Shaorui Niu,1 Xiaozhe Zheng,1 Yuyang Yao,1 Yue Dong,2 Yupan Hu,3 Zhiyang Xiao,1 Jiaxue Yang,1 Chengli Jiang,1 Xin Zou,4 Zihao Zou,5 Pang Yang1 

1Jiangxi Key Laboratory of Cancer Metastasis and Precision Treatment, Central Laboratory, The First Hospital of Nanchang, The Third Affiliated Hospital of Nanchang University, Nanchang, People’s Republic of China; 2Zhejiang Provincial Clinical Research Center for Obstetrics and Gynecology, Department of Obstetrics and Gynecology, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, People’s Republic of China; 3Fuzhou Medical College of Nanchang University, Fuzhou City, Jiangxi Province, People’s Republic of China; 4Clinical Medical College, Ningxia Medical University, Yinchuan, Ningxia Province, People’s Republic of China; 5Department of Orthopaedic Surgery, Fourth Hospital, Harbin Medical University, Harbin, People’s Republic of China

Correspondence: Pang Yang, Email pyang392@163.com

Background: Although adenosylhomocysteinase (AHCY) is crucial to the oncogenesis and growth of some cancers, it is unknown how this affects bladder urothelial carcinoma (BLCA). Investigating the variations in AHCY expression in BLCA and examining the relationship between AHCY expression and BLCA patient prognosis were the goals of this investigation.
Methods: By leveraging The Cancer Genome Atlas (TCGA) database, we undertook a meticulous examination of AHCY expression levels, juxtaposing them between BLCA and normal tissues. Subsequently, Kaplan-Meier analysis and COX regression and nomogram was used to assess the effect of AHCY on the survival of BLCA patients. We further elaborated on the possible enriched pathways of AHCY and its immune relevance. In addition, we employed si-RNA technology to downregulate the AHCY gene expression and subsequently utilized quantitative real-time PCR (qRT-PCR), CCK-8, cell scratch assays, and Transwell migration assays to validate the pivotal role of AHCY in BLCA.
Results: The expression of AHCY was associated with various types of malignancies (including BCLA). In BLCA cancer tissues, there was an observed upregulation of AHCY expression in comparison to paracancerous tissues. Increased expression of AHCY was linked to decreased overall survival (OS), clinical stage, N stage, and T stage in individuals with BLCA. The functional enrichment of AHCY related genes mainly involves biological processes such as rRNA metabolic processes, proteasome activity, and cell cycle regulation, etc. Furthermore, AHCY showed significant associations with m6A related genes and infiltration of immune cells (Especially for Th2 cells and T-gd lymphocytes). In vitro functional experiments substantiated that the inhibition of AHCY effectively suppresses the growth, migration, and invasion of bladder cancer cells.
Conclusion: This study provides novel insights into the role of AHCY in BLCA, which holds significant potential to contribute towards advancing the diagnosis and treatment of BLCA in the future.

Keywords: adenosylhomocysteinase, bladder urothelial carcinoma, biomarker, immune infiltration, prognosis