Silencing of NUF2 Inhibits Tumor Growth and Induces Apoptosis in Human Hepatocellular Carcinomas

  • Liu, Qiang (Department of Radiology, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University) ;
  • Dai, She-Jiao (Department of Gastroenterology, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University) ;
  • Li, Hong (Department of Gastroenterology, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University) ;
  • Dong, Lei (Department of Gastroenterology, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University) ;
  • Peng, Yu-Ping (Department of Radiology, the Second Affiliated Hospital, Medical School of Xi'an Jiaotong University)
  • 발행 : 2014.11.06


Background: As an important component of the NDC80 kinetochore complex, NUF2 is essential for kinetochore-microtubule attachment and chromosome segregation. Previous studies also suggested its involvement in development of various kinds of human cancers, however, its expression and functions in human hepatocellular carcinoma (HCC) are still unclear. Materials and Methods: In the present study, we aimed to test the hypothesis that NUF2 is aberrant in human HCCs and associated with cell growth. Results: Our results showed significantly elevated expression of NUF2 in human HCC tissues compared to adjacent normal tissues, and high expression of NUF2 in HCC cell lines. Using lentivirus-mediated silencing of NUF2 in HepG2 human HCC cells, we found that NUF2 depletion markedly suppressed proliferation and colony formation capacity in vitro, and dramatically hampered tumor growth of xenografts in vivo. Moreover, NUF2 silencing could induce cell cycle arrest and trigger cell apoptosis. Additionally, altered levels of cell cycle and apoptosis related proteins including cyclin B1, Cdc25A, Cdc2, Bad and Bax were also observed. Conclusions: In conclusion, these results demonstrate that NUF2 plays a critical role in the regulation of HCC cell proliferation and apoptosis, indicating that NUF2 may serve as a potential molecular target for therapeutic approaches.


  1. Bharadwaj R, Yu H (2004). The spindle checkpoint, aneuploidy, and cancer. Oncogene, 23, 2016-27.
  2. Block TM, Mehta AS, Fimmel CJ, et al (2003). Molecular viral oncology of hepatocellular carcinoma. Oncogene, 22, 5093-107.
  3. Boutros R, Lobjois V, Ducommun B (2007). CDC25 phosphatases in cancer cells: key players? Good targets? Nat Rev Cancer, 7, 495-507.
  4. Cahill DP, Lengauer C, Yu J, et al (1998). Mutations of mitotic checkpoint genes in human cancers. Nature, 392, 300-3.
  5. Cheeseman IM, Chappie JS, Wilson-Kubalek EM, et al (2006). The conserved KMN network constitutes the core microtubule-binding site of the kinetochore. Cell, 127, 983-97.
  6. Chen WQ, Zheng RS, Zhang SW (2013). Liver cancer incidence and mortality in China, 2009. Chin J Cancer, 32, 162-9.
  7. Ciferri C, De Luca J, Monzani S, et al (2005). Architecture of the human ndc80-hec1 complex, a critical constituent of the outer kinetochore. J Biol Chem, 280, 29088-95.
  8. DeLuca JG, Dong Y, Hergert P, et al (2005). Hec1 and nuf2 are core components of the kinetochore outer plate essential for organizing microtubule attachment sites. Mol Biol Cell, 16, 519-31.
  9. DeLuca JG, Moree B, Hickey JM, et al (2002). hNuf2 inhibition blocks stable kinetochore-microtubule attachment and induces mitotic cell death in HeLa cells. J Cell Biol, 159, 549-55.
  10. Evans AA, Chen G, Ross EA, et al (2002). Eight-year follow-up of the 90, 000-person Haimen City cohort: I. Hepatocellular carcinoma mortality, risk factors, and gender differences. Cancer Epidemiol Biomarkers Prev, 11, 369-76.
  11. Faivre S, Bouattour M, Raymond E (2011). Novel molecular therapies in hepatocellular carcinoma. Liver Int, 31, 151-60.
  12. Fan JH, Wang JB, Jiang Y, et al (2013). Attributable causes of liver cancer mortality and incidence in China. Asian Pac J Cancer Prev, 14, 7251-56.
  13. Foley EA, Kapoor TM (2013). Microtubule attachment and spindle assembly checkpoint signalling at the kinetochore. Nat Rev Mol Cell Biol, 14, 25-37.
  14. Hanahan D, Weinberg RA (2011). Hallmarks of cancer: the next generation. Cell, 144, 646-74.
  15. Harao M, Hirata S, Irie A, et al (2008). HLA-A2-restricted CTL epitopes of a novel lung cancer-associated cancer testis antigen, cell division cycle associated 1, can induce tumorreactive CTL. Int J Cancer, 123, 2616-25.
  16. Hayama S, Daigo Y, Kato T, et al (2006). Activation of CDCA1-KNTC2, members of centromere protein complex, involved in pulmonary carcinogenesis. Cancer Res, 66, 10339-48.
  17. Jemal A, Bray F, Center MM, et al (2011). Global cancer statistics. CA Cancer J Clin, 61, 69-90.
  18. Kaneko N, Miura K, Gu Z, et al (2009). siRNA-mediated knockdown against CDCA1 and KNTC2, both frequently overexpressed in colorectal and gastric cancers, suppresses cell proliferation and induces apoptosis. Biochem Biophys Res Commun, 390, 1235-40.
  19. Kim HS, Park KH, Kim SA, et al (2005). Frequent mutations of human Mad2, but not Bub1, in gastric cancers cause defective mitotic spindle checkpoint. Mutat Res, 578, 187-201.
  20. Liu D, Ding X, Du J, et al (2007). Human NUF2 interacts with centromere-associated protein E and is essential for a stable spindle microtubule-kinetochore attachment. J Biol Chem, 282, 21415-24.
  21. Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2 (-Delta Delta C (T)) Method. Methods, 25, 402-8.
  22. Maity A, McKenna WG, Muschel RJ (1995). Evidence for post-transcriptional regulation of cyclin B1 mRNA in the cell cycle and following irradiation in HeLa cells. EMBO J, 14, 603-9.
  23. McCleland ML, Gardner RD, Kallio MJ, et al (2003). The highly conserved Ndc80 complex is required for kinetochore assembly, chromosome congression, and spindle checkpoint activity. Genes Dev, 17, 101-14.
  24. Nabetani A, Koujin T, Tsutsumi C, et al (2001). A conserved protein, Nuf2, is implicated in connecting the centromere to the spindle during chromosome segregation: a link between the kinetochore function and the spindle checkpoint. Chromosoma, 110, 322-34.
  25. Ohnuma S, Miura K, Horii A, et al (2009). Cancer-associated splicing variants of the CDCA1 and MSMB genes expressed in cancer cell lines and surgically resected gastric cancer tissues. Surgery, 145, 57-68.
  26. Rajagopalan H, Lengauer C (2004). Aneuploidy and cancer. Nature, 432, 338-41.
  27. Santaguida S, Musacchio A (2009). The life and miracles of kinetochores. EMBO J, 28, 2511-31.
  28. Santamaria D, Barriere C, Cerqueira A, et al (2007) Cdk1 is sufficient to drive the mammalian cell cycle. Nature, 448, 811-5.
  29. Sethi G, Pathak HB, Zhang H, et al (2012). An RNA interference lethality screen of the human druggable genome to identify molecular vulnerabilities in epithelial ovarian cancer. PLoS One, 7, 47086.
  30. Thorgeirsson SS, Grisham JW (2002). Molecular pathogenesis of human hepatocellular carcinoma. Nat Genet, 31, 339-46.
  31. Tomonaga T, Matsushita K, Ishibashi M, et al (2005). Centromere protein H is up-regulated in primary human colorectal cancer and its overexpression induces aneuploidy. Cancer Res, 65, 4683-9.
  32. Tomonaga T, Matsushita K, Yamaguchi S, et al (2003). Overexpression and mistargeting of centromere protein-A in human primary colorectal cancer. Cancer Res, 63, 3511-6.
  33. Walker MG (2001). Drug target discovery by gene expression analysis: cell cycle genes. Curr Cancer Drug Targets, 1, 73-83.
  34. Wigge PA, Kilmartin JV (2001). The Ndc80p complex from Saccharomyces cerevisiae contains conserved centromere components and has a function in chromosome segregation. J Cell Biol, 152, 349-60.
  35. Yuen KW, Montpetit B, Hieter P (2005). The kinetochore and cancer: what's the connection? Curr Opin Cell Biol, 17, 576-82.

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