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

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.

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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.

Data availability

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

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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).

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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.

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The authors declare no competing interests.

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

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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

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