Tumor Biology

, Volume 35, Issue 2, pp 1503–1510 | Cite as

miR-34 is associated with poor prognosis of patients with gallbladder cancer through regulating telomere length in tumor stem cells

  • Ke Jin
  • Yonghua Xiang
  • Jing Tang
  • Guangchun Wu
  • Junwei Li
  • Huaichun Xiao
  • Chunwang Li
  • Yuxiang Chen
  • Jingfeng ZhaoEmail author
Research Article


miR-34a has been identified as a tumor suppressor in several tumors, but its involvement in gallbladder cancer (GBC) has not been reported. In this study, the miR-34a level and telomere length were measured in 77 gallbladder adenocarcinomas and 36 peritumoral tissues by real-time PCR. Forced miR-34a expression was established by an adenovirus carrying a miR-34a expression cassette. The colony-forming ability of isolated CD44+CD133+ GBC tumor stem-like cells was measured by matrigel colony assay. The xenograft tumor models were established by inoculating nude mice with CD44+CD133+cells. Results showed that significantly lower miR-34a expression and longer telomere length were observed in gallbladder adenocarcinoma tissues, which correlated with poor prognosis of GBC patients. Forced overexpression of miR-34a inhibited the colony-forming ability of CD44+CD133+ GBC tumor stem-like cells in vitro and xenograft tumor growth in vivo. Injection of Ad-miR-34a downregulated PNUTS expression and reduced telomere length in xenograft GBC tumor cells. In conclusion, miR-34a is a tumor suppressor in gallbladder cancer. Both low miR-34a expression and long telomere length are markers for poor prognosis of patients with gallbladder adenocarcinoma. Our study also suggests that the miR-34a gene could be a target for targeting therapy of GBC.


Gallbladder cancer miR-34a Telomere length Prognosis 



This study was supported by the National Hi-tech Program (863 Project) of China (No. 2007AA021804, 2007AA021809).

Conflicts of interest



  1. 1.
    Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, et al. Cancer statistics 2008. CA Cancer J Clin. 2008;58:71–96.PubMedCrossRefGoogle Scholar
  2. 2.
    Jayaraman S, Jarnagin WR. Management of gallbladder cancer. Gastroenterol Clin North Am. 2010;39:331–42.PubMedCrossRefGoogle Scholar
  3. 3.
    Hawkins WG, DeMatteo RP, Jarnagin WR, Ben-Porat L, Blumgart LH, Fong Y. Jaundice predicts advanced disease and early mortality in patients with gallbladder cancer. Ann Surg Oncol. 2004;11:310–5.PubMedCrossRefGoogle Scholar
  4. 4.
    Ootani T, Shirai Y, Tsukada K, Muto T. Relationship between gallbladder carcinoma and the segmental type of adenomyomatosis of the gallbladder. Cancer. 1992;69:2647–52.PubMedCrossRefGoogle Scholar
  5. 5.
    Roa JC, Tapia O, Cakir A, Basturk O, Dursun N, Akdemir D, et al. Squamous cell and adenosquamous carcinomas of the gallbladder: clinicopathological analysis of 34 cases identified in 606 carcinomas. Mod Pathol. 2011;24:1069–78.PubMedCrossRefGoogle Scholar
  6. 6.
    Park SB, Kim YH, Rho HL, Chae GB, Hong SK. Primary carcinosarcoma of the gallbladder. J Korean Surg Soc. 2012;82:54–8.PubMedCentralPubMedCrossRefGoogle Scholar
  7. 7.
    Valencia-Sanchez MA, Liu J, Hannon GJ, Parker R. Control of translation and mRNA degradation by miRNAs and siRNAs. Genes Dev. 2006;20:515–24.PubMedCrossRefGoogle Scholar
  8. 8.
    Bagga S, Pasquinelli AE. Identification and analysis of microRNAs. Genet Eng (N Y). 2006;27:1–20.CrossRefGoogle Scholar
  9. 9.
    Xiong J, Du Q, Liang Z. Tumor-suppressive microRNA-22 inhibits the transcription of E-box-containing c-Myc target genes by silencing c-Myc binding protein. Oncogene. 2010;29:4980–8.PubMedCrossRefGoogle Scholar
  10. 10.
    Raver-Shapira N, Marciano E, Meiri E, Spector Y, Rosenfeld N, Moskovits N, et al. Transcriptional activation of miR-34a contributes to p53-mediated apoptosis. Mol Cell. 2007;26:731–43.PubMedCrossRefGoogle Scholar
  11. 11.
    Tarasov V, Jung P, Verdoodt B, Lodygin D, Epanchintsev A, Menssen A, et al. Differential regulation of microRNAs by p53 revealed by massively parallel sequencing: miR-34a is a p53 target that induces apoptosis and G1-arrest. Cell Cycle. 2007;6:1586–93.PubMedCrossRefGoogle Scholar
  12. 12.
    Corney DC, Flesken-Nikitin A, Godwin AK, Wang W, Nikitin AY. MicroRNA-34b and microRNA-34c are targets of p53 and cooperate in control of cell proliferation and adhesion-independent growth. Cancer Res. 2007;67:8433–8.PubMedCrossRefGoogle Scholar
  13. 13.
    Bommer GT, Gerin I, Feng Y, Kaczorowski AJ, Kuick R, Love RE, et al. p53-mediated activation of miRNA34 candidate tumor-suppressor genes. Curr Biol. 2007;17:1298–307.PubMedCrossRefGoogle Scholar
  14. 14.
    Chang TC, Wentzel EA, Kent OA, Ramachandran K, Mullendore M, Lee KH, et al. Transactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosis. Mol Cell. 2007;26:745–52.PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Lodygin D, Tarasov V, Epanchintsev A, Berking C, Knyazeva T, Korner H, et al. Inactivation of miR-34a by aberrant CpG methylation in multiple types of cancer. Cell Cycle. 2008;7:2591–600.PubMedCrossRefGoogle Scholar
  16. 16.
    Boon RA, Iekushi K, Lechner S, Seeger T, Fischer A, Heydt S, et al. MicroRNA-34a regulates cardiac ageing and function. Nature. 2013;495:107–10.PubMedCrossRefGoogle Scholar
  17. 17.
    Sahin E, DePinho RA. Axis of ageing: telomeres, p53 and mitochondria. Nat Rev Mol Cell Biol. 2012;13:397–404.PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Cawthon RM. Telomere measurement by quantitative PCR. Nucleic Acids Res. 2002;30:e47.PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Hu Y, Ou Y, Wu K, Chen Y, Sun W. miR-143 inhibits the metastasis of pancreatic cancer and an associated signaling pathway. Tumour Biol. 2012;33:1863–70.PubMedCrossRefGoogle Scholar
  20. 20.
    Zhang X, Kon T, Wang H, Li F, Huang Q, Rabbani ZN, et al. Enhancement of hypoxia-induced tumor cell death in vitro and radiation therapy in vivo by use of small interfering RNA targeted to hypoxia-inducible factor-1alpha. Cancer Res. 2004;64:8139–42.PubMedCrossRefGoogle Scholar
  21. 21.
    Ogawa T, Saiki Y, Shiga K, Chen N, Fukushige S, Sunamura M, et al. miR-34a is downregulated in cis-diamminedichloroplatinum treated sinonasal squamous cell carcinoma patients with poor prognosis. Cancer Sci. 2012;103:1737–43.PubMedCrossRefGoogle Scholar
  22. 22.
    Gallardo E, Navarro A, Viñolas N, Marrades RM, Diaz T, Gel B, et al. miR-34a as a prognostic marker of relapse in surgically resected non-small-cell lung cancer. Carcinogenesis. 2009;30:1903–9.PubMedCrossRefGoogle Scholar
  23. 23.
    Hermeking H. The miR-34 family in cancer and apoptosis. Cell Death Differ. 2010;17:193–9.PubMedCrossRefGoogle Scholar
  24. 24.
    Eisenberg DTA. An evolutionaty review of human telomere biology: The thirty telomere hypothesis and notes on potential adaptive paternal effects. Am J Hum Biol. 2011;23:149–67.PubMedCrossRefGoogle Scholar
  25. 25.
    Svenson U, Roos G. Telomere length as a biological marker in malignancy. Biochim Biophys Acta. 2009;1792:317–23.PubMedCrossRefGoogle Scholar
  26. 26.
    Bisofti M, Heaphy CM, Griffith JK. Telomeres: Prognostic markers for solid tumors. Int J Cancer. 2006;119:2255–60.CrossRefGoogle Scholar
  27. 27.
    Avigad S, Naumov I, Ohali A, Jeison M, Berco GH, Mardoukh J, et al. Short telomeres: a novel potential predictor of relapse in Ewing sarcoma. Clin Cancer Res. 2007;13:5777–83.PubMedCrossRefGoogle Scholar
  28. 28.
    Shirotani Y, Hiyama K, Ishioka S, Inyaku K, Awaya Y, Yonehara S, et al. Alteration in length of telomeric repeats in lung cancer. Lung Cancer. 1994;11:29–41.PubMedCrossRefGoogle Scholar
  29. 29.
    Hirashima T, Komiya T, Nitta T, Takada Y, Kobayashi M, Masuda N, et al. Prognostic significance of telomeric repeat length alterations in pathological stage I–IIIA non-small cell lung cancer. Anticancer Res. 2000;20:2181–7.PubMedGoogle Scholar
  30. 30.
    Frías C, García-Aranda C, De Juan C, Morán A, Ortega P, Gómez A, et al. Telomere shortening is associated with poor prognosis and telomerase activity correlates withDNArepair impairment in non-small cell lung cancer. Lung Cancer. 2008;60:416–25.PubMedCrossRefGoogle Scholar
  31. 31.
    Shi C, Tian R, Wang M, Wang X, Jiang J, Zhang Z, et al. CD44 + CD133+ population exhibits cancer stem cell-like characteristics in human gallbladder carcinoma. Cancer Biol Ther. 2010;11:1182–90.CrossRefGoogle Scholar
  32. 32.
    Kim H, Lee OH, Xin H, Chen LY, Qin J, Chae HK, et al. TRF2 functions as a protein hub and regulates telomere maintenance by recognizing specific peptide motifs. Nat Struct Mol Biol. 2009;16:372–9.PubMedCrossRefGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2013

Authors and Affiliations

  • Ke Jin
    • 1
  • Yonghua Xiang
    • 1
  • Jing Tang
    • 1
  • Guangchun Wu
    • 1
  • Junwei Li
    • 1
  • Huaichun Xiao
    • 1
  • Chunwang Li
    • 1
  • Yuxiang Chen
    • 2
  • Jingfeng Zhao
    • 2
    Email author
  1. 1.Department of RadiologyHunan Children’s HospitalChangshaChina
  2. 2.Hepatobiliary & Enteric Surgery Research Center, Xiangya HospitalCentral South UniversityChangshaChina

Personalised recommendations