Down-regulated FTO and ALKBH5 co-operatively activates FOXO signaling through m6A methylation modification in HK2 mRNA mediated by IGF2BP2 to enhance glycolysis in colorectal cancer

Mujie Ye; Jinhao Chen; Feiyu Lu; Minghui Zhao; Suwen Wu; Chunhua Hu; Ping Yu; Jingbao Kan; Jianan Bai; Ye Tian; Qiyun Tang

doi:10.1186/s13578-023-01100-9

Down-regulated FTO and ALKBH5 co-operatively activates FOXO signaling through m6A methylation modification in HK2 mRNA mediated by IGF2BP2 to enhance glycolysis in colorectal cancer

Cell Biosci. 2023 Aug 14;13(1):148. doi: 10.1186/s13578-023-01100-9.

Authors

Mujie Ye^#¹, Jinhao Chen^#¹, Feiyu Lu¹, Minghui Zhao², Suwen Wu³, Chunhua Hu¹, Ping Yu¹, Jingbao Kan¹, Jianan Bai¹, Ye Tian⁴, Qiyun Tang⁵

Affiliations

¹ Department of Geriatric Gastroenterology, Institute of Neuroendocrine Tumor, Neuroendocrine Tumor Center, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing Medical University, NO.300 Guangzhou Road, Nanjing, 210029, China.
² Department of Radiation Oncology, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
³ Obstetrics and Gynaecology Hospital, Fudan University, Shanghai, China.
⁴ Department of Geriatric Gastroenterology, Institute of Neuroendocrine Tumor, Neuroendocrine Tumor Center, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing Medical University, NO.300 Guangzhou Road, Nanjing, 210029, China. tianye6626@126.com.
⁵ Department of Geriatric Gastroenterology, Institute of Neuroendocrine Tumor, Neuroendocrine Tumor Center, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing Medical University, NO.300 Guangzhou Road, Nanjing, 210029, China. tqy831@163.com.

^# Contributed equally.

Abstract

Background: N6-methyladenosine (m6A) modification is the most abundant reversible methylation modification in eukaryotes, and it is reportedly closely associated with a variety of cancers progression, including colorectal cancer (CRC). This study showed that activated lipid metabolism and glycolysis play vital roles in the occurrence and development of CRC. However, only a few studies have reported the biological mechanisms underlying this connection.

Methods: Protein and mRNA levels of FTO and ALKBH5 were measured using western blot and qRT-PCR. The effects of FTO and ALKBH5 on cell proliferation were examined using CCK-8, colony formation, and EdU assays, and the effects on cell migration and invasion were tested using a transwell assay. m6A RNA immunoprecipitation (MeRIP) and RNA-seq was used to explore downstream target gene. RIP was performed to verify the interaction between m6A and HK2. The function of FTO and ALKBH5 in vivo was determined by xenograft in nude mice.

Results: In this study, FTO and ALKBH5 were significantly down-regulated in CRC patients and cells both in vivo and in vitro in a high-fat environment. Moreover, FTO and ALKBH5 over-expression hampered cell proliferation both in vitro and in vivo. Conversely, FTO and ALKBH5 knockdown accelerated the malignant biological behaviors of CRC cells. The mechanism of action of FTO and ALKBH5 involves joint regulation of HK2, a key enzyme in glycolysis, which was identified by RNA sequencing and MeRIP-seq. Furthermore, reduced expression of FTO and ALKBH5 jointly activated the FOXO signaling pathway, which led to enhanced proliferation ability in CRC cells. IGF2BP2, as a m6A reader, positively regulated HK2 mRNA in m6A dependent manner. Additionally, down-regulation of FTO/ALKBH5 increased METTL3 and decreased METTL14 levels, further promoting CRC progression.

Conclusion: In conclusion, our study revealed the FTO-ALKBH5/IGF2BP2/HK2/FOXO1 axis as a mechanism of aberrant m6A modification and glycolysis regulation in CRC.

Keywords: Colorectal cancer; Glycolysis; N6-methyladenosine; RNA demethylase.

Abstract

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