The Epstein-Barr Virus-Encoded MicroRNA MiR-BART9 Promotes Tumor Metastasis by Targeting E-Cadherin in Nasopharyngeal Carcinoma

English version České info

MicroRNAs (miRNAs) are a family of small RNA molecules that negatively regulate the expression of protein-coding genes and play critical roles in orchestrating diverse cellular processes. This regulatory mechanism is also exploited by viruses to direct their life cycle and evade the host immune system. Epstein-Barr virus (EBV) is an oncogenic virus that is closely associated with multiple human diseases, including nasopharyngeal carcinoma (NPC), which is a highly metastatic type of tumor and is frequently reported in South Asia. Several viral proteins have been found to promote the migration and invasiveness of NPC cells. However, not all tumor tissues express these viral oncoproteins, suggesting that other mechanisms may contribute to the aggressive behavior of NPC tumor cells. A previous sequencing study by our group revealed that the EBV miRNA miR-BART9 was expressed at high levels in all EBV-positive NPC tissues. In the present study, we used gain -⁠ and loss-of-function approaches to investigate the effect of miR-BART9 in EBV-negative and EBV-positive NPC cells. We discovered that miR-BART9 promotes the migration and invasiveness of cultured NPC cells. The promigratory activity observed in vitro was manifested as an enhanced metastatic ability in vivo. Computational analysis revealed that miR-BART9 may target E-cadherin, a membrane protein that is pivotal in preserving cell-cell junctions and the epithelial phenotype. Through biochemical assays and functional rescue analysis, we confirmed that miR-BART9 specifically inhibits E-cadherin to induce a mesenchymal-like phenotype and promote the migration of NPC cells. These results indicated that miR-BART9 is a prometastatic viral miRNA and suggested that high levels of miR-BART9 in EBV-positive NPC cells may contribute to the aggressiveness of tumor cells.

Vyšlo v časopise: The Epstein-Barr Virus-Encoded MicroRNA MiR-BART9 Promotes Tumor Metastasis by Targeting E-Cadherin in Nasopharyngeal Carcinoma. PLoS Pathog 10(2): e32767. doi:10.1371/journal.ppat.1003974
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003974

Souhrn

Zdroje

1. KleinE, KisLL, KleinG (2007) Epstein-Barr virus infection in humans: from harmless to life endangering virus-lymphocyte interactions. Oncogene 26 (9) 1297–305.

2. TaoQ, YoungLS, WoodmanCB, MurrayPG (2006) Epstein-Barr virus (EBV) and its associated human cancers–genetics, epigenetics, pathobiology and novel therapeutics. Front Biosci 11 : 2672–713.

3. Bar-SelaG, KutenA, MinkovI, Gov-AriE, Ben-IzhakO (2004) Prevalence and relevance of EBV latency in nasopharyngeal carcinoma in Israel. J Clin Pathol 57 (3) 290–3.

4. MirzamaniN, SalehianP, FarhadiM, TehranEA (2006) Detection of EBV and HPV in nasopharyngeal carcinoma by in situ hybridization. Exp Mol Pathol 81 (3) 231–4.

5. TsaoSW, TramoutanisG, DawsonCW, LoAK, HuangDP (2002) The significance of LMP1 expression in nasopharyngeal carcinoma. Semin Cancer Biol 12 (6) 473–87.

6. WeiMX, OokaT (1989) A transforming function of the BARF1 gene encoded by Epstein-Barr virus. EMBO J 8 (10) 2897–903.

7. AllenMD, YoungLS, DawsonCW (2005) The Epstein-Barr virus-encoded LMP2A and LMP2B proteins promote epithelial cell spreading and motility. J Virol 79 (3) 1789–802.

8. AltmannM, PichD, RuissR, WangJ, SugdenB, et al. (2006) Transcriptional activation by EBV nuclear antigen 1 is essential for the expression of EBV's transforming genes. Proc Natl Acad Sci U S A 103 (38) 14188–93.

9. PfefferS, ZavolanM, GrässerFA, ChienM, RussoJJ, et al. (2004) Identification of virus-encoded microRNAs. Science 304 (5671) 734–6.

10. AmbrosV (2004) The functions of animal microRNAs. Nature 431 (7006) 350–5.

11. BartelDP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116 (2) 281–97.

12. ChenCZ (2005) MicroRNAs as oncogenes and tumor suppressors. N Engl J Med 353 (17) 1768–71.

13. ZhuJY, PfuhlT, MotschN, BarthS, NichollsJ, et al. (2009) Identification of novel Epstein-Barr virus microRNA genes from nasopharyngeal carcinomas. J Virol 83 (7) 3333–41.

14. PrattZL, KuzembayevaM, SenguptaS, SugdenB (2009) The microRNAs of Epstein-Barr Virus are expressed at dramatically differing levels among cell lines. Virology 386 (2) 387–97.

15. CaiX, SchäferA, LuS, BilelloJP, DesrosiersRC, et al. (2006) Epstein-Barr virus microRNAs are evolutionarily conserved and differentially expressed. PLoS Pathog 2 (3) e23.

16. BarthS, PfuhlT, MamianiA, EhsesC, RoemerK, et al. (2008) Epstein-Barr virus-encoded microRNA miR-BART2 down-regulates the viral DNA polymerase BALF5. Nucleic Acids Res 36 (2) 666–75.

17. RileyKJ, RabinowitzGS, YarioTA, LunaJM, DarnellRB, et al. (2012) EBV and human microRNAs co-target oncogenic and apoptotic viral and human genes during latency. EMBO J 31 (9) 2207–21.

18. LungRW, TongJH, SungYM, LeungPS, NgDC, et al. (2009) Modulation of LMP2A expression by a newly identified Epstein-Barr virus-encoded microRNA miR-BART22. Neoplasia 11 (11) 1174–84.

19. XiaT, O'HaraA, AraujoI, BarretoJ, CarvalhoE, et al. (2008) EBV microRNAs in primary lymphomas and targeting of CXCL-11 by ebv-mir-BHRF1-3. Cancer Res 68 (5) 1436–42.

20. NachmaniD, Stern-GinossarN, SaridR, MandelboimO (2009) Diverse herpesvirus microRNAs target the stress-induced immune ligand MICB to escape recognition by natural killer cells. Cell Host Microbe 5 (4) 376–85.

21. HaneklausM, GerlicM, Kurowska-StolarskaM, RaineyAA, PichD, et al. (2012) Cutting edge: miR-223 and EBV miR-BART15 regulate the NLRP3 inflammasome and IL-1β production. J Immunol 189 (8) 3795–9.

22. LeiT, YuenKS, XuR, TsaoSW, ChenH, et al. (2012) Targeting of DICE1 tumor suppressor by Epstein-Barr virus-encoded miR-BART3* microRNA in nasopharyngeal carcinoma. Int J Cancer 133 (1) 79–87.

23. ChoiH, LeeH, KimSR, GhoYS, LeeSK (2013) Epstein-Barr Virus-Encoded MicroRNA BART15-3p Promotes Cell Apoptosis Partially by Targeting BRUCE. J Virol 87 (14) 8135–44.

24. GhoshZ, MallickB, ChakrabartiJ (2009) Cellular versus viral microRNAs in host-virus interaction. Nucleic Acids Res 37 (4) 1035–48.

25. CullenBR (2006) Viruses and microRNAs. Nat Genet 38 Suppl: S25–30.

26. GrassmannR, JeangKT (2008) The roles of microRNAs in mammalian virus infection. Biochim Biophys Acta 1779 (11) 706–11.

27. LoAK, DawsonCW, JinDY, LoKW (2012) The pathological roles of BART miRNAs in nasopharyngeal carcinoma. J Pathol 227 (4) 392–403.

28. MarquitzAR, Raab-TraubN (2012) The role of miRNAs and EBV BARTs in NPC. Semin Cancer Biol 22 (2) 166–72.

29. ChenSJ, ChenGH, ChenYH, LiuCY, ChangKP, et al. (2010) Characterization of Epstein-Barr virus miRNAome in nasopharyngeal carcinoma by deep sequencing. PLoS One 5 (9) pii: e12745 doi: 10.1371/journal.pone.0012745

30. RamakrishnanR, DonahueH, GarciaD, TanJ, ShimizuN, et al. (2011) Epstein-Barr virus BART9 miRNA modulates LMP1 levels and affects growth rate of nasal NK T cell lymphomas. PLoS One 6 (11) e27271.

31. OnderTT, GuptaPB, ManiSA, YangJ, LanderES, et al. (2008) Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res 68 (10) 3645–54.

32. ChenHC, ChenGH, ChenYH, LiaoWL, LiuCY, et al. (2009) MicroRNA deregulation and pathway alterations in nasopharyngeal carcinoma. Br J Cancer 100 (6) 1002–11.

33. MurakamiM, LanK, SubramanianC, RobertsonES (2005) Epstein-Barr virus nuclear antigen 1 interacts with Nm23-H1 in lymphoblastoid cell lines and inhibits its ability to suppress cell migration. J Virol 79 (3) 1559–68.

34. CanelM, SerrelsA, FrameMC, BruntonVG (2013) E-cadherin-integrin crosstalk in cancer invasion and metastasis. J Cell Sci 15;126 (Pt 2) 393–401.

35. ZhengZ, PanJ, ChuB, WongYC, CheungAL, et al. (1999) Downregulation and abnormal expression of E-cadherin and beta-catenin in nasopharyngeal carcinoma: close association with advanced disease stage and lymph node metastasis. Hum Pathol 30 (4) 458–66.

36. LiZ, RenY, LinSX, LiangYJ, LiangHZ (2004) Association of E-cadherin and beta-catenin with metastasis in nasopharyngeal carcinoma. Chin Med J (Engl) 117 (8) 1232–9.

37. LuoW, FangW, LiS, YaoK (2012) Aberrant expression of nuclear vimentin and related epithelial-mesenchymal transition markers in nasopharyngeal carcinoma. Int J Cancer 131 (8) 1863–73.

38. WuCC, ChienKY, TsangNM, ChangKP, HaoSP, et al. (2005) Cancer cell-secreted proteomes as a basis for searching potential tumor markers: nasopharyngeal carcinoma as a model. Proteomics 5 (12) 3173–82.

39. MaL, YoungJ, PrabhalaH, PanE, MestdaghP, et al. (2010) miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis. Nat Cell Biol 12 (3) 247–56.

40. YoshizakiT (2002) Promotion of metastasis in nasopharyngeal carcinoma by Epstein-Barr virus latent membrane protein-1. Histol Histopathol 17 : 845–850.

41. LiuHP, ChenCC, WuCC, HuangYC, LiuSC, et al. (2012) Epstein-Barr virus-encoded LMP1 interacts with FGD4 to activate Cdc42 and thereby promote migration of nasopharyngeal carcinoma cells. PLoS Pathog 8 (5) e1002690.

42. LuJJ, ChenJY, HsuTY, YuWC, SuIJ, et al. (1996) Induction of apoptosis in epithelial cells by Epstein-Barr virus latent membrane protein 1. J Gen Virol 77 (Pt 8) 1883–92.

43. EliopoulosAG, DawsonCW, MosialosG, FloettmannJE, RoweM, et al. (1996) CD40-induced growth inhibition in epithelial cells is mimicked by Epstein-Barr Virus-encoded LMP1: involvement of TRAF3 as a common mediator. Oncogene 13 (10) 2243–54.

44. PathmanathanR, PrasadU, SadlerR, FlynnK, Raab-TraubN (1995) Clonal proliferations of cells infected with Epstein-Barr virus in preinvasive lesions related to nasopharyngeal carcinoma. N Engl J Med 333 (11) 693–8.

45. LanYY, HsiaoJR, ChangKC, ChangJS, ChenCW, et al. (2012) 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 86 (12) 6656–67.

46. FotheringhamJA, CoalsonNE, Raab-TraubN (2012) Epstein-Barr virus latent membrane protein-2A induces ITAM/Syk -⁠ and Akt-dependent epithelial migration through αv-integrin membrane translocation. J Virol 86 (19) 10308–20.

47. KongQL, HuLJ, CaoJY, HuangYJ, XuLH, et al. (2010) Epstein-Barr virus-encoded LMP2A induces an epithelial-mesenchymal transition and increases the number of side population stem-like cancer cells in nasopharyngeal carcinoma. PLoS Pathog 6 (6) e1000940.

48. HuangGW, MoWN, KuangGQ, NongHT, WeiMY, et al. (2001) Expression of p16, nm23-H1, E-cadherin, and CD44 gene products and their significance in nasopharyngeal carcinoma. Laryngoscope 111 (8) 1465–71.

49. KrishnaSM, KattoorJ, BalaramP (2005) Down regulation of adhesion protein E-cadherin in Epstein-Barr virus infected nasopharyngeal carcinomas. Cancer Biomark 1 (6) 271–7.

50. KaoRH, HuangLC, HsuYH (2002) Mapping the methylation pattern by bisulfite genomic sequencing of the E-cadherin promoter CpG island in nasopharyngeal carcinoma. Anticancer Res 22 (6C) 4109–13.

51. TsaoSW, LiuY, WangX, YuenPW, LeungSY, et al. (2003) The association of E-cadherin expression and the methylation status of the E-cadherin gene in nasopharyngeal carcinoma cells. Eur J Cancer 39 (4) 524–31.

52. TsaiCL, LiHP, LuYJ, HsuehC, LiangY, et al. (2006) Activation of DNA methyltransferase 1 by EBV LMP1 Involves c-Jun NH(2)-terminal kinase signaling. Cancer Res 66 (24) 11668–76.

53. HorikawaT, YoshizakiT, KondoS, FurukawaM, KaizakiY, et al. (2011) Epstein-Barr Virus latent membrane protein 1 induces Snail and epithelial-mesenchymal transition in metastatic nasopharyngeal carcinoma. Br J Cancer 104 (7) 1160–7.

54. GregoryPA, BertAG, PatersonEL, BarrySC, TsykinA, et al. (2008) The miR-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol 10 (5) 593–601.

55. AhmadA, AboukameelA, KongD, WangZ, SethiS, et al. (2011) Phosphoglucose isomerase/autocrine motility factor mediates epithelial-mesenchymal transition regulated by miR-200 in breast cancer cells. Cancer Res 71 (9) 3400–9.

56. LiaoSK, PerngYP, ShenYC, ChungPJ, ChangYS, et al. (1998) Chromosomal abnormalities of a new nasopharyngeal carcinoma cell line (NPC-BM1) derived from a bone marrow metastatic lesion. Cancer Genet Cytogenet 103 (1) 52–8.

57. LinCT, WongCI, ChanWY, TzungKW, HoJK, et al. (1990) Establishment and characterization of two nasopharyngeal carcinoma cell lines. Lab Invest 62 (6) 713–24.