m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47

Abstract Background Hepatocellular carcinoma (HCC) is one of most common malignancies with poor prognosis, largely due to its high propensity for metastasis and recurrence. As the most common internal RNA modification, N6-methyladenosine (m6A) plays critical and diverse roles in HCC progression. How...

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Main Authors: Huamei Wei, Changhong Pu, Min Zeng, Rongzhou Lu, Yunyu Wei, Yanyan Huang, Zheng Huang, Lizheng Huang, Zuoming Xu, Jianchu Wang, Rihai Ma, Jian Pu
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Cancer Cell International
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Online Access:https://doi.org/10.1186/s12935-025-03857-0
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author Huamei Wei
Changhong Pu
Min Zeng
Rongzhou Lu
Yunyu Wei
Yanyan Huang
Zheng Huang
Lizheng Huang
Zuoming Xu
Jianchu Wang
Rihai Ma
Jian Pu
author_facet Huamei Wei
Changhong Pu
Min Zeng
Rongzhou Lu
Yunyu Wei
Yanyan Huang
Zheng Huang
Lizheng Huang
Zuoming Xu
Jianchu Wang
Rihai Ma
Jian Pu
author_sort Huamei Wei
collection DOAJ
description Abstract Background Hepatocellular carcinoma (HCC) is one of most common malignancies with poor prognosis, largely due to its high propensity for metastasis and recurrence. As the most common internal RNA modification, N6-methyladenosine (m6A) plays critical and diverse roles in HCC progression. However, the mechanisms by which m6A regulates HCC metastasis remain incompletely understood. Here, we aimed to identify key m6A modification events during HCC metastasis. Methods The expression of CTC-297N7.9. was measured by qPCR. m6A modification level of CTC-297N7.9 was measured by methylated RNA immunoprecipitation (MeRIP) and single-base elongation- and ligation-based qPCR amplification method (SELECT). The roles of m6A-modified CTC-297N7.9 in HCC were investigated by in vitro cell viability, proliferation, migration and phagocytosis assays, and in vivo liver metastasis and lung metastasis assays. The underlying mechanisms of m6A-modified CTC-297N7.9 were dissected by chromatin isolation by RNA purification (ChIRP), assay for transposase accessible chromatin (ATAC) and cleavage under target & tagmentation (CUT&Tag) assays. Results The m6A modification level of CTC-297N7.9 is decreased in HCC tissues and correlated with microvascular invasion and poor prognosis. CTC-297N7.9 suppresses HCC metastasis in an m6A-dependent manner. m6A-modified CTC-297N7.9 attenuates tumor-associated macrophages (TAMs) infiltration and M2 polarization through downregulating CCL2 expression and secretion. Additionally, m6A-modified CTC-297N7.9 promotes phagocytosis of HCC cells by macrophages through downregulating the phagocytosis checkpoint CD47. Mechanistic investigations revealed that m6A-modified CTC-297N7.9 binds the m6A reader YTHDC1, which further binds and recruits the histone H3K9me3 methyltransferase SETDB1 and H3K27me3 methyltransferase EZH2 to the promoters of CCL2 and CD47, leading to the upregulation of H3K9me3 and H3K27me3 modifications at CCL2 and CD47 promoters, and transcriptional silencing of CCL2 and CD47. Conclusions Our study demonstrates that m6A-modified CTC-297N7.9 acts as a metastasis suppressor in HCC, and highlights its potential as a prognostic biomarker and therapeutic target for HCC.
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publishDate 2025-07-01
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series Cancer Cell International
spelling doaj-art-dc8f68e3f0da45fc8a0a1a72cb05d1da2025-08-20T03:06:06ZengBMCCancer Cell International1475-28672025-07-0125111910.1186/s12935-025-03857-0m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47Huamei Wei0Changhong Pu1Min Zeng2Rongzhou Lu3Yunyu Wei4Yanyan Huang5Zheng Huang6Lizheng Huang7Zuoming Xu8Jianchu Wang9Rihai Ma10Jian Pu11Department of Pathology, Affiliated Hospital of Youjiang Medical University for NationalitiesDepartment of Radiology, Affiliated Hospital of Youjiang Medical University for NationalitiesGraduate College of Youjiang Medical University for NationalitiesGraduate College of Youjiang Medical University for NationalitiesGraduate College of Youjiang Medical University for NationalitiesGraduate College of Youjiang Medical University for NationalitiesGraduate College of Youjiang Medical University for NationalitiesGraduate College of Youjiang Medical University for NationalitiesDepartment of Hepatobiliary Surgery, Affiliated Hospital of Youjiang Medical University for NationalitiesGuangxi Clinical Medical Research Center of Hepatobiliary DiseaseDepartment of Hepatobiliary Surgery, Affiliated Hospital of Youjiang Medical University for NationalitiesGuangxi Clinical Medical Research Center of Hepatobiliary DiseaseAbstract Background Hepatocellular carcinoma (HCC) is one of most common malignancies with poor prognosis, largely due to its high propensity for metastasis and recurrence. As the most common internal RNA modification, N6-methyladenosine (m6A) plays critical and diverse roles in HCC progression. However, the mechanisms by which m6A regulates HCC metastasis remain incompletely understood. Here, we aimed to identify key m6A modification events during HCC metastasis. Methods The expression of CTC-297N7.9. was measured by qPCR. m6A modification level of CTC-297N7.9 was measured by methylated RNA immunoprecipitation (MeRIP) and single-base elongation- and ligation-based qPCR amplification method (SELECT). The roles of m6A-modified CTC-297N7.9 in HCC were investigated by in vitro cell viability, proliferation, migration and phagocytosis assays, and in vivo liver metastasis and lung metastasis assays. The underlying mechanisms of m6A-modified CTC-297N7.9 were dissected by chromatin isolation by RNA purification (ChIRP), assay for transposase accessible chromatin (ATAC) and cleavage under target & tagmentation (CUT&Tag) assays. Results The m6A modification level of CTC-297N7.9 is decreased in HCC tissues and correlated with microvascular invasion and poor prognosis. CTC-297N7.9 suppresses HCC metastasis in an m6A-dependent manner. m6A-modified CTC-297N7.9 attenuates tumor-associated macrophages (TAMs) infiltration and M2 polarization through downregulating CCL2 expression and secretion. Additionally, m6A-modified CTC-297N7.9 promotes phagocytosis of HCC cells by macrophages through downregulating the phagocytosis checkpoint CD47. Mechanistic investigations revealed that m6A-modified CTC-297N7.9 binds the m6A reader YTHDC1, which further binds and recruits the histone H3K9me3 methyltransferase SETDB1 and H3K27me3 methyltransferase EZH2 to the promoters of CCL2 and CD47, leading to the upregulation of H3K9me3 and H3K27me3 modifications at CCL2 and CD47 promoters, and transcriptional silencing of CCL2 and CD47. Conclusions Our study demonstrates that m6A-modified CTC-297N7.9 acts as a metastasis suppressor in HCC, and highlights its potential as a prognostic biomarker and therapeutic target for HCC.https://doi.org/10.1186/s12935-025-03857-0N6-methyladenosineHepatocellular carcinomaMetastasisMacrophagePhagocytosisEpigenetic modulation
spellingShingle Huamei Wei
Changhong Pu
Min Zeng
Rongzhou Lu
Yunyu Wei
Yanyan Huang
Zheng Huang
Lizheng Huang
Zuoming Xu
Jianchu Wang
Rihai Ma
Jian Pu
m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47
Cancer Cell International
N6-methyladenosine
Hepatocellular carcinoma
Metastasis
Macrophage
Phagocytosis
Epigenetic modulation
title m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47
title_full m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47
title_fullStr m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47
title_full_unstemmed m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47
title_short m6A-modified CTC-297N7.9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating CCL2 and CD47
title_sort m6a modified ctc 297n7 9 inhibits hepatocellular carcinoma metastasis via epigenetically downregulating ccl2 and cd47
topic N6-methyladenosine
Hepatocellular carcinoma
Metastasis
Macrophage
Phagocytosis
Epigenetic modulation
url https://doi.org/10.1186/s12935-025-03857-0
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