Impact of ASCL2 on prognosis and cellular biological behaviors of gastric cancer based on bioinformatics analysis and experimental investigation

YANG Shuai; FENG Qingqing; ZHAO Wenwen; ZHU Chao; XIAO Jun; WEI Hongmei

doi:10.3969/j.issn.1009-0460.2025.04.001

Abstract：Objective To analyze the relationship between achaete-scute family bHLH transcription factor 2(ASCL2)expression and poor prognosis in gastric cancer patients through bioinformatics,and to validate the impact of ASCL2 expression on the cellular biological behaviors of gastric cancer cells using cellular experiments.Methods Expression of ASCL2 in pan-cancer and gastric cancer was analyzed using the TIMER 2.0 database.The Kaplan-Meier Plotter database was utilized to assess the influence of ASCL2 expression on the prognosis of gastric cancer patients.Relationships of ASCL2 expression with clinical characteristics,immune cell infiltration,immune checkpoint genes and ESTIMATE scores were analyzed using the UALCAN,TIMER 2.0 and Sangerbox database.Gastric cancer cells with significantly high ASCL2 expression were selected,and the expression of ASCL2 was downregulated through transfection of small interfering RNA.CCK-8,flow cytometry,Transwell,and wound healing assays were conducted to verify the effects of the ASCL2 gene on gastric cancer cell proliferation,apoptosis,invasion,and migration.Results A total of 63 differentially expressed genes related to distant metastasis in gastric cancer were identified from the TCGA database.Compared with adjacent normal tissues,ASCL2 was significantly overexpressed in gastric cancer(P<0.001).High ASCL2 expression was significantly associated with shorter overall survival,first progression,and post-progression survival in gastric cancer patients(P<0.0001).ASCL2 expression was significantly correlated with gastric cancer grade,stage,and lymph node metastasis(P<0.05).ASCL2 expression was negatively correlated with the infiltration levels of CD8+T cells,neutrophils,monocytes,macrophages,and dendritic cells(P<0.05),but not with CD4+T cell infiltration levels(P>0.05).Using the Sangerbox database,a correlation was found between ASCL2 expression and 45 of 60 immune checkpoint pathway genes(P<0.05).Additionally,ASCL2 expression was negatively correlated with stromal scores,immune scores,and ESTIMATE scores.Knockdown of ASCL2 expression inhibited gastric cancer cell proliferation,migration,and invasion,while promoting apoptosis.Conclusion High expression of ASCL2 in gastric cancer may be associated with poor prognosis,suggesting its potential significance as a prognostic indicator and demonstrating its potential role in the pathogenesis and progression of gastric cancer.