〔Abstract〕 Objective To investigate the effect of long non-coding RNA(LncRNA) anoctamin 1 antisense RNA-1(ANO1-AS1) on the proliferation and apoptosis of esophageal squamous cell carcinoma(ESCC) cells and its possible mechanisms. Methods Silenced ANO1-AS1 lentivirus was transfected in ESCC cells TE-1 and EC109. Subsequently, the expression levels of ANO1-AS1 and calcium-activated chloride channel protein 1(ANO1) in the cells were detected by qRT-PCR. CCK-8 and colony formation assays were used to detect the proliferation of TE-1 and EC109 cells. ANO1 positively related expressed genes were obtained from the LinkedOmics database and then the gene set was enriched for pathways and possible pathways were validated. The expression levels of proliferating cell nuclear antigen(PCNA), P53 protein, apoptosis-related protein(Bax and Bcl-2), ANO1 protein and phosphatidylinositol-3-kinase/protein kinase B(PI3 K/Akt) pathway-related protein were assessed by Western blot. Results After transfection of lentivirus with silent expression function, the expression level of ANO1-AS1 was significantly reduced in TE-1 and EC109 cells(P<0.05); After down-regulation of ANO1-AS1, compared with the negative control group, the proliferation ability of ESCC cells was reduced(P<0.05) and the rate of clone formation decreased(P<0.05); Western blot results showed that, compared with negative controls, the expression of PCNA decreased, the expression of oncogene P53 protein increased(P<0.05), the expression of proteins(Bax) increased, Bcl-2 decreased and the levels of phosphorylation of the pathway proteins PI3 K and Akt decreased(P<0.05). Conclusion Knockdown of ANO1-AS1 can decrease proliferation and promote apoptosis in ESCC, which may be achieved by affecting PI3 K/Akt pathway activation.