Kun Qian1, Wei Xu2, Xiaoyao Xia1, Jinhuo Ding3,4. 1. Department of General Surgery, Shangrao People's Hospital, 334000, Shangrao, China. 2. Department of Gastrointestinal Surgery, Colorectal Tumor Minimally Invasive Center, The Second Affiliated Hospital of Nanchang University, 330000, Nanchang, China. 3. Department of General Surgery, Shangrao People's Hospital, 334000, Shangrao, China. djhsr88@163.com. 4. Department of General Surgery, Shangrao People's Hospital, No. 86 Shuyuan Road, 334000, Shangrao City, Jiangxi Province Shangrao, China. djhsr88@163.com.
Abstract
BACKGROUND: Methyltransferase-like 3 (METTL3) is an RNA N6-methyladenosine (m6A) methyltransferase, which plays a critical role in micorRNA (miRNAs) processing and maturation, but it is still unclear whether METTL3 regulated miRNAs participates in the regulation of cancer aggressiveness in gastrointestinal stromal tumors (GISTs). OBJECTIVES: This study was designed to investigate this issue, and uncover the potential underlying mechanisms. METHODS: the expression of METTL3 in GISTs tissues and cell lines were determined by RT-qPCR and Western blot. Cell proliferation and migration were assessed by colony formation, CCK-8 and Transwell. The mRNA expression of all proteins was detected by RT-qPCR, and tumor xenograft study was applied to confirm the effect of METTL3 on GISTs development in vivo. RESULTS: In our study, we showed that METTL3 was significantly upregulated in GISTs tissues and cell lines. Functional experiments demonstrated that overexpression of METTL3 promoted GISTs cell malignant biological behavior and tumor growth in vitro and in vivo, and conversely, silencing of METTL3 had opposite effects and suppressed GISTs progression. Further mechanistical experiments verified that METTL3 promoted the maturation of miR-25-3p in an m6A-dependent manner. Similar to METTL3, miR-25-3p was also validated as an oncogene to promote cancer development in GISTs. Finally, our rescuing experiments hinted that silencing of miR-25-3p abrogated the tumor-initiating effects of METTL3 overexpression on GISTs. CONCLUSION: Collectively, those results indicated that METTL3 played an oncogenic role in GISTs through positively modulating the miR-25-3p in an m6A-dependent manner, and we firstly discussed how the METTL3/m6A/miR-25-3p axis affected GISTs development.
BACKGROUND: Methyltransferase-like 3 (METTL3) is an RNA N6-methyladenosine (m6A) methyltransferase, which plays a critical role in micorRNA (miRNAs) processing and maturation, but it is still unclear whether METTL3 regulated miRNAs participates in the regulation of cancer aggressiveness in gastrointestinal stromal tumors (GISTs). OBJECTIVES: This study was designed to investigate this issue, and uncover the potential underlying mechanisms. METHODS: the expression of METTL3 in GISTs tissues and cell lines were determined by RT-qPCR and Western blot. Cell proliferation and migration were assessed by colony formation, CCK-8 and Transwell. The mRNA expression of all proteins was detected by RT-qPCR, and tumor xenograft study was applied to confirm the effect of METTL3 on GISTs development in vivo. RESULTS: In our study, we showed that METTL3 was significantly upregulated in GISTs tissues and cell lines. Functional experiments demonstrated that overexpression of METTL3 promoted GISTs cell malignant biological behavior and tumor growth in vitro and in vivo, and conversely, silencing of METTL3 had opposite effects and suppressed GISTs progression. Further mechanistical experiments verified that METTL3 promoted the maturation of miR-25-3p in an m6A-dependent manner. Similar to METTL3, miR-25-3p was also validated as an oncogene to promote cancer development in GISTs. Finally, our rescuing experiments hinted that silencing of miR-25-3p abrogated the tumor-initiating effects of METTL3 overexpression on GISTs. CONCLUSION: Collectively, those results indicated that METTL3 played an oncogenic role in GISTs through positively modulating the miR-25-3p in an m6A-dependent manner, and we firstly discussed how the METTL3/m6A/miR-25-3p axis affected GISTs development.
Authors: Andrew P Garner; Joseph M Gozgit; Rana Anjum; Sadanand Vodala; Alexa Schrock; Tianjun Zhou; Cesar Serrano; Grant Eilers; Meijun Zhu; Julia Ketzer; Scott Wardwell; Yaoyu Ning; Youngchul Song; Anna Kohlmann; Frank Wang; Tim Clackson; Michael C Heinrich; Jonathan A Fletcher; Sebastian Bauer; Victor M Rivera Journal: Clin Cancer Res Date: 2014-09-19 Impact factor: 12.531