Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Roundabout homolog 1 inhibits proliferation via the YY1-ROBO1-CCNA2-CDK2 axis in human pancreatic cancer

Oncogene volume 40, pages 2772–2784 (2021)Cite this article

Subjects

Abstract

Pancreatic cancer (PC) is highly malignant and has a high mortality with a 5-year survival rate of less than 8%. As a member of the roundabout immunoglobulin superfamily of proteins, ROBO1 plays an important role in embryogenesis and organogenesis and also inhibits metastasis in PC. Our study was designed to explore whether ROBO1 has effects on the proliferation of PC and its specific mechanism. The expression of ROBO1 was higher in cancer tissues than in matched adjacent tissues by immunohistochemistry (IHC) and qRT-PCR. Low ROBO1 expression is associated with PC progression and poor prognosis. Overexpression of ROBO1 can inhibit the proliferation of PC cells in vitro, and the S phase fraction can also be induced. Further subcutaneous tumor formation in nude mice showed that ROBO1 overexpression can significantly inhibit tumor growth. YY1 was found to directly bind to the promoter region of ROBO1 to promote transcription by a luciferase reporter gene assay, a chromatin immunoprecipitation (ChIP) and an electrophoretic mobility shift assay (EMSA). Mechanistic studies showed that YY1 can inhibit the development of PC by directly regulating ROBO1 via the CCNA2/CDK2 axis. Taken together, our results suggest that ROBO1 may be involved in the development and progression of PC by regulating cell proliferation and shows that ROBO1 may be a novel and promising therapeutic target for PC.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: ROBO1 expression in PC.
Fig. 2: ROBO1 affects the proliferation of PC cells.
Fig. 3: Interaction between ROBO1 and YY1.
Fig. 4: YY1 binds to the promoter region of ROBO1 to regulate its expression.
Fig. 5: ROBO1 can partially restore the effects of YY1 on the proliferation of pancreatic cancer cells.
Fig. 6: ROBO1 inhibits pancreatic cancer growth in vivo by targeting the CCNA2/CDK2 axis.

Similar content being viewed by others

Data availability

All data generated or analyzed during this study are included in this published article.

References

  1. Pommier A, Anaparthy N, Memos N, Kelley ZL, Gouronnec A, Yan R, et al. Unresolved endoplasmic reticulum stress engenders immune-resistant, latent pancreatic cancer metastases. Science. 2018;360:eaao4908.

  2. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. CA Cancer J Clin. 2018;68:7–30.

    Article  Google Scholar 

  3. Chantrill LA, Nagrial AM, Watson C, Johns AL, Martyn-Smith M, Simpson S, et al. Precision medicine for advanced pancreas cancer: the individualized molecular pancreatic cancer therapy (IMPaCT) trial. Clin Cancer Res. 2015;21:2029–37.

    Article  CAS  Google Scholar 

  4. Kamisawa T, Wood LD, Itoi T, Takaori K. Pancreatic cancer. Lancet. 2016;388:73–85.

    Article  CAS  Google Scholar 

  5. Pinho AV, Van Bulck M, Chantrill L, Arshi M, Sklyarova T, Herrmann D, et al. ROBO2 is a stroma suppressor gene in the pancreas and acts via TGF-beta signalling. Nat Commun. 2018;9:5083.

    Article  Google Scholar 

  6. Kong R, Yi F, Wen P, Liu J, Chen X, Ren J, et al. Myo9b is a key player in SLIT/ROBO-mediated lung tumor suppression. J Clin Invest. 2015;125:4407–20.

    Article  Google Scholar 

  7. Xia Y, Wang L, Xu Z, Kong R, Wang F, Yin K, et al. Reduced USP33 expression in gastric cancer decreases inhibitory effects of Slit2-Robo1 signalling on cell migration and EMT. Cell Prolif. 2019;52:e12606.

    Article  Google Scholar 

  8. Parray A, Siddique HR, Kuriger JK, Mishra SK, Rhim JS, Nelson HH, et al. ROBO1, a tumor suppressor and critical molecular barrier for localized tumor cells to acquire invasive phenotype: study in African-American and Caucasian prostate cancer models. Int J Cancer. 2014;135:2493–506.

    Article  CAS  Google Scholar 

  9. Qin F, Zhang H, Ma L, Liu X, Dai K, Li W, et al. Low expression of Slit2 and Robo1 is associated with poor prognosis and brain-specific metastasis of breast cancer patients. Sci Rep. 2015;5:14430.

    Article  CAS  Google Scholar 

  10. Gohrig A, Detjen KM, Hilfenhaus G, Korner JL, Welzel M, Arsenic R, et al. Axon guidance factor SLIT2 inhibits neural invasion and metastasis in pancreatic cancer. Cancer Res. 2014;74:1529–40.

    Article  Google Scholar 

  11. Zhang JJ, Zhu Y, Zhang XF, Liu DF, Wang Y, Yang C, et al. Yin Yang-1 suppresses pancreatic ductal adenocarcinoma cell proliferation and tumor growth by regulating SOX2OT-SOX2 axis. Cancer Lett. 2017;408:144–54.

    Article  CAS  Google Scholar 

  12. Zhang JJ, Zhu Y, Xie KL, Peng YP, Tao JQ, Tang J, et al. Yin Yang-1 suppresses invasion and metastasis of pancreatic ductal adenocarcinoma by downregulating MMP10 in a MUC4/ErbB2/p38/MEF2C-dependent mechanism. Mol Cancer. 2014;13:130.

    Article  Google Scholar 

  13. Gopinathan L, Tan SL, Padmakumar VC, Coppola V, Tessarollo L, Kaldis P. Loss of Cdk2 and cyclin A2 impairs cell proliferation and tumorigenesis. Cancer Res. 2014;74:3870–9.

    Article  CAS  Google Scholar 

  14. Liu J, Hou W, Guan T, Tang L, Zhu X, Li Y, et al. Slit2/Robo1 signaling is involved in angiogenesis of glomerular endothelial cells exposed to a diabetic-like environment. Angiogenesis. 2018;21:237–49.

    Article  CAS  Google Scholar 

  15. Leyva-Diaz E, del Toro D, Menal MJ, Cambray S, Susin R, Tessier-Lavigne M, et al. FLRT3 is a Robo1-interacting protein that determines Netrin-1 attraction in developing axons. Curr Biol. 2014;24:494–508.

    Article  CAS  Google Scholar 

  16. Tang W, Zhou W, Xiang L, Wu X, Zhang P, Wang J, et al. The p300/YY1/miR-500a-5p/HDAC2 signalling axis regulates cell proliferation in human colorectal cancer. Nat Commun. 2019;10:663.

    Article  CAS  Google Scholar 

  17. Patten DK, Corleone G, Gyorffy B, Perone Y, Slaven N, Barozzi I, et al. Enhancer mapping uncovers phenotypic heterogeneity and evolution in patients with luminal breast cancer. Nat Med. 2018;24:1469–80.

    Article  CAS  Google Scholar 

  18. Gao D, Wang L, Zhang H, Yan X, Yang J, Zhou R, et al. Spleen tyrosine kinase SYK(L) interacts with YY1 and coordinately suppresses SNAI2 transcription in lung cancer cells. FEBS J. 2018;285:4229–45.

    Article  CAS  Google Scholar 

  19. Liu D, Zhang J, Wu Y, Shi G, Yuan H, Lu Z, et al. YY1 suppresses proliferation and migration of pancreatic ductal adenocarcinoma by regulating the CDKN3/MdM2/P53/P21 signaling pathway. Int J Cancer. 2018;142:1392–404.

    Article  CAS  Google Scholar 

  20. Dachineni R, Ai G, Kumar DR, Sadhu SS, Tummala H, Bhat GJ. Cyclin A2 and CDK2 as novel targets of aspirin and salicylic acid: a potential role in cancer prevention. Mol Cancer Res. 2016;14:241–52.

    Article  CAS  Google Scholar 

  21. Zhang QH, Yuen WS, Adhikari D, Flegg JA, FitzHarris G, Conti M, et al. Cyclin A2 modulates kinetochore-microtubule attachment in meiosis II. J Cell Biol. 2017;216:3133–43.

    Article  CAS  Google Scholar 

  22. Honda A, Valogne Y, Bou Nader M, Brechot C, Faivre J. An intron-retaining splice variant of human cyclin A2, expressed in adult differentiated tissues, induces a G1/S cell cycle arrest in vitro. PLoS One. 2012;7:e39249.

    Article  CAS  Google Scholar 

  23. Wang B, Li D, Kovalchuk A, Litvinov D, Kovalchuk O. Ionizing radiation-inducible miR-27b suppresses leukemia proliferation via targeting cyclin A2. Int J Radiat Oncol Biol Phys. 2014;90:53–62.

    Article  CAS  Google Scholar 

  24. Peng X, Pan K, Zhao W, Zhang J, Yuan S, Wen X, et al. NPTX1 inhibits colon cancer cell proliferation through down-regulating cyclin A2 and CDK2 expression. Cell Biol Int. 2018;42:589–97.

    Article  CAS  Google Scholar 

  25. Moore NL, Edwards DP, Weigel NL. Cyclin A2 and its associated kinase activity are required for optimal induction of progesterone receptor target genes in breast cancer cells. J Steroid Biochem Mol Biol. 2014;144(Pt B):471–82.

    Article  CAS  Google Scholar 

Download references

Funding

This study was supported by the National Natural Science Foundation of China (Nos. 81871980, 81572337, 81672449); the National Science Foundation for Distinguished Young Scholars of China (No. 81902455); the Jiangsu Key Medical Discipline (General Surgery; ZDXKA2016005); the Innovation Capability Development Project of Jiangsu Province (No. BM2015004); the Priority Academic Program AQ3 Development of Jiangsu Higher Education Institutions (PAPD, JX10231801) and the Project of Invigorating Health Care through Science, Technology and Education, Jiangsu Provincial Medical Outstanding Talent (to Yi Miao, JCRCA2016009).

Author information

Author notes

  1. These authors contributed equally: Qun Chen, Peng Shen, Wan-Li Ge, Tao-Yue Yang

Authors and Affiliations

  1. Pancreas Center, the First Affiliated Hospital of Nanjing Medical University, Nanjing, China

    Qun Chen, Peng Shen, Wan-Li Ge, Tao-Yue Yang, Ling-Dong Meng, Xu-Min Huang, Yi-Han Zhang, Shou-Ji Cao, Yi Miao, Kui-Rong Jiang & Jing-Jing Zhang

  2. Pancreas Institute, Nanjing Medical University, Nanjing, China

    Qun Chen, Peng Shen, Wan-Li Ge, Tao-Yue Yang, Ling-Dong Meng, Xu-Min Huang, Yi-Han Zhang, Shou-Ji Cao, Yi Miao, Kui-Rong Jiang & Jing-Jing Zhang

  3. Nanjing University of Chinese Medicine, Nanjing, China

    Wu-Jun Wang

Authors
  1. Qun Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Peng Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Wan-Li Ge
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tao-Yue Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wu-Jun Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ling-Dong Meng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Xu-Min Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Yi-Han Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shou-Ji Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Yi Miao
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Kui-Rong Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Jing-Jing Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

QC, PS, WLG and TYY carried out the studies, participated in the experimental design, statistical analysis and drafted the manuscript. LDM and WJW participated in the luciferase reporter assays and ChIP assays. XMH and YHZ participated in the qRT-PCR and Western blotting. SJC participated in the sample collection, patient follow-up and statistical analysis. QC and WJW carried out the in vivo studies and participated in the statistical analysis. KRJ, JJZ and YM critically revised the manuscript for important intellectual content. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Kui-Rong Jiang or Jing-Jing Zhang.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Ethical approval

This study was approved by the Ethics Committee of the First Affiliated Hospital with Nanjing Medical University.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, Q., Shen, P., Ge, WL. et al. Roundabout homolog 1 inhibits proliferation via the YY1-ROBO1-CCNA2-CDK2 axis in human pancreatic cancer. Oncogene 40, 2772–2784 (2021). https://doi.org/10.1038/s41388-021-01741-5

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41388-021-01741-5

This article is cited by

Search

Advanced search

Quick links