Google Scholar

20 

Liang Y, Sun Q, Jiang S, et al: Construction and expression of a vector containing protein transduction domain and bcr/abl fusion gene. Zhonghua Xue Ye Xue Za Zhi. 23:5–8. 2002.(In Chinese).

21 

Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227:680–685. 1970. View Article : Google Scholar : PubMed/NCBI

22 

Sangadala S, Okada M, Liu Y, Viggeswarapu M, Titus L and Boden SD: Engineering, cloning, and functional characterization of recombinant LIM mineralization protein-1 containing an N-terminal HIV-derived membrane transduction domain. Protein Expr Purif. 65:165–173. 2009. View Article : Google Scholar

23 

Yarden Y and Sliwkowski MX: Untangling the ErbB signalling network. Nat Rev Mol Cell Biol. 2:127–137. 2001. View Article : Google Scholar : PubMed/NCBI

24 

Lung FD and Tsai JY: Grb2 SH2 domain-binding peptide analogs as potential anticancer agents. Biopolymers. 71:132–140. 2003. View Article : Google Scholar : PubMed/NCBI

25 

Kim D, Jeon C, Kim JH, et al: Cytoplasmic transduction peptide (CTP): new approach for the delivery of biomolecules into cytoplasm in vitro and in vivo. Exp Cell Res. 312:1277–1288. 2006. View Article : Google Scholar : PubMed/NCBI

26 

Schwarze SR, Ho A, Vocero-Akbani A and Dowdy SF: In vivo protein transduction: delivery of a biologically active protein into the mouse. Science. 285:1569–1572. 1999. View Article : Google Scholar : PubMed/NCBI

27 

Tan M, Lan KH, Yao J, et al: Selective inhibition of ErbB2-overexpressing breast cancer in vivo by a novel TAT-based ErbB2-targeting signal transducers and activators of transcription 3-blocking peptide. Cancer Res. 66:3764–3772. 2006. View Article : Google Scholar : PubMed/NCBI

28 

Harada H, Kizaka-Kondoh S and Hiraoka M: Antitumor protein therapy; application of the protein transduction domain to the development of a protein drug for cancer treatment. Breast Cancer. 13:16–26. 2006. View Article : Google Scholar : PubMed/NCBI

29 

Sorriento D, Campanile A, Santulli G, et al: A new synthetic protein, TAT-RH, inhibits tumor growth through the regulation of NFkappaB activity. Mol Cancer. 8:972009. View Article : Google Scholar : PubMed/NCBI

30 

Liang YJS, Han H, Liu L, Sun Q, Chen R, Wu R, Du J and Li Q: TAT PTD-BCR/ABL SH3 fusion protein induces the apoptosis of K562 leukemic cell line. J Fourth Mil Med Univ. 23:42002.

31 

Yin JK, Liang YM, He XL, et al: Fusion protein containing SH3 domain of c-Abl induces hepatocarcinoma cells to apoptosis. Hepatol Res. 37:454–463. 2007. View Article : Google Scholar : PubMed/NCBI

32 

Ogura K, Shiga T, Yokochi M, Yuzawa S, Burke TR Jr and Inagaki F: Solution structure of the Grb2 SH2 domain complexed with a high-affinity inhibitor. J Biomol NMR. 42:197–207. 2008. View Article : Google Scholar : PubMed/NCBI

33 

Dierck K, Machida K, Mayer BJ and Nollau P: Profiling the tyrosine phosphorylation state using SH2 domains. Methods Mol Biol. 527:131–155. 2009. View Article : Google Scholar : PubMed/NCBI

34 

Jiang X, Huang F, Marusyk A and Sorkin A: Grb2 regulates internalization of EGF receptors through clathrin-coated pits. Mol Biol Cell. 14:858–870. 2003. View Article : Google Scholar : PubMed/NCBI

35 

Yamazaki T, Zaal K, Hailey D, Presley J, Lippincott-Schwartz J and Samelson LE: Role of Grb2 in EGF-stimulated EGFR internalization. J Cell Sci. 115:1791–1802. 2002.PubMed/NCBI

36 

Burke P, Schooler K and Wiley HS: Regulation of epidermal growth factor receptor signaling by endocytosis and intracellular trafficking. Mol Biol Cell. 12:1897–1910. 2001. View Article : Google Scholar : PubMed/NCBI

37 

Schulze WX, Deng L and Mann M: Phosphotyrosine inter-actome of the ErbB-receptor kinase family. Mol Syst Biol. 1:2005.0008. 2005. View Article : Google Scholar : PubMed/NCBI

38 

Seiden-Long I, Navab R, Shih W, et al: Gab1 but not Grb2 mediates tumor progression in Met overexpressing colorectal cancer cells. Carcinogenesis. 29:647–655. 2008.PubMed/NCBI

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Copy and paste a formatted citation
Spandidos Publications style
Yin J, Cai Z, Zhang L, Zhang J, He X, Du X, Wang Q and Lu J: A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro. Int J Oncol 42: 1061-1069, 2013.
APA
Yin, J., Cai, Z., Zhang, L., Zhang, J., He, X., Du, X. ... Lu, J. (2013). A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro. International Journal of Oncology, 42, 1061-1069. https://doi.org/10.3892/ijo.2013.1768
MLA
Yin, J., Cai, Z., Zhang, L., Zhang, J., He, X., Du, X., Wang, Q., Lu, J."A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro". International Journal of Oncology 42.3 (2013): 1061-1069.
Chicago
Yin, J., Cai, Z., Zhang, L., Zhang, J., He, X., Du, X., Wang, Q., Lu, J."A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro". International Journal of Oncology 42, no. 3 (2013): 1061-1069. https://doi.org/10.3892/ijo.2013.1768
Copy and paste a formatted citation
x
Spandidos Publications style
Yin J, Cai Z, Zhang L, Zhang J, He X, Du X, Wang Q and Lu J: A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro. Int J Oncol 42: 1061-1069, 2013.
APA
Yin, J., Cai, Z., Zhang, L., Zhang, J., He, X., Du, X. ... Lu, J. (2013). A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro. International Journal of Oncology, 42, 1061-1069. https://doi.org/10.3892/ijo.2013.1768
MLA
Yin, J., Cai, Z., Zhang, L., Zhang, J., He, X., Du, X., Wang, Q., Lu, J."A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro". International Journal of Oncology 42.3 (2013): 1061-1069.
Chicago
Yin, J., Cai, Z., Zhang, L., Zhang, J., He, X., Du, X., Wang, Q., Lu, J."A recombined fusion protein PTD-Grb2-SH2 inhibits the proliferation of breast cancer cells in vitro". International Journal of Oncology 42, no. 3 (2013): 1061-1069. https://doi.org/10.3892/ijo.2013.1768
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