Abstract
Cancer stem cells (CSCs) encompass a subset of highly aggressive tumor cells that are involved in tumor initiation and progression. This study investigates the function of regulator of calcineurin 2 (RCAN2) in the stem cell property in colorectal cancer (CRC). By analyzing four GEO datasets, we obtained RCAN2 as a stemness-related gene in CRC. RCAN2 was poorly expressed in CRC tissues and cells, especially in CSCs. RCAN2 restoration reduced calcineurin activity and promoted phosphorylation and degradation of nuclear factor of activated T cells 1 (NFATC1) protein, leading to reduced stemness of CSCs. JunD proto-oncogene (JUND), whose protein level was increased in CRC samples and CRC stem cells, bound to RCAN2 and suppressed its transcription. The abundant ubiquitin specific peptidase 7 (USP7) in CSCs enhanced JUND protein stability through deubiquitination modification. Lentivirus-mediated knockdown of USP7 or JUND also blocked the calcineurin-NFATC1 signaling and reduced the protein levels of stemness-related proteins. Moreover, the USP7 knockdown weakened the colony/sphere formation ability as well as the tumorigenicity of CSCs, and it reduced the CSC content in xenograft tumors. However, further restoration of JUND rescued the stemness of the CSCs. Overall, this study demonstrates that USP7-mediated JUND suppresses RCAN2 transcription and activates NFATC1 to enhance stem cell property in CRC.
Graphical abstract
1. RCAN2 is poorly expressed in CRC tissues and cells and especially in CSCs.
2. RCAN2 reduces stemness of CSCs by blocking calcineurin-NFATC1 signal transduction.
3. JUND binds to RCAN2 promoter to suppresses RCAN2 transcription.
4. USP7 enhances JUND protein stability via deubiquitination modification.
5. Downregulation of USP7 or JUND restores RCAN2 level and suppresses stemness of CSCs.
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All data generated or analyzed during this study are included in this article. Additional raw data may be available from the corresponding author for reasonable reasons.
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Funding
This work was supported by Pudong New Area Science and Technology Development Fund for People’s Livelihood Research Projects of Public Institutions (Medical and Health Projects) (No. PKJ2021-Y24).
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Y.L.C. wrote the original draft, participated in the design of the study, and performed the experiments. L.L.C. performed the experiments, data collection, and interpretation. J.T. and G.Y.C. performed the experiments and analyzed the data. J.R.J. and M.X. provided resources, administration supports, and data amendment. All authors read and approved the final manuscript.
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All patients signed the informed consent form. The present study was approved by the Ethics Committee of the Pudong New Area People’s Hospital (No. PAPH20210122).
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Chang, Y., Chen, L., Tang, J. et al. USP7-mediated JUND suppresses RCAN2 transcription and elevates NFATC1 to enhance stem cell property in colorectal cancer. Cell Biol Toxicol 39, 3121–3140 (2023). https://doi.org/10.1007/s10565-023-09822-9
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DOI: https://doi.org/10.1007/s10565-023-09822-9