The CTBP2-PCIF1 complex regulates m6Am modification of mRNA in head and neck squamous cell carcinoma
Kang Li, … , Qianming Chen, Demeng Chen
Kang Li, … , Qianming Chen, Demeng Chen
Published August 29, 2023
Citation Information: J Clin Invest. 2023;133(20):e170173. https://doi.org/10.1172/JCI170173.
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Research Article Cell biology Oncology

The CTBP2-PCIF1 complex regulates m6Am modification of mRNA in head and neck squamous cell carcinoma

Abstract

PCIF1 can mediate the methylation of N6,2′-O-dimethyladenosine (m6Am) in mRNA. Yet, the detailed interplay between PCIF1 and the potential cofactors and its pathological significance remain elusive. Here, we demonstrated that PCIF1-mediated cap mRNA m6Am modification promoted head and neck squamous cell carcinoma progression both in vitro and in vivo. CTBP2 was identified as a cofactor of PCIF1 to catalyze m6Am deposition on mRNA. CLIP-Seq data demonstrated that CTBP2 bound to similar mRNAs as compared with PCIF1. We then used the m6Am-Seq method to profile the mRNA m6Am site at single-base resolution and found that mRNA of TET2, a well-known tumor suppressor, was a major target substrate of the PCIF1-CTBP2 complex. Mechanistically, knockout of CTBP2 reduced PCIF1 occupancy on TET2 mRNA, and the PCIF1-CTBP2 complex negatively regulated the translation of TET2 mRNA. Collectively, our study demonstrates the oncogenic function of the epitranscriptome regulator PCIF1-CTBP2 complex, highlighting the importance of the m6Am modification in tumor progression.

Authors

Kang Li, Jie Chen, Caihua Zhang, Maosheng Cheng, Shuang Chen, Wei Song, Chunlong Yang, Rongsong Ling, Zhi Chen, Xiaochen Wang, Gan Xiong, Jieyi Ma, Yan Zhu, Quan Yuan, Qi Liu, Liang Peng, Qianming Chen, Demeng Chen

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Figure 2

Knockout of PCIF1 suppresses the HNSCC malignant phenotype.

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Knockout of PCIF1 suppresses the HNSCC malignant phenotype. (A) Western ...
(A) Western blotting analyses of the PCIF1 expression in the cell lines. (B) Western blotting analyses detecting the PCIF1 expression in SCC9 (left) and SCC25 (right) control cells and PCIF1-KO cells. (C) Cell Counting Kit-8 (CCK8) assay of cell viability in control and PCIF1-KO cells (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 by 1-way ANOVA, Dunnett’s test. (D) Colony formation assay detecting the colony ability of control and PCIF1-KO cells (n = 3). ***P < 0.001 by 1-way ANOVA with Tukey’s multiple-comparison test. (E and F) The cell migration (E) and invasion (F) ability of control and PCIF1-KO cells was determined by Transwell assay (n = 3). **P < 0.01, ***P < 0.001 by 1-way ANOVA with Tukey’s multiple-comparison test. Scale bar: 100 μm. (G) Flow cytometry assay for cell apoptosis in control and PCIF1-KO cells. Bottom: Representative images. Top: Quantification data (n = 3). ***P < 0.001 by 1-way ANOVA with Tukey’s multiple-comparison test. (H) Cell cycle progression was detected by flow cytometric analyses in control and PCIF1-KO cells (n = 3). **P < 0.01, ***P < 0.001 by 1-way ANOVA with Tukey’s multiple-comparison test.