A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growth

Abstract Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer that lacks reliable targets for diagnosis and therapy. Non-coding RNA (ncRNA)-encoded products hold promise for addressing this unmet need. By analyzing the reported ribosomal RNA sequencing data, combined w...

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Main Authors: Huichun Liang, Fubing Li, Huan Fang, Wenlong Ren, Zhongmei Zhou, Jiecheng Wang, Jialing Liu, Yongjia Tang, Xue Liu, Yingying Wu, Jing Peng, Chuanyu Yang, Jiayi Chen, Yuting Fei, Yujie Shi, Dewei Jiang, Nu Zhang, Ceshi Chen
Format: Article
Language:English
Published: Nature Publishing Group 2025-07-01
Series:Signal Transduction and Targeted Therapy
Online Access:https://doi.org/10.1038/s41392-025-02298-5
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author Huichun Liang
Fubing Li
Huan Fang
Wenlong Ren
Zhongmei Zhou
Jiecheng Wang
Jialing Liu
Yongjia Tang
Xue Liu
Yingying Wu
Jing Peng
Chuanyu Yang
Jiayi Chen
Yuting Fei
Yujie Shi
Dewei Jiang
Nu Zhang
Ceshi Chen
author_facet Huichun Liang
Fubing Li
Huan Fang
Wenlong Ren
Zhongmei Zhou
Jiecheng Wang
Jialing Liu
Yongjia Tang
Xue Liu
Yingying Wu
Jing Peng
Chuanyu Yang
Jiayi Chen
Yuting Fei
Yujie Shi
Dewei Jiang
Nu Zhang
Ceshi Chen
author_sort Huichun Liang
collection DOAJ
description Abstract Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer that lacks reliable targets for diagnosis and therapy. Non-coding RNA (ncRNA)-encoded products hold promise for addressing this unmet need. By analyzing the reported ribosomal RNA sequencing data, combined with the TCGA, ORFfinder, SmProt databases, we identified CDKN2B-AS1, a TNBC-upregulated lncRNA encoding a 66-amino-acid peptide via CUG-initiated translation. CRISPR-Cas9 gene editing and mass spectrometry confirmed endogenous expression of this peptide, designated 66CTG, in TNBC cells. Functionally independently of its host RNA, 66CTG promoted the proliferation of TNBC cells and the tumor growth of TNBC xenograft by stabilizing c-Myc protein and enhancing Cyclin D1 transcription. Immunohistochemistry of 89 clinical TNBC paraffin samples revealed positive correlations among 66CTG, c-Myc, and Cyclin D1 expression levels. Mechanistically, co-immunoprecipitation and ubiquitination assays revealed that 66CTG stabilized c-Myc by competitively interacting with FBW7α, an E3 ligase responsible for recognizing 66CTG CPDS56/S60 motif which phosphorylated by GSK-3β during the late G1 phase. In conclusion, our findings suggest 66CTG has potential to be developed as a target for TNBC diagnosis and therapy. Furthermore, it unveils a regulatory axis wherein 66CTG stabilizes c-Myc by interacting with FBW7α, offering a new mechanistic explanation for c-Myc overexpression in TNBC. Patients co-overexpressing 66CTG, c-Myc, and Cyclin D1 may benefit from therapies targeting this axis.
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spelling doaj-art-e1887e3c6245438eb7b46f186cac98ca2025-08-20T03:06:09ZengNature Publishing GroupSignal Transduction and Targeted Therapy2059-36352025-07-0110111610.1038/s41392-025-02298-5A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growthHuichun Liang0Fubing Li1Huan Fang2Wenlong Ren3Zhongmei Zhou4Jiecheng Wang5Jialing Liu6Yongjia Tang7Xue Liu8Yingying Wu9Jing Peng10Chuanyu Yang11Jiayi Chen12Yuting Fei13Yujie Shi14Dewei Jiang15Nu Zhang16Ceshi Chen17Key Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Chinese Academy of SciencesYunnan Key Laboratory of Breast Cancer Precision Medicine, Academy of Biomedical Engineering, Kunming Medical UniversityKey Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Chinese Academy of SciencesKey Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Chinese Academy of SciencesThe School of Continuing Education, Kunming Medical UniversityYunnan Key Laboratory of Breast Cancer Precision Medicine, The Third Affiliated Hospital, Kunming Medical UniversityYunnan Key Laboratory of Breast Cancer Precision Medicine, The Third Affiliated Hospital, Kunming Medical UniversityHangzhou Institute of Medicine, Chinese Academy of SciencesCenter of Medical Laboratory, General Hospital of Ningxia Medical UniversityDepartment of Pathology, The First Affiliated Hospital, Kunming Medical UniversityYunnan Key Laboratory of Breast Cancer Precision Medicine, Academy of Biomedical Engineering, Kunming Medical UniversityKey Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Chinese Academy of SciencesYunnan Key Laboratory of Breast Cancer Precision Medicine, The Third Affiliated Hospital, Kunming Medical UniversityYunnan Key Laboratory of Breast Cancer Precision Medicine, The Third Affiliated Hospital, Kunming Medical UniversityDepartment of Pathology, Henan Provincial People’s Hospital, Zhengzhou UniversityKey Laboratory of Animal Models and Human Disease Mechanisms of the Chinese Academy of Sciences and Yunnan Province, Kunming Institute of Zoology, Chinese Academy of SciencesDepartment of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangdong Provincial Key Laboratory of Brain Function and Disease, Guangdong Translational Medicine Innovation PlatformYunnan Key Laboratory of Breast Cancer Precision Medicine, Academy of Biomedical Engineering, Kunming Medical UniversityAbstract Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer that lacks reliable targets for diagnosis and therapy. Non-coding RNA (ncRNA)-encoded products hold promise for addressing this unmet need. By analyzing the reported ribosomal RNA sequencing data, combined with the TCGA, ORFfinder, SmProt databases, we identified CDKN2B-AS1, a TNBC-upregulated lncRNA encoding a 66-amino-acid peptide via CUG-initiated translation. CRISPR-Cas9 gene editing and mass spectrometry confirmed endogenous expression of this peptide, designated 66CTG, in TNBC cells. Functionally independently of its host RNA, 66CTG promoted the proliferation of TNBC cells and the tumor growth of TNBC xenograft by stabilizing c-Myc protein and enhancing Cyclin D1 transcription. Immunohistochemistry of 89 clinical TNBC paraffin samples revealed positive correlations among 66CTG, c-Myc, and Cyclin D1 expression levels. Mechanistically, co-immunoprecipitation and ubiquitination assays revealed that 66CTG stabilized c-Myc by competitively interacting with FBW7α, an E3 ligase responsible for recognizing 66CTG CPDS56/S60 motif which phosphorylated by GSK-3β during the late G1 phase. In conclusion, our findings suggest 66CTG has potential to be developed as a target for TNBC diagnosis and therapy. Furthermore, it unveils a regulatory axis wherein 66CTG stabilizes c-Myc by interacting with FBW7α, offering a new mechanistic explanation for c-Myc overexpression in TNBC. Patients co-overexpressing 66CTG, c-Myc, and Cyclin D1 may benefit from therapies targeting this axis.https://doi.org/10.1038/s41392-025-02298-5
spellingShingle Huichun Liang
Fubing Li
Huan Fang
Wenlong Ren
Zhongmei Zhou
Jiecheng Wang
Jialing Liu
Yongjia Tang
Xue Liu
Yingying Wu
Jing Peng
Chuanyu Yang
Jiayi Chen
Yuting Fei
Yujie Shi
Dewei Jiang
Nu Zhang
Ceshi Chen
A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growth
Signal Transduction and Targeted Therapy
title A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growth
title_full A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growth
title_fullStr A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growth
title_full_unstemmed A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growth
title_short A novel peptide 66CTG stabilizes Myc proto-oncogene protein to promote triple-negative breast cancer growth
title_sort novel peptide 66ctg stabilizes myc proto oncogene protein to promote triple negative breast cancer growth
url https://doi.org/10.1038/s41392-025-02298-5
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