Increased phosphorylation of histone H3 at serine 10 is involved in Epstein-Barr virus latent membrane protein-1-induced carcinogenesis of nasopharyngeal carcinoma

BMC Cancer

Binbin Li³  Guoliang Huang³  Xiangning Zhang²  Rong Li²  Jian Wang³  Ziming Dong¹  Zhiwei He³ 

[1] Department of Pathophysiology, Basic Medical College of Zhengzhou University, No.100 of Science Road, Zhengzhou, 450001, China

[2] Department of Pathophysiology, Guangdong Medical College, Guangdong, Dongguan, 523808, China

[3] Key Laboratory for Medical Diagnostics of Guangdong Province, Sino-American Cancer Research Institute, Guangdong Medical College, No. 1 Xincheng Road, Guangdong, Dongguan, 523808, China

关键词: MSK1;    Epstein-Barr virus latent membrane protein-1;    Nasopharyngeal carcinoma;    Histone H3 phosphorylation;   

Others  :  1079866
DOI  :  10.1186/1471-2407-13-124

 received in 2013-01-07, accepted in 2013-03-11,  发布年份 2013

【 摘 要 】

Background

Increased histone H3 phosphorylation is an essential regulatory mechanism for neoplastic cell transformation. We aimed to explore the role of histone H3 phosphorylation at serine10 (p-H3Ser10) in Epstein-Barr virus (EBV) latent membrane protein-1 (LMP1)-induced carcinogenesis of nasopharyngeal carcinoma (NPC).

Methods

The expression of p-H3Ser10 was detected by the immunohistochemical analysis in NPC, chronic nasopharyngitis and normal nasopharynx tissues, and its correlation with LMP1 was analyzed in NPC tissues and cell lines. Using the small interfering RNA (siRNA)-H3 and histone H3 mutant (S10A), the effect of histone H3 Ser10 motif on LMP1-induced CNE1 cell proliferation, transformation and activator protein-1 (AP-1) activation were evaluated by CCK-8, focus-forming and reporter gene assay respectively. Mitogen- and stress-activated kinase 1 (MSK1) kinase activity and phosphorylation were detected by in vitro kinase assay and western blot. Using MSK1 inhibitor H89 or siRNA-MSK1, the regulatory role of MSK1 on histone H3 phosphorylation and AP-1 activation were analyzed.

Results

Immunohistochemical analysis revealed that the expression of p-H3Ser10 was significantly higher in the poorly differentiated NPC tissues than that in chronic nasopharyngitis (p <0.05) and normal nasopharynx tissues (p <0.001). Moreover, high level of p-H3Ser10 was positively correlated with the expression of LMP1 in NPC tissues (χ²=6.700, p =0.01; C=0.350) and cell lines. The knockdown and mutant (S10A) of histone H3 suppressed LMP1-induced CNE1 cell proliferation, foci formation and AP-1 activation. In addition, LMP1 could increase MSK1 kinase activity and phosphorylation. MSK1 inhibitor H89 or knockdown of MSK1 by siRNA blocked LMP1-induced phosphorylation of histone H3 at Ser10 and AP-1 activation.

Conclusion

EBV-LMP1 can induce phosphorylation of histone H3 at Ser10 via MSK1. Increased phosphorylation of histone H3 at Ser10 is likely a crucial regulatory mechanism involved in LMP1-induced carcinogenesis of NPC.

【 授权许可】

   
2013 Li et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Kouzarides T: Chromatin modifications and their function. Cell 2007, 128(4):693-705.
• [2]Gurley LR, Walters RA, Tobey RA: Cell cycle-specific changes in histone phosphorylation associated with cell proliferation and chromosome condensation. J Cell Biol 1974, 60(2):356-364.
• [3]Mahadevan LC, Willis AC, Barratt MJ: Rapid histone H3 phosphorylation in response to growth factors, phorbol esters, okadaic acid, and protein synthesis inhibitors. Cell 1991, 65(5):775-783.
• [4]Hazzalin CA, Cano E, Cuenda A, Barratt MJ, Cohen P, Mahadevan LC: p38/RK is essential for stress-induced nuclear responses: JNK/SAPKs and c-Jun/ATF-2 phosphorylation are insufficient. Curr Biol 1996, 6(8):1028-1031.
• [5]McMahon SB, Monroe JG: Role of primary response genes in generating cellular responses to growth factors. FASEB J 1992, 6(9):2707-2715.
• [6]Chadee DN, Hendzel MJ, Tylipski CP, Allis CD, Bazett-Jones DP, Wright JA, Davie JR: Increased Ser-10 phosphorylation of histone H3 in mitogen-stimulated and oncogene-transformed mouse fibroblasts. J Biol Chem 1999, 274(35):24914-24920.
• [7]Strelkov IS, Davie JR: Ser-10 phosphorylation of histone H3 and immediate early gene expression in oncogene-transformed mouse fibroblasts. Cancer Res 2002, 62(1):75-78.
• [8]Sassone-Corsi P, Mizzen CA, Cheung P, Crosio C, Monaco L, Jacquot S, Hanauer A, Allis CD: Requirement of Rsk-2 for epidermal growth factor-activated phosphorylation of histone H3. Science 1999, 285(5429):886-891.
• [9]Drobic B, Espino PS, Davie JR: Mitogen- and stress-activated protein kinase 1 activity and histone h3 phosphorylation in oncogene-transformed mouse fibroblasts. Cancer Res 2004, 64(24):9076-9079.
• [10]Kim HG, Lee KW, Cho YY, Kang NJ, Oh SM, Bode AM, Dong Z: Mitogen- and stress-activated kinase 1-mediated histone H3 phosphorylation is crucial for cell transformation. Cancer Res 2008, 68(7):2538-2547.
• [11]Tange S, Ito S, Senga T, Hamaguchi M: Phosphorylation of histone H3 at Ser10: its role in cell transformation by v-Src. Biochem Biophys Res Commun 2009, 386(4):588-592.
• [12]He Z, Ma WY, Liu G, Zhang Y, Bode AM, Dong Z: Arsenite-induced phosphorylation of histone H3 at serine 10 is mediated by Akt1, extracellular signal-regulated kinase 2, and p90 ribosomal S6 kinase 2 but not mitogen- and stress-activated protein kinase 1. J Biol Chem 2003, 278(12):10588-10593.
• [13]Raab-Traub N: Epstein-Barr virus in the pathogenesis of NPC. Semin Cancer Biol 2002, 12(6):431-441.
• [14]Kaye KM, Izumi KM, Mosialos G, Kieff E: The Epstein-Barr virus LMP1 cytoplasmic carboxy terminus is essential for B-lymphocyte transformation; fibroblast cocultivation complements a critical function within the terminal 155 residues. J Virol 1995, 69(2):675-683.
• [15]Moorthy RK, Thorley-Lawson DA: All three domains of the Epstein-Barr virus-encoded latent membrane protein LMP-1 are required for transformation of rat-1 fibroblasts. J Virol 1993, 67(3):1638-1646.
• [16]Hu LF, Chen F, Zheng X, Ernberg I, Cao SL, Christensson B, Klein G, Winberg G: Clonability and tumorigenicity of human epithelial cells expressing the EBV encoded membrane protein LMP1. Oncogene 1993, 8(6):1575-1583.
• [17]Zheng H, Li LL, Hu DS, Deng XY, Cao Y: Role of Epstein-Barr virus encoded latent membrane protein 1 in the carcinogenesis of nasopharyngeal carcinoma. Cell Mol Immunol 2007, 4(3):185-196.
• [18]Morris MA, Dawson CW, Young LS: Role of the Epstein-Barr virus-encoded latent membrane protein-1, LMP1, in the pathogenesis of nasopharyngeal carcinoma. Future Oncol 2009, 5(6):811-825.
• [19]Eliopoulos AG, Young LS: LMP1 structure and signal transduction. Semin Cancer Biol 2001, 11(6):435-444.
• [20]Chen Y: Chen XY: [Effect of Epstein-Barr virus latent membrane protein 1 (LMP1) on apoptosis of nasopharyngeal carcinoma cell line CNE1]. Ai Zheng 2002, 21(5):498-503.
• [21]Kieser A, Kilger E, Gires O, Ueffing M, Kolch W, Hammerschmidt W: Epstein-Barr virus latent membrane protein-1 triggers AP-1 activity via the c-Jun N-terminal kinase cascade. EMBO J 1997, 16(21):6478-6485.
• [22]Horikawa T, Sheen TS, Takeshita H, Sato H, Furukawa M, Yoshizaki T: Induction of c-Met proto-oncogene by Epstein-Barr virus latent membrane protein-1 and the correlation with cervical lymph node metastasis of nasopharyngeal carcinoma. Am J Pathol 2001, 159(1):27-33.
• [23]Sistayanarain A, Tsuneyama K, Zheng H, Takahashi H, Nomoto K, Cheng C, Murai Y, Tanaka A, Takano Y: Expression of Aurora-B kinase and phosphorylated histone H3 in hepatocellular carcinoma. Anticancer Res 2006, 26(5A):3585-3593.
• [24]Shechter D, Dormann HL, Allis CD, Hake SB: Extraction, purification and analysis of histones. Nat Protoc 2007, 2(6):1445-1457.
• [25]Angel P, Karin M: The role of Jun, Fos and the AP-1 complex in cell-proliferation and transformation. Biochim Biophys Acta 1991, 1072(2–3):129-157.
• [26]Liu Y, Ludes-Meyers J, Zhang Y, Munoz-Medellin D, Kim HT, Lu C, Ge G, Schiff R, Hilsenbeck SG, Osborne CK: Inhibition of AP-1 transcription factor causes blockade of multiple signal transduction pathways and inhibits breast cancer growth. Oncogene 2002, 21(50):7680-7689.
• [27]Ota T, Suto S, Katayama H, Han ZB, Suzuki F, Maeda M, Tanino M, Terada Y, Tatsuka M: Increased mitotic phosphorylation of histone H3 attributable to AIM-1/Aurora-B overexpression contributes to chromosome number instability. Cancer Res 2002, 62(18):5168-5177.
• [28]Sundar IK, Chung S, Hwang JW, Lapek JD Jr, Bulger M, Friedman AE, Yao H, Davie JR, Rahman I: Mitogen- and stress-activated kinase 1 (MSK1) regulates cigarette smoke-induced histone modifications on NF-kappaB-dependent genes. PLoS One 2012, 7(2):e31378.
• [29]Ke Q, Li Q, Ellen TP, Sun H, Costa M: Nickel compounds induce phosphorylation of histone H3 at serine 10 by activating JNK-MAPK pathway. Carcinogenesis 2008, 29(6):1276-1281.
• [30]Lee S, Horn V, Julien E, Liu Y, Wysocka J, Bowerman B, Hengartner MO, Herr W: Epigenetic regulation of histone H3 serine 10 phosphorylation status by HCF-1 proteins in C. elegans and mammalian cells. PLoS One 2007, 2(11):1213.
• [31]Gires O, Zimber-Strobl U, Gonnella R, Ueffing M, Marschall G, Zeidler R, Pich D, Hammerschmidt W: Latent membrane protein 1 of Epstein-Barr virus mimics a constitutively active receptor molecule. EMBO J 1997, 16(20):6131-6140.
• [32]Mosialos G, Birkenbach M, Yalamanchili R, VanArsdale T, Ware C, Kieff E: The Epstein-Barr virus transforming protein LMP1 engages signaling proteins for the tumor necrosis factor receptor family. Cell 1995, 80(3):389-399.
• [33]Choi HS, Choi BY, Cho YY, Mizuno H, Kang BS, Bode AM, Dong Z: Phosphorylation of histone H3 at serine 10 is indispensable for neoplastic cell transformation. Cancer Res 2005, 65(13):5818-5827.
• [34]Vermeulen L, De Wilde G, Van Damme P, Vanden Berghe W, Haegeman G: Transcriptional activation of the NF-kappaB p65 subunit by mitogen- and stress-activated protein kinase-1 (MSK1). EMBO J 2003, 22(6):1313-1324.
• [35]Wiggin GR, Soloaga A, Foster JM, Murray-Tait V, Cohen P, Arthur JS: MSK1 and MSK2 are required for the mitogen- and stress-induced phosphorylation of CREB and ATF1 in fibroblasts. Mol Cell Biol 2002, 22(8):2871-2881.
• [36]Zhang Y, Liu G, Dong Z: MSK1 and JNKs mediate phosphorylation of STAT3 in UVA-irradiated mouse epidermal JB6 cells. J Biol Chem 2001, 276(45):42534-42542.
• [37]Calipel A, Lefevre G, Pouponnot C, Mouriaux F, Eychene A, Mascarelli F: Mutation of B-Raf in human choroidal melanoma cells mediates cell proliferation and transformation through the MEK/ERK pathway. J Biol Chem 2003, 278(43):42409-42418.
• [38]Dunn KL, Espino PS, Drobic B, He S, Davie JR: The Ras-MAPK signal transduction pathway, cancer and chromatin remodeling. Biochem Cell Biol 2005, 83(1):1-14.
• [39]Soloaga A, Thomson S, Wiggin GR, Rampersaud N, Dyson MH, Hazzalin CA, Mahadevan LC, Arthur JS: MSK2 and MSK1 mediate the mitogen- and stress-induced phosphorylation of histone H3 and HMG-14. EMBO J 2003, 22(11):2788-2797.
• [40]Song X, Tao YG, Deng XY, Jin X, Tan YN, Tang M, Wu Q, Lee LM, Cao Y: Heterodimer formation between c-Jun and Jun B proteins mediated by Epstein-Barr virus encoded latent membrane protein 1. Cell Signal 2004, 16(10):1153-1162.
• [41]Song X, Tao Y, Zeng L, Yang J, Tang F, Lee LM, Gong J, Wu Q, Cao Y: Epstein-Barr virus-encoded latent membrane protein 1 modulates cyclin D1 by c-Jun/Jun B heterodimers. Sci China C Life Sci 2005, 48(4):385-393.
• [42]Chiu R, Boyle WJ, Meek J, Smeal T, Hunter T, Karin M: The c-Fos protein interacts with c-Jun/AP-1 to stimulate transcription of AP-1 responsive genes. Cell 1988, 54(4):541-552.