Activation of STAT5 contributes to proliferation in U87 human glioblastoma multiforme cells

Feng,Cui; Cao,Shouqiang

doi:10.3892/mmr.2014.2223

Activation of STAT5 contributes to proliferation in U87 human glioblastoma multiforme cells

Authors:
- Cui Feng
- Shouqiang Cao
View Affiliations

Affiliations: Department of Anesthesiology, Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China, Department of Neurosurgery, Fourth Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China
Published online on: May 8, 2014 https://doi.org/10.3892/mmr.2014.2223
Pages: 203-210

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Rapid increases in the tyrosine phosphorylation of signal transducers and activators of transcription 5 (STAT5) proteins have been extensively documented in cells stimulated with cytokines and growth factors. However, the mechanisms by which STAT5 translocates to the nucleus and regulates proliferation in human glioblastoma multiforme cells have not been studied in detail. To the best of our knowledge, the present study demonstrated for first time that stimulation of a glioblastoma multiforme (GBM) cell line (U87-MG) with hepatocyte growth factor (HGF) resulted in the phosphorylation of STAT5 at Tyr-694/699 and nuclear translocation of STAT5. In addition, HGF promoted nuclear translocation of STAT5 in a time-dependent manner and increased the proliferation of U87-MG cells. In order to determine the role of STAT5 directly, RNA interference was used to knockdown STAT5 expression in the U87-MG cell line. It was illustrated that small interfering RNA (siRNA) against STAT5 successfully inhibited the protein expression of STAT5 in the U87-MG cell line, leading to a potent suppression of tumor cell proliferation with or without HGF treatment. In order to broaden the investigation and to determine the role of STAT5 in vivo, immunohistochemistry (IHC) was applied to evaluate STAT5 expression in 100 newly diagnosed glioma and 10 non-neoplastic brain tissues. p-STAT5 expression increased according to the histopathological grade of the glioma. However, no p-STAT5 staining was observed in non-neoplastic brain tissues. These findings suggested that inhibition of the STAT5 pathway may be an effective therapeutic strategy for the clinical management of GBM.

View Figures

View References

1	Buitenhuis M, Coffer PJ and Koenderman L: Signal transducer and activator of transcription 5 (STAT5). Int J Biochem Cell Biol. 36:2120–2124. 2004. View Article : Google Scholar : PubMed/NCBI
2	Cao S, Wang C, Zheng Q, et al: STAT5 regulates glioma cell invasion by pathways dependent and independent of STAT5 DNA binding. Neurosci Lett. 487:228–233. 2011. View Article : Google Scholar : PubMed/NCBI
3	Zhang H, Li M, Han Y, et al: Down-regulation of miR-27a might reverse multidrug resistance of esophageal squamous cell carcinoma. Dig Dis Sci. 55:2545–2551. 2010. View Article : Google Scholar : PubMed/NCBI
4	Wang C, Cao S, Yan Y, et al: TLR9 expression in glioma tissues correlated to glioma progression and the prognosis of GBM patients. BMC Cancer. 10:4152010. View Article : Google Scholar : PubMed/NCBI
5	Heim MH: The Jak-STAT pathway: cytokine signalling from the receptor to the nucleus. J Recept Signal Transduct Res. 19:75–120. 1999. View Article : Google Scholar : PubMed/NCBI
6	Hou J, Schindler U, Henzel WJ, Wong SC and McKnight SL: Identification and purification of human Stat proteins activated in response to interleukin-2. Immunity. 2:321–329. 1995. View Article : Google Scholar : PubMed/NCBI
7	Mui AL, Wakao H, O’Farrell AM, Harada N and Miyajima A: Interleukin-3, granulocyte-macrophage colony stimulating factor and interleukin-5 transduce signals through two STAT5 homologs. EMBO J. 14:1166–1175. 1995.
8	Foxwell BM, Beadling C, Guschin D, Kerr I and Cantrell D: Interleukin-7 can induce the activation of Jak 1, Jak 3 and STAT 5 proteins in murine T cells. Eur J Immunol. 25:3041–3046. 1995. View Article : Google Scholar : PubMed/NCBI
9	Barahmand-pour F, Meinke A, Eilers A, Gouilleux F, Groner B and Decker T: Colony-stimulating factors and interferon-gamma activate a protein related to MGF-Stat 5 to cause formation of the differentiation-induced factor in myeloid cells. FEBS Lett. 360:29–33. 1995. View Article : Google Scholar
10	Wartmann M, Cella N, Hofer P, et al: Lactogenic hormone activation of Stat5 and transcription of the beta-casein gene in mammary epithelial cells is independent of p42 ERK2 mitogen-activated protein kinase activity. J Biol Chem. 271:31863–31868. 1996. View Article : Google Scholar
11	Pallard C, Gouilleux F, Bénit L, et al: Thrombopoietin activates a STAT5-like factor in hematopoietic cells. EMBO J. 14:2847–2856. 1995.PubMed/NCBI
12	Joung YH, Lee MY, Lim EJ, et al: Hypoxia activates the IGF-1 expression through STAT5b in human HepG2 cells. Biochem Biophys Res Commun. 358:733–738. 2007. View Article : Google Scholar : PubMed/NCBI
13	Sultan AS, Xie J, LeBaron MJ, Ealley EL, Nevalainen MT and Rui H: Stat5 promotes homotypic adhesion and inhibits invasive characteristics of human breast cancer cells. Oncogene. 24:746–760. 2005. View Article : Google Scholar : PubMed/NCBI
14	Weaver AM and Silva CM: Modulation of signal transducer and activator of transcription 5b activity in breast cancer cells by mutation of tyrosines within the transactivation domain. Mol Endocrinol. 20:2392–2405. 2006. View Article : Google Scholar
15	Xi S, Zhang Q, Dyer KF, et al: Src kinases mediate STAT growth pathways in squamous cell carcinoma of the head and neck. J Biol Chem. 278:31574–31583. 2003. View Article : Google Scholar : PubMed/NCBI
16	Xi S, Zhang Q, Gooding WE, Smithgall TE and Grandis JR: Constitutive activation of Stat5b contributes to carcinogenesis in vivo. Cancer Res. 63:6763–6771. 2003.PubMed/NCBI
17	Cao S, Yan Y, Zhang X, et al: EGF stimulates cyclooxygenase-2 expression through the STAT5 signaling pathway in human lung adenocarcinoma A549 cells. Int J Oncol. 39:383–391. 2011.PubMed/NCBI
18	Spiekermann K, Biethahn S, Wilde S, Hiddemann W and Alves F: Constitutive activation of STAT transcription factors in acute myelogenous leukemia. Eur J Haematol. 67:63–71. 2001. View Article : Google Scholar : PubMed/NCBI
19	Chin H, Nakamura N, Kamiyama R, Miyasaka N, Ihle JN and Miura O: Physical and functional interactions between Stat5 and the tyrosine-phosphorylated receptors for erythropoietin and interleukin-3. Blood. 88:4415–4425. 1996.PubMed/NCBI
20	Pircher TJ, Petersen H, Gustafsson JA and Haldosén LA: Extracellular signal-regulated kinase (ERK) interacts with signal transducer and activator of transcription (STAT) 5a. Mol Endocrinol. 13:555–565. 1999. View Article : Google Scholar : PubMed/NCBI
21	Koppikar P, Lui VW, Man D, et al: Constitutive activation of signal transducer and activator of transcription 5 contributes to tumor growth, epithelial-mesenchymal transition, and resistance to epidermal growth factor receptor targeting. Clin Cancer Res. 14:7682–7690. 2008. View Article : Google Scholar
22	Xiong H, Su WY, Liang QC, et al: Inhibition of STAT5 induces G1 cell cycle arrest and reduces tumor cell invasion in human colorectal cancer cells. Lab Invest. 89:717–725. 2009. View Article : Google Scholar : PubMed/NCBI