Qinqing Wu ¹ Lu Ye ² Yuwei Wu ³ Mengyu Zhao ⁴ Jiaxin Lu ² Yanping Yu ⁵ Yixiao Niu ⁵ Luxiao Zhang ² Peijun Zuo ^5*

• ¹ College of Medicine, Shantou University, Shantou, Guangdong Province, China
• ² Chinese Academy of Medical Sciences and Peking Union Medical College, Dongcheng, Beijing Municipality, China
• ³ Fujian Medical University Union Hospital, Fuzhou, Fujian Province, China
• ⁴ First Affiliated Hospital of Anhui Medical University, Hefei, Anhui Province, China
• ⁵ School of Public Health, Shantou University, Shantou, Guangdong Province, China

Background: Colon adenocarcinoma (COAD) is a malignancy with a high mortality rate and complex biological characteristics and heterogeneity, which poses challenges for clinical treatment. Anoikis is a type of programmed cell death that occurs when cells lose their attachment to the extracellular matrix (ECM), and it plays a crucial role in tumor metastasis. However, the specific biological link between anoikis and COAD, as well as its mechanisms in tumor progression, remains unclear, making it a potential new direction for therapeutic strategy research.We employed transcriptomic data and clinical information from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) to pinpoint differentially expressed anoikis-related genes (ARGs) in COAD. Using Cox proportional hazards models and Lasso regression analysis, we developed a prognostic signature derived from these ARGs. We also investigated the roles and interactions of these genes in the tumor microenvironment by analyzing single-cell RNA sequencing data. Additionally, we employed molecular docking techniques to evaluate the potential of inhibin subunit beta B (INHBB) as therapeutic targets and to assess the binding affinity of candidate drugs. Finally, we used gene knockout techniques to silence the key gene INHBB and explored its biological functions in vitro.Results: In our study, by analyzing the expression differences of ARGs, we successfully classified patients with COAD. Kaplan-Meier survival analysis demonstrated that patients with elevated risk scores experienced poorer prognosis, a finding that was confirmed in both the training and validation cohorts. Additionally, immune infiltration analysis revealed a notable increase in immune cell presence within the tumor microenvironment of high-risk patients. Molecular docking identified potential drug candidates with high binding affinity to INHBB, including risperidone. Furthermore, in vitro experiments with INHBB showed that downregulation of its expression in COAD cell lines significantly reduced cellular viability and migration capacity.In summary, our research, based on the expression characteristics of ARGs, provides new insights into the precise classification, prognosis assessment, and identification of potential therapeutic targets in COAD. It also validates the key role of INHBB in the progression of COAD, establishing the foundation for future personalized treatment strategies.