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CircN4BP2L2 promotes colorectal cancer growth and metastasis through regulation of the miR-340-5p/CXCR4 axis

Abstract

Colorectal cancer (CRC) is the third leading cause of cancer-related death worldwide. Dysregulation of circular RNAs (circRNAs) appears to be a critical factor in CRC progression. However, mechanistic studies delineating the role of circRNAs in CRC remain limited. In this study, qRT-PCR and western blot assays were used to measure the expression of genes and proteins. Migration, invasion, proliferation, and apoptosis were examined by wound-healing, transwell, CCK-8, colony formation, and flow cytometry assays, respectively. Molecular interactions were validated by a dual-luciferase report system. A xenograft animal model was established to examine in vivo tumor growth and lung metastasis. Our data indicated that circN4BP2L2 expression was increased in CRC tissues and cell lines. Notably, inhibition of circN4BP2L2 effectively inhibited proliferation, migration, and invasion of LoVo cells, and inhibited tumor growth and metastasis in vivo, whereas the forced expression of circN4BP2L2 facilitated the proliferation, migration, and invasion of HT-29 cells. Mechanistic studies revealed that circN4BP2L2 acted as a molecular sponge of miR-340-5p to competitively promote CXCR4 expression. Furthermore, inhibition of miR-340-5p reversed the anti-cancer effects of circN4BP2L2 or CXCR4 silencing. Our data indicated an oncogenic role of circN4BP2L2 in CRC via regulation of the miR-340-5p/CXCR4 axis, which may be a promising biomarker and target for CRC treatment.

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Fig. 1: Upregulation of circN4BP2L2 in CRC tissues and cell lines.
Fig. 2: Silencing of circN4BP2L2 impairs cell proliferation, migration and invasion, and induces apoptosis of CRC cells in vitro.
Fig. 3: Forced expression of circN4BP2L2 promotes cell proliferation, migration, and invasion, but inhibits apoptosis in CRC cells.
Fig. 4: Silencing of circN4BP2L suppressed tumor growth and CRC lung metastasis in vivo.
Fig. 5: circN4BP2L2 enhances CXCR4 expression by acting as a molecular sponge of miR-340-5p.
Fig. 6: Knockdown of circN4BP2L2 suppresses CRC cell growth and metastasis through regulating miR-340-5p/CXCR4 axis.
Fig. 7: circN4BP2L2 promotes CRC cell growth and metastasis by regulating the miR-340-5p/CXCR4 axis.
Fig. 8: Schematic diagram of research mechanism.

Data availability

The datasets generated and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Authors

Contributions

KY and YW performed the experiments and wrote the manuscript. FZ collected and analyzed the data. ZZ designed and supervised the study. BL contributed the methodology. DF and ZZ analyzed the data and edited the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Zhi-Jun Zeng.

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The study has been approved by the Hospital’s Ethics Committee. All patients were informed of the study and signed the written consent.

Funding statement

This work was supported by the Natural Science Foundation of Hunan Province (No. 2019JJ40480).

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The authors declare no competing interests.

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Yang, KD., Wang, Y., Zhang, F. et al. CircN4BP2L2 promotes colorectal cancer growth and metastasis through regulation of the miR-340-5p/CXCR4 axis. Lab Invest (2021). https://doi.org/10.1038/s41374-021-00632-3

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