Objective: To investigate the role of the esophageal squamous cancer-specific ultra-conserved cancer/testis (CT)-LINC01424 in esophageal squamous cancer progression. Methods: The TCGA (The Cancer Genome Atlas) and GTEx (Genotype-Tissue Expression) databases were used to screen the long non-coding RNA (lncRNA) with ultraconservative CT properties specific to esophageal squamous carcinoma, and the nucleoplasmic isolation assay and rapid amplification of cDNA end (RACE) were used for the cellular localization and sequence identification of LINC01424.118 pairs of esophageal cancer samples was used for the analysis of prognosis. After knocking down LINC01424 in Eca109 cell line or overexpressing LINC01424 in KYSE410 cell line, the effects of CT-LINC01424 on the esophageal squamous carcinoma was detected by CCK-8 assay, colony assay, Transwell assay, and nude micel assay; further application of MEM (Multi Experiment Matrix) database for downstream gene exploration. Results: An ultraconservative CT-LINC01424 specific for esophageal squamous carcinoma was identified. Analysis of clinical data showed that CT-LINC01424 was significantly associated with poor clinical stage and prognosis of esophageal squamous carcinoma. Knockdown of the expression of LINC01424 inhibited the proliferation and invasion of esophageal squamous carcinoma cells, while overexpression of LINC01424 resulted in the opposite result. Furthermore, the expression of LINC01424 was significantly co-expressed with KNTC1 (Kinetochore Associated 1) gene. Conclusion: LINC01424 is highly expressed in esophageal squamous carcinoma tissues, promotes the proliferation and invasion of esophageal squamous carcinoma cells, and may exert its pro-carcinogenic effects by affecting the expression of KNTC1.