Matrix metalloproteinase 12 is induced by heterogeneous nuclear ribonucleoprotein K and promotes migration and invasion in nasopharyngeal carcinoma

BMC Cancer

I-Che Chung¹  Lih-Chyang Chen²  An-Ko Chung⁵  Mei Chao⁴  Hsin-Yi Huang⁵  Chuen Hsueh³  Ngan-Ming Tsang⁷  Kai-Ping Chang⁶  Ying Liang¹  Hsin-Pai Li⁴  Yu-Sun Chang⁵ 

[1] Molecular Medicine Research Center, Chang Gung University, 259 Wen-Hwa Ist Road, Taoyuan, Kwei-shan 333, Taiwan

[2] Department of Medicine, Mackay Medical College, No.46, Sec.3, Zhong-Zheng Rd, New Taipei City, San-Zhi Dist 252, Taiwan

[3] Departments of Pathology, Chang Gung Memorial Hospital at Lin-Kou, 5, Fu-Ching St, Taoyuan, Kwei-Shan 333, Taiwan

[4] Department of Microbiology and Immunology, Chang Gung University, 259 Wen-Hwa Ist Road, Taoyuan, Kwei-shan 333, Taiwan

[5] Graduate Institute of Biomedical Sciences, Chang Gung University, 259 Wen-Hwa Ist Road, Taoyuan, Kwei-shan 333, Taiwan

[6] Departments of Otolaryngology, Chang Gung Memorial Hospital at Lin-Kou, 5, Fu-Ching St, Taoyuan, Kwei-Shan 333, Taiwan

[7] Departments of Radiation Oncology, Chang Gung Memorial Hospital at Lin-Kou, 5, Fu-Ching St, Taoyuan, Kwei-Shan 333, Taiwan

关键词: Nasopharyngeal carcinoma;    Invasion;    Migration;    MMP12;    hnRNP K;   

Others  :  858810
DOI  :  10.1186/1471-2407-14-348

 received in 2013-10-16, accepted in 2014-05-06,  发布年份 2014

【 摘 要 】

Background

Overexpression of heterogeneous nuclear ribonucleoprotein K (hnRNP K), a DNA/RNA binding protein, is associated with metastasis in nasopharyngeal carcinoma (NPC). However, the mechanisms underlying hnRNP K-mediated metastasis is unclear. The aim of the present study was to determine the role of matrix metalloproteinase (MMP) in hnRNP K-mediated metastasis in NPC.

Methods

We studied hnRNP K-regulated MMPs by analyzing the expression profiles of MMP family genes in NPC tissues and hnRNP K-knockdown NPC cells using Affymetrix microarray analysis and quantitative RT-PCR. The association of hnRNP K and MMP12 expression in 82 clinically proven NPC cases was determined by immunohistochemical analysis. The hnRNP K-mediated MMP12 regulation was determined by zymography and Western blot, as well as by promoter, DNA pull-down and chromatin immunoprecipitation (ChIP) assays. The functional role of MMP12 in cell migration and invasion was demonstrated by MMP12-knockdown and the treatment of MMP12-specific inhibitor, PF-356231.

Results

MMP12 was overexpressed in NPC tissues, and this high level of expression was significantly correlated with high-level expression of hnRNP K (P = 0.026). The levels of mRNA, protein and enzyme activity of MMP12 were reduced in hnRNP K-knockdown NPC cells. HnRNP K interacting with the region spanning −42 to −33 bp of the transcription start site triggered transcriptional activation of the MMP12 promoter. Furthermore, inhibiting MMP12 by MMP12 knockdown and MMP12-specific inhibitor, PF-356231, significantly reduced the migration and invasion of NPC cells.

Conclusions

Overexpression of MMP12 was significantly correlated with hnRNP K in NPC tissues. HnRNP K can induce MMP12 expression and enzyme activity through activating MMP12 promoter, which promotes cell migration and invasion in NPC cells. In vitro experiments suggest that NPC metastasis with high MMP12 expression may be treated with PF-356231. HnRNP K and MMP12 may be potential therapeutic markers for NPC, but additional validation studies are warranted.

【 授权许可】

   
2014 Chung et al.; licensee BioMed Central Ltd.

【 图 表 】

【 参考文献 】

• [1]Wen F, Shen A, Shanas R, Bhattacharyya A, Lian F, Hostetter G, Shi J: Higher expression of the heterogeneous nuclear ribonucleoprotein k in melanoma. Ann Surg Oncol 2010, 17(10):2619-2627.
• [2]Zhou R, Shanas R, Nelson MA, Bhattacharyya A, Shi J: Increased expression of the heterogeneous nuclear ribonucleoprotein K in pancreatic cancer and its association with the mutant p53. Int J Cancer 2010, 126(2):395-404.
• [3]Barboro P, Repaci E, Rubagotti A, Salvi S, Boccardo S, Spina B, Truini M, Introini C, Puppo P, Ferrari N, Carmignani G, Boccardo F, Balbi C: Heterogeneous nuclear ribonucleoprotein K: altered pattern of expression associated with diagnosis and prognosis of prostate cancer. Br J Cancer 2009, 100(10):1608-1616.
• [4]Matta A, Tripathi SC, DeSouza LV, Grigull J, Kaur J, Chauhan SS, Srivastava A, Thakar A, Shukla NK, Duggal R, DattaGupta S, Ralhan R, MichaelSiu KW: Heterogeneous ribonucleoprotein K is a marker of oral leukoplakia and correlates with poor prognosis of squamous cell carcinoma. Int J Cancer 2009, 125(6):1398-1406.
• [5]Chen LC, Hsueh C, Tsang NM, Liang Y, Chang KP, Hao SP, Yu JS, Chang YS: Heterogeneous ribonucleoprotein k and thymidine phosphorylase are independent prognostic and therapeutic markers for nasopharyngeal carcinoma. Clin Cancer Res 2008, 14(12):3807-3813.
• [6]Choi HS, Hwang CK, Song KY, Law PY, Wei LN, Loh HH: Poly(C)-binding proteins as transcriptional regulators of gene expression. Biochem Biophys Res Commun 2009, 380(3):431-436.
• [7]Chen LC, Chung IC, Hsueh C, Tsang NM, Chi LM, Liang Y, Chen CC, Wang LJ, Chang YS: The antiapoptotic protein, FLIP, is regulated by heterogeneous nuclear ribonucleoprotein K and correlates with poor overall survival of nasopharyngeal carcinoma patients. Cell Death Differ 2010, 17(9):1463-1473.
• [8]Li H, Liu J: Identification of heterogeneous nuclear ribonucleoprotein K as a transactivator for human low density lipoprotein receptor gene transcription. J Biol Chem 2010, 285(23):17789-17797.
• [9]Du Q, Melnikova IN, Gardner PD: Differential effects of heterogeneous nuclear ribonucleoprotein K on Sp1- and Sp3-mediated transcriptional activation of a neuronal nicotinic acetylcholine receptor promoter. J Biol Chem 1998, 273(31):19877-19883.
• [10]Denisenko ON, O'Neill B, Ostrowski J, Van Seuningen I, Bomsztyk K: Zik1, a transcriptional repressor that interacts with the heterogeneous nuclear ribonucleoprotein particle K protein. J Biol Chem 1996, 271(44):27701-27706.
• [11]Chen LC, Liu HP, Li HP, Hsueh C, Yu JS, Liang CL, Chang YS: Thymidine phosphorylase mRNA stability and protein levels are increased through ERK-mediated cytoplasmic accumulation of hnRNP K in nasopharyngeal carcinoma cells. Oncogene 2009, 28(17):1904-1915.
• [12]Notari M, Neviani P, Santhanam R, Blaser BW, Chang JS, Galietta A, Willis AE, Roy DC, Caligiuri MA, Marcucci G, Perrotti D: A MAPK/HNRPK pathway controls BCR/ABL oncogenic potential by regulating MYC mRNA translation. Blood 2006, 107(6):2507-2516.
• [13]Lynch M, Chen L, Ravitz MJ, Mehtani S, Korenblat K, Pazin MJ, Schmidt EV: hnRNP K binds a core polypyrimidine element in the eukaryotic translation initiation factor 4E (eIF4E) promoter, and its regulation of eIF4E contributes to neoplastic transformation. Mol Cell Biol 2005, 25(15):6436-6453.
• [14]Evans JR, Mitchell SA, Spriggs KA, Ostrowski J, Bomsztyk K, Ostarek D, Willis AE: Members of the poly (rC) binding protein family stimulate the activity of the c-myc internal ribosome entry segment in vitro and in vivo. Oncogene 2003, 22(39):8012-8020.
• [15]Murphy G, Nagase H: Progress in matrix metalloproteinase research. Mol Aspects Med 2008, 29(5):290-308.
• [16]Hadler-Olsen E, Winberg JO, Uhlin-Hansen L: Matrix metalloproteinases in cancer: their value as diagnostic and prognostic markers and therapeutic targets. Tumour Biol 2013, 34(4):2041-2051.
• [17]Shuman Moss LA, Jensen-Taubman S, Stetler-Stevenson WG: Matrix metalloproteinases: changing roles in tumor progression and metastasis. Am J Pathol 2012, 181(6):1895-1899.
• [18]Yang R, Xu Y, Li P, Zhang X, Wang J, Gu D, Wang Y: Combined upregulation of matrix metalloproteinase-1 and proteinase-activated receptor-1 predicts unfavorable prognosis in human nasopharyngeal carcinoma. Onco Targets Ther 2013, 6:1139-1146.
• [19]Cui D, Zhang X, Fu Y: Expressions of COX-2 and MMP-2 in nasopharyngeal carcinoma and the their relationship with lymph node metastasis. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi 2008, 22(15):692-694.
• [20]Wong TS, Kwong DL, Sham JS, Wei WI, Kwong YL, Yuen AP: Clinicopathologic significance of plasma matrix metalloproteinase-2 and −9 levels in patients with undifferentiated nasopharyngeal carcinoma. Eur J Surg Oncol 2004, 30(5):560-564.
• [21]Liu Z, Li L, Yang Z, Luo W, Li X, Yang H, Yao K, Wu B, Fang W: Increased expression of MMP9 is correlated with poor prognosis of nasopharyngeal carcinoma. BMC cancer 2010, 10:270. BioMed Central Full Text
• [22]Yoshizaki T, Sato H, Furukawa M, Pagano JS: The expression of matrix metalloproteinase 9 is enhanced by Epstein-Barr virus latent membrane protein 1. Proc Natl Acad Sci U S A 1998, 95(7):3621-3626.
• [23]Lan YY, Hsiao JR, Chang KC, Chang JS, Chen CW, Lai HC, Wu SY, Yeh TH, Chang FH, Lin WH, Su IJ, Chang Y: Epstein-Barr virus latent membrane protein 2A promotes invasion of nasopharyngeal carcinoma cells through ERK/Fra-1-mediated induction of matrix metalloproteinase 9. J Virol 2012, 86(12):6656-6667.
• [24]Yoshizaki T, Sato H, Murono S, Pagano JS, Furukawa M: Matrix metalloproteinase 9 is induced by the Epstein-Barr virus BZLF1 transactivator. Clin Exp Metastasis 1999, 17(5):431-436.
• [25]Kim JM, Kim HJ, Koo BS, Rha KS, Yoon YH: Expression of matrix metalloproteinase-12 is correlated with extracapsular spread of tumor from nodes with metastasis in head and neck squamous cell carcinoma. Eur Arch Otorhinolaryngol 2012, 270(3):1137-1142.
• [26]Ng KT, Qi X, Kong KL, Cheung BY, Lo CM, Poon RT, Fan ST, Man K: Overexpression of matrix metalloproteinase-12 (MMP-12) correlates with poor prognosis of hepatocellular carcinoma. Eur J Cancer 2011, 47(15):2299-2305.
• [27]Hofmann HS, Hansen G, Richter G, Taege C, Simm A, Silber RE, Burdach S: Matrix metalloproteinase-12 expression correlates with local recurrence and metastatic disease in non-small cell lung cancer patients. Clin Cancer Res 2005, 11(3):1086-1092.
• [28]Ding Y, Shimada Y, Gorrin-Rivas MJ, Itami A, Li Z, Hong T, Maeda M, Komoto I, Kawabe A, Kaganoi J, Imamura M: Clinicopathological significance of human macrophage metalloelastase expression in esophageal squamous cell carcinoma. Oncology 2002, 63(4):378-384.
• [29]Yu Y, Chiba Y, Sakai H, Misawa M: Possible involvements of nuclear factor-kappa B and activator protein-1 in the tumor necrosis factor-alpha-induced upregulation of matrix metalloproteinase-12 in human alveolar epithelial A549 cell line. J Pharmacol Sci 2010, 112(1):83-88.
• [30]Wu L, Tanimoto A, Murata Y, Fan J, Sasaguri Y, Watanabe T: Induction of human matrix metalloproteinase-12 gene transcriptional activity by GM-CSF requires the AP-1 binding site in human U937 monocytic cells. Biochem Biophys Res Commun 2001, 285(2):300-307.
• [31]Nabha SM, dos Santos EB, Yamamoto HA, Belizi A, Dong Z, Meng H, Saliganan A, Sabbota A, Bonfil RD, Cher ML: Bone marrow stromal cells enhance prostate cancer cell invasion through type I collagen in an MMP-12 dependent manner. Int J Cancer 2008, 122(11):2482-2490.
• [32]Sarkar S, Nuttall RK, Liu S, Edwards DR, Yong VW: Tenascin-C stimulates glioma cell invasion through matrix metalloproteinase-12. Cancer Res 2006, 66(24):11771-11780.
• [33]Dwivedi A, Slater SC, George SJ: MMP-9 and −12 cause N-cadherin shedding and thereby beta-catenin signalling and vascular smooth muscle cell proliferation. Cardiovasc Res 2009, 81(1):178-186.
• [34]Nocito A, Dahm F, Jochum W, Jang JH, Georgiev P, Bader M, Graf R, Clavien PA: Serotonin regulates macrophage-mediated angiogenesis in a mouse model of colon cancer allografts. Cancer Res 2008, 68(13):5152-5158.
• [35]Goldsmith DB, West TM, Morton R: HLA associations with nasopharyngeal carcinoma in Southern Chinese: a meta-analysis. Clin Otolaryngol Allied Sci 2002, 27(1):61-67.
• [36]Zheng X, Yan L, Nilsson B, Eklund G, Drettner B: Epstein-Barr virus infection, salted fish and nasopharyngeal carcinoma. A case–control study in southern China. Acta oncologica 1994, 33(8):867-872.
• [37]Chen LC, Chen CC, Liang Y, Tsang NM, Chang YS, Hsueh C: A novel role for TNFAIP2: its correlation with invasion and metastasis in nasopharyngeal carcinoma. Mod Pathol 2011, 24(2):175-184.
• [38]Lin CT, Wong CI, Chan WY, Tzung KW, Ho JK, Hsu MM, Chuang SM: Establishment and characterization of two nasopharyngeal carcinoma cell lines. Lab Invest 1990, 62(6):713-724.
• [39]Huang DP, Ho JH, Poon YF, Chew EC, Saw D, Lui M, Li CL, Mak LS, Lai SH, Lau WH: Establishment of a cell line (NPC/HK1) from a differentiated squamous carcinoma of the nasopharynx. Int J Cancer 1980, 26(2):127-132.
• [40]Chen LC, Wang LJ, Tsang NM, Ojcius DM, Chen CC, Ouyang CN, Hsueh C, Liang Y, Chang KP, Chang YS: Tumour inflammasome-derived IL-1beta recruits neutrophils and improves local recurrence-free survival in EBV-induced nasopharyngeal carcinoma. EMBO Mol Med 2012, 4(12):1276-1293.
• [41]Morales R, Perrier S, Florent JM, Beltra J, Dufour S, De Mendez I, Manceau P, Tertre A, Moreau F, Compere D, Dublanchet AC, O'Gara M: Crystal structures of novel non-peptidic, non-zinc chelating inhibitors bound to MMP-12. J Mol Biol 2004, 341(4):1063-1076.
• [42]Terp GE, Cruciani G, Christensen IT, Jorgensen FS: Structural differences of matrix metalloproteinases with potential implications for inhibitor selectivity examined by the GRID/CPCA approach. J Med Chem 2002, 45(13):2675-2684.
• [43]Carpenter B, McKay M, Dundas SR, Lawrie LC, Telfer C, Murray GI: Heterogeneous nuclear ribonucleoprotein K is over expressed, aberrantly localised and is associated with poor prognosis in colorectal cancer. Br J Cancer 2006, 95(7):921-927.
• [44]Wu CS, Chang KP, Chen LC, Chen CC, Liang Y, Hseuh C, Chang YS: Heterogeneous ribonucleoprotein K and thymidine phosphorylase are independent prognostic and therapeutic markers for oral squamous cell carcinoma. Oral oncology 2012, 48(6):516-522.
• [45]Gao R, Yu Y, Inoue A, Widodo N, Kaul SC, Wadhwa R: Heterogeneous nuclear ribonucleoprotein K (hnRNP-K) promotes tumor metastasis by induction of genes involved in extracellular matrix, cell movement, and angiogenesis. J Biol Chem 2013, 288(21):15046-15056.
• [46]Hope NR, Murray GI: The expression profile of RNA-binding proteins in primary and metastatic colorectal cancer: relationship of heterogeneous nuclear ribonucleoproteins with prognosis. Hum Pathol 2011, 42(3):393-402.
• [47]Zhao Y, Jin X, Tian T, Yu DH: Expression of hnRNPK in gastric carcinoma and its relationship with Helicobacter pylori L-form infection. Zhonghua Zhong Liu Za Zhi 2011, 33(10):759-763.
• [48]Lan YY, Yeh TH, Lin WH, Wu SY, Lai HC, Chang FH, Takada K, Chang Y: Epstein-Barr virus Zta upregulates matrix metalloproteinases 3 and 9 that synergistically promote cell invasion in vitro. PloS one 2013, 8(2):e56121.
• [49]Tsai CL, Li HP, Lu YJ, Hsueh C, Liang Y, Chen CL, Tsao SW, Tse KP, Yu JS, Chang YS: Activation of DNA methyltransferase 1 by EBV LMP1 Involves c-Jun NH(2)-terminal kinase signaling. Cancer Res 2006, 66(24):11668-11676.
• [50]Zhang LY, Ho-Fun Lee V, Wong AM, Kwong DL, Zhu YH, Dong SS, Kong KL, Chen J, Tsao SW, Guan XY, Fu L: MicroRNA-144 promotes cell proliferation, migration and invasion in nasopharyngeal carcinoma through repression of PTEN. Carcinogenesis 2013, 34(2):454-463.
• [51]Sato H, Takino T, Miyamori H: Roles of membrane-type matrix metalloproteinase-1 in tumor invasion and metastasis. Cancer Sci 2005, 96(4):212-217.
• [52]Huang MY, Chang HJ, Chung FY, Yang MJ, Yang YH, Wang JY, Lin SR: MMP13 is a potential prognostic marker for colorectal cancer. Oncol Rep 2010, 24(5):1241-1247.
• [53]Kumamoto K, Fujita K, Kurotani R, Saito M, Unoki M, Hagiwara N, Shiga H, Bowman ED, Yanaihara N, Okamura S, Nagashima M, Miyamoto K, Takenoshita S, Yokota J, Harris CC: ING2 is upregulated in colon cancer and increases invasion by enhanced MMP13 expression. Int J Cancer 2009, 125(6):1306-1315.
• [54]Jian P, Yanfang T, Zhuan Z, Jian W, Xueming Z, Jian N: MMP28 (epilysin) as a novel promoter of invasion and metastasis in gastric cancer. BMC cancer 2011, 11:200. BioMed Central Full Text
• [55]Wu HC, Lin YJ, Lee JJ, Liu YJ, Liang ST, Peng Y, Chiu YW, Wu CW, Lin CT: Functional analysis of EBV in nasopharyngeal carcinoma cells. Lab Invest 2003, 83(6):797-812.
• [56]Lo AK, Lo KW, Tsao SW, Wong HL, Hui JW, To KF, Hayward DS, Chui YL, Lau YL, Takada K, Huang DP: Epstein-Barr virus infection alters cellular signal cascades in human nasopharyngeal epithelial cells. Neoplasia 2006, 8(3):173-180.
• [57]Ostareck-Lederer A, Ostareck DH: Precision mechanics with multifunctional tools: how hnRNP K and hnRNPs E1/E2 contribute to post-transcriptional control of gene expression in hematopoiesis. Curr Protein Pept Sci 2012, 13(4):391-400.
• [58]Lyu J, Joo CK: Wnt-7a up-regulates matrix metalloproteinase-12 expression and promotes cell proliferation in corneal epithelial cells during wound healing. J Biol Chem 2005, 280(22):21653-21660.
• [59]Lyu J, Joo CK: Expression of Wnt and MMP in epithelial cells during corneal wound healing. Cornea 2006, 25(10 Suppl 1):S24-S28.
• [60]Samuel S, Twizere JC, Bernstein LR: YB-1 represses AP1-dependent gene transactivation and interacts with an AP-1 DNA sequence. Biochem J 2005, 388(Pt 3):921-928.
• [61]Souissi IJ, Billiet L, Cuaz-Perolin C, Slimane MN, Rouis M: Matrix metalloproteinase-12 gene regulation by a PPAR alpha agonist in human monocyte-derived macrophages. Exp Cell Res 2008, 314(18):3405-3414.
• [62]Moumen A, Masterson P, O'Connor MJ, Jackson SP: hnRNP K: an HDM2 target and transcriptional coactivator of p53 in response to DNA damage. Cell 2005, 123(6):1065-1078.