Open Accessm 6 A‐mediated regulation of PBX1‐GCH1 axis promotes gastric cancer proliferation and metastasis by elevating tetrahydrobiopterin levels
Author(s) -
Liu Yinan,
Zhai Ertao,
Chen Junting,
Qian Yan,
Zhao Risheng,
Ma Yan,
Liu Jianqiu,
Huang Zhixin,
Cai Shirong,
Chen Jianhui
Publication year - 2022
Publication title -
cancer communications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.119
H-Index - 53
ISSN - 2523-3548
DOI - 10.1002/cac2.12281
Subject(s) - gene knockdown , chromatin immunoprecipitation , tetrahydrobiopterin , cancer research , chemistry , biology , nitric oxide synthase , cell culture , gene expression , promoter , biochemistry , enzyme , genetics , gene
Background Methyltransferase 3 (METTL3)‐mediated N6‐methyladenosine (m 6 A) RNA modification has been demonstrated to be a potential factor in promoting gastric cancer (GC). METTL3 regulates a series of signaling pathways by modifying various mRNAs. This study aimed to identify novel METTL3‐mediated signaling pathways and explored possible targets for use in the clinical setting of gastric cancer. Methods To investigate the proliferation and metastatic capacity of GC cell lines with METTL3 knockdown, a xenograft, lung metastasis, and popliteal lymph node metastasis model was used. The m 6 A‐modified RNA immunoprecipitation (Me‐RIP) sequence was utilized to explore the target mRNAs of METTL3. Cell counting kit 8 and transwell assays were performed to investigate the promoting function of pre‐B cell leukemia homeobox 1 (PBX1) and GTP cyclohydrolase 1 (GCH1). Western blotting and chromatin immunoprecipitation were employed to confirm the involvement of the METTL3‐PBX1‐GCH1 axis. ELISA and liquid chromatography‐mass spectrometry were used to explore the biological function of tetrahydrobiopterin (BH 4 ). Results Knockdown of METTL3 suppressed xenograft tumor growth and lung/lymph node metastasis in vivo. Mechanistically, we found that METTL3 combined with and stabilized PBX1 mRNAs. Chromatin immunoprecipitation (ChIP) and further experiments suggested that PBX1 acted as a transcription factor inducing GCH1 expression. Moreover, the METTL3‐PBX1‐GCH1 axis increased BH 4 levels in GC cells, thereby promoting tumor progression. Conclusions This study suggested that METTL3 enzymes promote tumor growth and lung/lymph node metastasis via METTL3‐PBX1‐GCH1 axis increasing BH4 levels in GC.