m6A demethylase ALKBH5 maintains stemness of intrahepatic cholangiocarcinoma by sustaining BUB1B expression and cell proliferation

Yuan Gao; Miao Yu; Zengyuan Liu; Yi Liu; Zhijun Kong; Chunfu Zhu; Xihu Qin; Yan Li; Liming Tang

doi:10.1016/j.tranon.2023.101858

m⁶A demethylase ALKBH5 maintains stemness of intrahepatic cholangiocarcinoma by sustaining BUB1B expression and cell proliferation

Transl Oncol. 2024 Mar:41:101858. doi: 10.1016/j.tranon.2023.101858. Epub 2024 Jan 18.

Authors

Yuan Gao¹, Miao Yu², Zengyuan Liu³, Yi Liu⁴, Zhijun Kong⁴, Chunfu Zhu⁴, Xihu Qin⁴, Yan Li⁵, Liming Tang⁶

Affiliations

¹ The Institute of Hepatobiliary and pancreatic diseases, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou 213000, China; Department of Hepato-biliary-pancreatic Surgery, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou 213000, China.
² Department of Bioinformatics, Nanjing Medical University, Nanjing 211166, China.
³ The Third People's Provincial Hospital of Henan Province, Zhengzhou, 450000, Henan, China.
⁴ Department of Hepato-biliary-pancreatic Surgery, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou 213000, China.
⁵ Department of Bioinformatics, Nanjing Medical University, Nanjing 211166, China. Electronic address: liyan@njmu.edu.cn.
⁶ Gastrointestinal Surgery and Central Laboratory, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou 213000, China. Electronic address: tangliming@njmu.edu.cn.

Abstract

ALKBH5 plays critical roles in various cellular processes via post-transcriptional regulation of oncogenes or tumor suppressors in an N6-methyladenosine (m6A)-dependent manner. However, its function in intrahepatic cholangiocarcinoma (ICC) remains unclear. In the present study, bioinformatic analyses of The Cancer Genome Atlas (TCGA) data were performed, and the association of ALKBH5 in predicting overall survival in patients with ICC was investigated. Then, the clinical data of patients from The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University (Changzhou, China) was used to reveal the overall survival of patients with ICC with different ALKBH5 expression levels by Kaplan-Meier survival analysis. Subsequently, in vitro and in vivo studies were conducted to explore and verify the downstream genes regulated by ALKBH5. The results from TCGA data demonstrated that ALKBH5 expression is elevated in ICC and that patients with high ALKBH5 expression exhibited poor survival compared with patients with low expression. In addition, in vitro assays demonstrated that ALKBH5 promoted cell viability and maintained the stemness of ICC cells, leading to ICC progression. The present study also demonstrated that BUB1 mitotic checkpoint serine/threonine kinase B (BUB1B) is the downstream gene regulated by ALKBH5 and targeting BUB1B suppressed cell growth. The in vitro and vivo experiments revealed that ALKBH5 might function through BUB1B to maintain the stemness of ICC and that altering BUB1B may suppress ICC progression.

Keywords: ALKBH5; BUB1B; Cancer cell stemness; ICC.