Abstract: The aim of this study was to reveal the effects of anchoring filament protein gene ladinin-1（LAD1） on the proliferation, migration, invasion and apoptosis of esophageal squamous carcinoma（ESCC） cells and its possible mechanisms of action. Transcriptome sequencing of ESCC tissues and paracancerous tissues combined with bioinformatics database analysis revealed that LAD1 was highly expressed in ESCC tissues, and the expression of LAD1 in ESCC cell lines was verified by RT-qPCR. Cancerous tissues and correspon-ding paracancerous tissue specimens from 31 ESCC patients were collected for immunohistochemical staining to detect the expression of LAD1 in ESCC tissues and analyze the correlation between LAD1 and histopathological features of ESCC. siRNA was used to knock down LAD1 expression in KYSE-30 and KYSE-150 cells, and functional changes in ESCC cells were detected by CCK-8 assay, plate cloning assay, scratch healing assay, Transwell assay, and flow cytometry. Upstream transcription factors（TFs） that may regulate LAD1 were predicted on the UCSC and JASPAR databases, and their expression in ESCC cells and tissues was verified by RT-qPCR and immunohistochemical staining. Finally, the regulatory relationship between TFs and LAD1 was verified by using RT-qPCR assay and dual-luciferase assay.The results showed that LAD1 was highly expressed in KYSE-30 and KYSE-150 cells compared to HET-1A cells（P＜0.01 or P＜0.001）, consistent with the transcriptome sequencing results. Immunohistochemical results showed that LAD1 was highly expressed in ESCC and negatively correlated with the degree of histopathological differentiation of ESCC（P＜0.01 or P＜0.001）. Knockdown of LAD1 suppressed ESCC proliferation, migration, invasion and promoted apoptosis（P＜0.01 or P＜0.001）. UCSC and JASPAR database predictions combined with RT-qPCR experiments and immunohistochemical staining speculated that KLF5, EHF, and ELF3 might be upstream transcription factors of LAD1, and RT-qPCR experiments and dual-luciferase experiments were utilized to further verify that LAD1 might be regulated by KLF5, EHF, and ELF3. This study suggests that LAD1 is highly expressed in ESCC cells and tissues and correlates with the degree of pathological differentiation of ESCC tissues; knockdown of LAD1 inhibits the proli-feration, migration, and invasive ability of ESCC cells and promotes apoptosis; KLF5, EHF, and ELF3 are the upstream transcription factors regulating LAD1.