Skip to main content

Advertisement

SpringerLink
Log in

Tumor suppressor miR-145 reverses drug resistance by directly targeting DNA damage-related gene RAD18 in colorectal cancer

  • Research Article
  • Published:
Tumor Biology

Abstract

Colorectal cancer (CRC) is one of the most common cancers worldwide. Although chemotherapy is used as a palliative treatment, ultimately, nearly all patients develop drug resistance. Therefore, the cell-inherent DNA repair pathway must reverse the DNA-damaging effect of cytotoxic drugs that mediates therapeutic resistance to chemotherapy. RAD18, a DNA damage-activated E3 ubiquitin ligase, is known to play a critical role in DNA damage repair in cancer cells. Here, we show that RAD18 is highly expressed in human 5-fluorouracil (5-FU)-resistant cancer cells after 5-FU treatment. In addition, RAD18 increases in CRC cells could induce DNA damage repair, suggesting that RAD18 might be a possible target for overcoming drug resistance. Moreover, the expression of tumor suppressor microRNA-145 (miR-145) was negatively correlated with RAD18 expression in CRC tissues of 140 patients. Using luciferase reporters carrying the 3′-untranslated region of RAD18 combined with Western blotting, we identified RAD18 as a direct target of miR-145. Also of interest, suppression of RAD18 by miR-145 enhanced DNA damage in CRC cells after 5-FU treatment. Finally, the 5-FU-resistant cancer cells could be selectively ablated by treatment with miR-145. Taken together, these results suggest that miR-145 can act as an RAD18 inhibitor and contribute as an important factor in reversing drug resistance after chemotherapy.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Weitz J, Koch M, Debus J, Hohler T, Galle PR, Buchler MW. Colorectal cancer. Lancet. 2005;365(9454):153–65.

    Article  PubMed  Google Scholar 

  2. Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer statistics, 2009. CA Cancer J Clin. 2009;59(4):225–49.

    Article  PubMed  Google Scholar 

  3. Johnston PG, Kaye S. Capecitabine: a novel agent for the treatment of solid tumors. Anticancer Drugs. 2001;12(8):639–46.

    Article  CAS  PubMed  Google Scholar 

  4. Giacchetti S, Perpoint B, Zidani R, Le BN, Faggiuolo R, Focan C, et al. Phase III multicenter randomized trial of oxaliplatin added to chronomodulated fluorouracil-leucovorin as first-line treatment of metastatic colorectal cancer. J Clin Oncol. 2000;18(1):136–47.

    Article  CAS  PubMed  Google Scholar 

  5. Lord CJ, Ashworth A. The DNA damage response and cancer therapy. Nature. 2012;481(7381):287–94.

    Article  CAS  PubMed  Google Scholar 

  6. Lawrence CW, Christensen R. UV mutagenesis in radiation-sensitive strains of yeast. Genetics. 1976;82(2):207–32.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Bailly V, Lamb J, Sung P, Prakash S, Prakash L. Specific complex formation between yeast RAD6 and RAD18 proteins: a potential mechanism for targeting RAD6 ubiquitin-conjugating activity to DNA damage sites. Genes Dev. 1994;8(7):811–20.

    Article  CAS  PubMed  Google Scholar 

  8. Xin H, Lin W, Sumanasekera W, Zhang Y, Wu X, Wang Z. The human RAD18 gene product interacts with HHR6A and HHR6B. Nucleic Acids Res. 2000;28(14):2847–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Tateishi S, Niwa H, Miyazaki J, Fujimoto S, Inoue H, Yamaizumi M. Enhanced genomic instability and defective postreplication repair in RAD18 knockout mouse embryonic stem cells. Mol Cell Biol. 2003;23(2):474–81.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Wold MS. Replication protein A: a heterotrimeric, single-stranded DNA-binding protein required for eukaryotic DNA metabolism. Annu Rev Biochem. 1997;66:61–92.

    Article  CAS  PubMed  Google Scholar 

  11. Chiyomaru T, Enokida H, Tatarano S, Kawahara K, Uchida Y, Nishiyama K, et al. miR-145 and miR-133a function as tumour suppressors and directly regulate FSCN1 expression in bladder cancer. Br J Cancer. 2010;102(5):883–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Gregersen LH, Jacobsen AB, Frankel LB, Wen J, Krogh A, Lund AH. MicroRNA-145 targets YES and STAT1 in colon cancer cells. PLOS ONE. 2010;5(1):e8836.

    Article  PubMed  PubMed Central  Google Scholar 

  13. Wang S, Bian C, Yang Z, Bo Y, Li J, Zeng L, et al. miR-145 inhibits breast cancer cell growth through RTKN. Int J Oncol. 2009;34(5):1461–6.

    CAS  PubMed  Google Scholar 

  14. Xu Q, Liu LZ, Qian X, Chen Q, Jiang Y, Li D, et al. MiR-145 directly targets p70S6K1 in cancer cells to inhibit tumor growth and angiogenesis. Nucleic Acids Res. 2012;40(2):761–74.

    Article  CAS  PubMed  Google Scholar 

  15. Nohata N, Sone Y, Hanazawa T, Fuse M, Kikkawa N, Yoshino H, et al. miR-1 as a tumor suppressive microRNA targeting TAGLN2 in head and neck squamous cell carcinoma. Oncotarget. 2011;2(1-2):29–42.

    PubMed  PubMed Central  Google Scholar 

  16. Qu C, Liang Z, Huang J, Zhao R, Su C, Wang S, et al. MiR-205 determines the radioresistance of human nasopharyngeal carcinoma by directly targeting PTEN. Cell Cycle. 2012;11(4):785–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ostenfeld MS, Bramsen JB, Lamy P, Villadsen SB, Fristrup N, Sorensen KD, et al. miR-145 induces caspase-dependent and -independent cell death in urothelial cancer cell lines with targeting of an expression signature present in Ta bladder tumors. Oncogene. 2010;29(7):1073–84.

    Article  CAS  PubMed  Google Scholar 

  18. Lai VK, Ashraf M, Jiang S, Haider K. MicroRNA-143 is a critical regulator of cell cycle activity in stem cells with co-overexpression of Akt and angiopoietin-1 via transcriptional regulation of Erk5/cyclin D1 signaling. Cell Cycle. 2012;11(4):767–77.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Cirera-Salinas D, Pauta M, Allen RM, Salerno AG, Ramirez CM, Chamorro-Jorganes A, et al. Mir-33 regulates cell proliferation and cell cycle progression. Cell Cycle. 2012;11(5):922–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Zou C, Xu Q, Mao F, Li D, Bian C, Liu LZ, et al. MiR-145 inhibits tumor angiogenesis and growth by N-RAS and VEGF. Cell Cycle. 2012;11(11):2137–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Xu N, Papagiannakopoulos T, Pan G, Thomson JA, Kosik KS. MicroRNA-145 regulates OCT4, SOX2, and KLF4 and represses pluripotency in human embryonic stem cells. Cell. 2009;137(4):647–58.

    Article  CAS  PubMed  Google Scholar 

  22. Grimson A, Farh KK, Johnston WK, Garrett-Engele P, Lim LP, Bartel DP. MicroRNA targeting specificity in mammals: determinants beyond seed pairing. Mol Cell. 2007;27(1):91–105.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Yokomine K, Senju S, Nakatsura T, Irie A, Hayashida Y, Ikuta Y, et al. The forkhead box M1 transcription factor as a candidate of target for anti-cancer immunotherapy. Int J Cancer. 2010;126(9):2153–63.

    CAS  PubMed  Google Scholar 

  24. Wang WJ, Wu SP, Liu JB, Shi YS, Huang X, Zhang QB, et al. MYC regulation of CHK1 and CHK2 promotes radioresistance in a stem cell-like population of nasopharyngeal carcinoma cells. Cancer Res. 2013;73(3):1219–31.

    Article  CAS  PubMed  Google Scholar 

  25. Sachdeva M, Zhu S, Wu F, Wu H, Walia V, Kumar S, et al. p53 represses c-Myc through induction of the tumor suppressor miR-145. Proc Natl Acad Sci U S A. 2009;106(9):3207–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Sachdeva M, Mo YY. MicroRNA-145 suppresses cell invasion and metastasis by directly targeting mucin 1. Cancer Res. 2010;70(1):378–87.

    Article  CAS  PubMed  Google Scholar 

  27. Sachdeva M, Zhu S, Wu F, Wu H, Walia V, Kumar S, et al. p53 represses c-Myc through induction of the tumor suppressor miR-145. Proc Natl Acad Sci U S A. 2009;106(9):3207–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Watanabe K, Iwabuchi K, Sun J, Tsuji Y, Tani T, Tokunaga K, et al. RAD18 promotes DNA double-strand break repair during G1 phase through chromatin retention of 53BP1. Nucleic Acids Res. 2009;37(7):2176–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Bouwman P, Jonkers J. The effects of deregulated DNA damage signalling on cancer chemotherapy response and resistance. Nat Rev Cancer. 2012;12(9):587–98.

    Article  CAS  PubMed  Google Scholar 

  30. Davies AA, Neiss A, Ulrich HD. Ubiquitylation of the 9-1-1 checkpoint clamp is independent of rad6-rad18 and DNA damage. Cell. 2010;141(6):1080–7.

    Article  CAS  PubMed  Google Scholar 

  31. Lavin MF. Ataxia-telangiectasia: from a rare disorder to a paradigm for cell signalling and cancer. Nat Rev Mol Cell Biol. 2008;9(10):759–69.

    Article  CAS  PubMed  Google Scholar 

  32. Lavin MF. Ataxia-telangiectasia: from a rare disorder to a paradigm for cell signalling and cancer. Nat Rev Mol Cell Biol. 2008;9(10):759–69.

    Article  CAS  PubMed  Google Scholar 

  33. Marzullo F, Simone G, Labriola A, Paradiso A, Rella G, Petroni S. Ki67 as histochemical marker of cellular proliferative activity in breast cancer. Pathologica. 1988;80(1067):279–85.

    CAS  PubMed  Google Scholar 

  34. Yang Y, Durando M, Smith-Roe SL, Sproul C, Greenwalt AM, Kaufmann W, et al. Cell cycle stage-specific roles of Rad18 in tolerance and repair of oxidative DNA damage. Nucleic Acids Res. 2013;41(4):2296–312.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Huang J, Huen MS, Kim H, Leung CC, Glover JN, Yu X, et al. RAD18 transmits DNA damage signalling to elicit homologous recombination repair. Nat Cell Biol. 2009;11(5):592–603.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Inagaki A, van Cappellen WA, van der Laan R, Houtsmuller AB, Hoeijmakers JH, Grootegoed JA, et al. Dynamic localization of human RAD18 during the cell cycle and a functional connection with DNA double-strand break repair. DNA Repair (Amst). 2009;8(2):190–201.

    Article  CAS  Google Scholar 

  37. Watanabe K, Iwabuchi K, Sun J, Tsuji Y, Tani T, Tokunaga K, et al. RAD18 promotes DNA double-strand break repair during G1 phase through chromatin retention of 53BP1. Nucleic Acids Res. 2009;37(7):2176–93.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Palle K, Vaziri C. Rad18 E3 ubiquitin ligase activity mediates Fanconi anemia pathway activation and cell survival following DNA Topoisomerase 1 inhibition. Cell Cycle. 2011;10(10):1625–38.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Sachdeva M, Mo YY. MicroRNA-145 suppresses cell invasion and metastasis by directly targeting mucin 1. Cancer Res. 2010;70(1):378–87.

    Article  CAS  PubMed  Google Scholar 

  40. Zhang J, Guo H, Zhang H, Wang H, Qian G, Fan X, et al. Putative tumor suppressor miR-145 inhibits colon cancer cell growth by targeting oncogene Friend leukemia virus integration 1 gene. Cancer. 2011;117(1):86–95.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors thank Dr. Wen-Jun Wang for her support and assistance with the preparation of the manuscript.

Conflicts of Interest

No potential conflicts of interest were disclosed.

Authors’ contributions

Wei-Dong Li conceived and designed the experiments. Ye Dong and Yan-Zhen Deng performed the experiments. Ye Dong and Yan-Zhen Deng analyzed the data. Ye Dong, Wen-Jun Wang, and Wei-Dong Li wrote the paper.

Grant support

This study was supported by grants from the National Natural Science Foundation of China (no. 81301939). The costs of publication of this article were defrayed in part by the payment of page charges.

Author information

Author notes

    Authors and Affiliations

    1. Department of the First Medicine, Cancer Hospital of Guangzhou Medical College, Guangzhou, 510095, China

      Rui-Lei Liu, Ye Dong, Yan-Zhen Deng & Wei-Dong Li

    2. State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical University, Guangzhou, 510120, China

      Wen-Jun Wang

    Authors
    1. Rui-Lei Liu
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Ye Dong
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Yan-Zhen Deng
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Wen-Jun Wang
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Wei-Dong Li
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Wei-Dong Li.

    Additional information

    Rui-Lei Liu and Ye Dong contributed equally to this work.

    Electronic supplementary material

    Below is the link to the electronic supplementary material.

    Supplementary Methods

    (DOC 33 kb)

    Table S1

    (XLS 17 kb)

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Liu, RL., Dong, Y., Deng, YZ. et al. Tumor suppressor miR-145 reverses drug resistance by directly targeting DNA damage-related gene RAD18 in colorectal cancer. Tumor Biol. 36, 5011–5019 (2015). https://doi.org/10.1007/s13277-015-3152-5

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s13277-015-3152-5

    Keywords

    Search

    Navigation