Tumor Biology

, Volume 37, Issue 12, pp 15925–15936 | Cite as

E2F-1 promotes DAPK2-induced anti-tumor immunity of gastric cancer cells by targeting miR-34a

  • Lin-Hai Yan
  • Zhi-Ning Chen
  • Li Li
  • Jia Chen
  • Xian-Wei Mo
  • Yu-Zhou Qin
  • Wen-E Wei
  • Hai-Quan Qin
  • Yuan Lin
  • Jian-Si Chen
Original Article


Activation of the transcription factor E2F-1 gene is a negative event in dendritic cell (DC) maturation process. Down-regulation of E2F1 causes immaturity of DC thereby stopping antigen production which in turn leads to inhibition of immune responses. E2F-1-free stimulates the NF-kB signaling pathway, leading to activation of monocytes and several other transcription factor genes. In the study, we report that down-regulation of E2F-1 in DCs promote anti-tumor immune response in gastric cancer (GC) cells through a novel mechanism. DCs were isolated from peripheral blood mononuclear cells. E2F-1 small interfering RNA (E2F-1-shRNA) induced down-regulation of E2F-1 mRNA and protein expression in DCs. Furthermore, we identified the E2F-1-shRNA targeted the CD80, CD83, CD86, and MHC II molecules, promoted their expression, and induced T lymphocytes proliferation activity and up-regulation of IFN-Ī³ production and GC cell killing effect, which significantly correlated with the cytotoxic T lymphocytes activated by E2F-1-shRNA DCs. The higher expression of miR-34a was found which was significantly correlated with the DC enhancing anti-tumor immunity against gastric cancer cell, and miR-34a potently targeted DAPK2 and Sp1, both of which were involved in the deactivation of E2F-1. Moreover, in E2F-1-DC-down-regulation in mice, GC transplantation tumors displayed down-regulation of Sp1, DAPK2, Caspase3, and Caspase7 and progressed to anti-tumor immunity. Collectively, our data uncover an E2F-1-mediated mechanism for the control of DC anti-tumor immunity via miR-34a-dependent down-regulation of E2F-1 expression and suggest its contribution to GC immunotherapy.


Transcription factor E2F-1 Dendritic cell MicroRNA Gastric cancer cells Immunotherapy 



We acknowledge financial support from the National Science Foundation for Young Scholars of China (No. 81502120), the Science Foundation for Young Scholars of Guangxi Medical University (No. GXMUYSF201404), and the Scientific Research and Technological Foundation of Guangxi (No.1140003A-35). Also, we express our gratitude to Ms. Shwetha Manoj, who made extraordinary help in paper editing.

Compliance with ethical standards

The study design was approved by the Guangxi medical university ethic review board obeying the Helsinki Declaration. All participants have signed informed consent forms and agreed the treatment.

The mice were normatively managed by SPF laboratory in the Animal Central of Guangxi Medical University, following the rules of ethical regulations for medical experimental animal care.

Conflicts of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.

Supplementary material

13277_2016_5446_MOESM1_ESM.docx (171 kb)
Supplementary figure 1 Down-regulation of E2F-1 decreased miR-23a, miR-27a, miR-126, miR-133a, and miR-148b, and increased miR-34a expression. The determination of miRNA expression levels of miR-23a, miR-27a, miR-126, miR-133a, miR-148b, and miR-34a through semiquantitative reverse-transcriptase polymerase chain reaction. (DOCX 170Ā kb)
13277_2016_5446_MOESM2_ESM.docx (271 kb)
Supplementary figure 2 FACS analysis of peripheral blood monocyte-derived DCs. PBMCs cultured in the presence of GM-CSF (50Ā ng/mL), IL-4 (50Ā ng/mL), and LPS (100Ā ng/mL). Maturation of DCs is marked by the increased levels of CD83 and CD86. There is also a marked increase in the expression of HLA-DR. The numbers in the boxes indicate the mean fluorescence intensity. (DOCX 270Ā kb)


  1. 1.
    Zhou M, Wang H, Zhu J, Chen W, Wang L, Liu S, et al. Cause-specific mortality for 240 causes in China during 1990-2013: a systematic subnational analysis for the global burden of disease study 2013. Lancet. 2015;23:551–6.Google Scholar
  2. 2.
    van Beek JJ, Wimmers F, Hato SV, de Vries IJ, Skold AE. Dendritic cell cross talk with innate and innate-like effector cells in antitumor immunity: implications for DC vaccination. Crit Rev Immunol. 2014;34:517–36.CrossRefPubMedGoogle Scholar
  3. 3.
    N'Diaye M, Warnecke A, Flytzani S, Abdelmagid N, Ruhrmann S, Olsson T, Jagodic M, et al. Rat bone marrow-derived dendritic cells generated with GM-CSF/IL-4 or FLT3L exhibit distinct phenotypical and functional characteristics. J Leukoc Biol. 2015;29:e914.Google Scholar
  4. 4.
    Kantoff PW, Higano CS, Shore ND, Berger ER, Small EJ, Penson DF, et al. Sipuleucel-T immunotherapy for castration-resistant prostate cancer. N Engl J Med. 2010;363:411–22.CrossRefPubMedGoogle Scholar
  5. 5.
    Yin X, Johns SC, Kim D, Mikulski Z, Salanga CL, Handel TM, et al. Lymphatic specific disruption in the fine structure of heparan sulfate inhibits dendritic cell traffic and functional T cell responses in the lymph node. J Immunol. 2014;192:2133–42.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Fang F, Wang Y, Li R, Zhao Y, Guo Y, Jiang M, Sun J, et al. Transcription factor E2F1 suppresses dendritic cell maturation. J Immunol. 2010;184:6084–91.CrossRefPubMedGoogle Scholar
  7. 7.
    Desrichard A, Snyder A, Chan TA. Cancer neoantigens and applications for immunotherapy. Clin Cancer Res. 2015;22:1–6.Google Scholar
  8. 8.
    Swiecki M, Colonna M. The multifaceted biology of plasmacytoid dendritic cells. Nat Rev Immunol. 2015;15:471–85.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Yan LH, Wei WY, Cao WL, Zhang XS, Xie YB, Xiao Q. Overexpression of E2F1 in human gastric carcinoma is involved in anti-cancer drug resistance. BMC Cancer. 2014;14:904.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Yamasaki L, Jacks T, Bronson R, Goillot E, Harlow E, Dyson NJ. Tumor induction and tissue atrophy in mice lacking E2F-1. Cell. 1996;85:537–48.CrossRefPubMedGoogle Scholar
  11. 11.
    Field SJ, Tsai FY, Kuo F, Zubiaga AM, Kaelin Jr WG, et al. E2F-1 functions in mice to promote apoptosis and suppress proliferation. Cell. 1996;85:549–61.CrossRefPubMedGoogle Scholar
  12. 12.
    Zhu JW, Field SJ, Gore L, Thompson M, Yang H, Fujiwara Y, et al. E2F1 and E2F2 determine thresholds for antigen-induced T-cell proliferation and suppress tumorigenesis. Mol Cell Biol. 2001;21:8547–64.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Yan LH, Wang XT, Yang J, Kong FB, Lian C, Wei WY, et al. Reversal of multidrug resistance in gastric cancer cells by E2F-1 downregulation in vitro and in vivo, J. Cell Biochem. 2014;115:34–41.CrossRefGoogle Scholar
  14. 14.
    Kuang Y, Weng X, Liu X, Zhu H, Chen Z, Jiang B, et al. Anti-tumor immune response induced by dendritic cells transduced with truncated PSMA IRES 4-1BBL recombinant adenoviruses. Cancer Lett. 2010;293:254–62.CrossRefPubMedGoogle Scholar
  15. 15.
    Aranda F, Vacchelli E, Eggermont A, Galon J, Sautes-Fridman C, Tartour E, Zitvogel L, Kroemer G, et al. Trial watch: peptide vaccines in cancer therapy. Oncoimmunology. 2013;2:e26621.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Zheng C, Ren Z, Wang H, Zhang W, Kalvakolanu DV, Tian Z, et al. E2F1 induces tumor cell survival via nuclear factor-kappaB-dependent induction of EGR1 transcription in prostate cancer cells. Cancer Res. 2009;69:2324–31.CrossRefPubMedGoogle Scholar
  17. 17.
    Britschgi A, Trinh E, Rizzi M, Jenal M, Ress A, Tobler A, et al. DAPK2 is a novel E2F1/KLF6 target gene involved in their proapoptotic function. Oncogene. 2008;27:5706–16.CrossRefPubMedGoogle Scholar
  18. 18.
    Nencioni A, Grunebach F, Schmidt SM, Muller MR, Boy D, Patrone F, et al. The use of dendritic cells in cancer immunotherapy. Crit Rev Oncol Hematol. 2008;65:191–9.CrossRefPubMedGoogle Scholar
  19. 19.
    Aslanidi GV, Rivers AE, Ortiz L, Govindasamy L, Ling C, Jayandharan GR, et al. High-efficiency transduction of human monocyte-derived dendritic cells by capsid-modified recombinant AAV2 vectors. Vaccine. 2012;30:3908–17.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Wang B, Ma A, Zhang L, Jin WL, Qian Y, Xu G, et al. POH1 deubiquitylates and stabilizes E2F1 to promote tumour formation. Nat Commun. 2015;6:8704.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    DeRyckere D, DeGregori J. E2F1 and E2F2 are differentially required for homeostasis-driven and antigen-induced T cell proliferation in vivo. J Immunol. 2005;175:647–55.CrossRefPubMedGoogle Scholar
  22. 22.
    Li M, Zhang X, Zheng X, Lian D, Zhang ZX, Ge W, et al. Immune modulation and tolerance induction by RelB-silenced dendritic cells through RNA interference. J Immunol. 2007;178:5480–7.CrossRefPubMedGoogle Scholar
  23. 23.
    Lind EF, Ahonen CL, Wasiuk A, Kosaka Y, Becher B, Bennett KA, et al. Dendritic cells require the NF-kappaB2 pathway for cross-presentation of soluble antigens. J Immunol. 2008;181:354–63.CrossRefPubMedGoogle Scholar
  24. 24.
    Mann J, Oakley F, Johnson PW, Mann DA. CD40 induces interleukin-6 gene transcription in dendritic cells: regulation by TRAF2, AP-1, NF-kappa B, AND CBF1. J Biol Chem. 2002;277:17125–38.CrossRefPubMedGoogle Scholar
  25. 25.
    Jafarinejad-Farsangi S, Farazmand A, Mahmoudi M, Gharibdoost F, Karimizadeh E, Noorbakhsh F, et al. MicroRNA-29a induces apoptosis via increasing the Bax:Bcl-2 ratio in dermal fibroblasts of patients with systemic sclerosis. Autoimmunity. 2015;48:369–78.CrossRefPubMedGoogle Scholar
  26. 26.
    Welch C, Chen Y, Stallings RL. MicroRNA-34a functions as a potential tumor suppressor by inducing apoptosis in neuroblastoma cells. Oncogene. 2007;26:5017–22.CrossRefPubMedGoogle Scholar
  27. 27.
    Xu X, Chen W, Miao R, Zhou Y, Wang Z, Zhang L, et al. miR-34a induces cellular senescence via modulation of telomerase activity in human hepatocellular carcinoma by targeting FoxM1/c-Myc pathway. Oncotarget. 2015;6:3988–4004.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Tryndyak VP, Ross SA, Beland FA, Pogribny IP. Down-regulation of the microRNAs miR-34a, miR-127, and miR-200b in rat liver during hepatocarcinogenesis induced by a methyl-deficient diet. Mol Carcinog. 2009;48:479–87.CrossRefPubMedGoogle Scholar
  29. 29.
    Kumamoto K, Spillare EA, Fujita K, Horikawa I, Yamashita T, Appella E, et al. Nutlin-3a activates p53 to both down-regulate inhibitor of growth 2 and up-regulate mir-34a, mir-34b, and mir-34c expression, and induce senescence. Cancer Res. 2008;68:3193–203.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Christoffersen NR, Shalgi R, Frankel LB, Leucci E, Lees M, Klausen M, et al. p53-independent upregulation of miR-34a during oncogene-induced senescence represses MYC. Cell Death Differ. 2010;17:236–45.CrossRefPubMedGoogle Scholar
  31. 31.
    Bai XY, Ma Y, Ding R, Fu B, Shi S, Chen XM. miR-335 and miR-34a promote renal senescence by suppressing mitochondrial antioxidative enzymes. J Am Soc Nephrol. 2011;22:1252–61.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Lin-Hai Yan
    • 1
  • Zhi-Ning Chen
    • 2
  • Li Li
    • 3
  • Jia Chen
    • 4
  • Xian-Wei Mo
    • 1
  • Yu-Zhou Qin
    • 1
  • Wen-E Wei
    • 5
  • Hai-Quan Qin
    • 1
  • Yuan Lin
    • 1
  • Jian-Si Chen
    • 1
  1. 1.Department of Gastrointestinal SurgeryAffiliated Tumor Hospital of Guangxi Medical UniversityNanningChina
  2. 2.Department of PathologyAffiliated Tumor Hospital of Guangxi Medical UniversityNanningChina
  3. 3.Department of PharmacyThe People Hospital of Guangxi Zhuang Autonomous RegionNanningChina
  4. 4.Department of Medical Image CenterAffiliated Tumor Hospital of Guangxi Medical UniversityNanningChina
  5. 5.Department of ResearchAffiliated Tumor Hospital of Guangxi Medical UniversityNanningChina

Personalised recommendations