Long non‑coding RNAs (lncRNAs) are widely studied in cancer pathogenesis. Accumulating evidence has demonstrated that lncRNAs are involved in the cellular progression of colorectal cancer (CRC). However, the regulatory mechanism of lncRNA TMPO‑antisense (AS)1 in CRC has not been fully elucidated. The present study aimed to elucidate the role and regulatory mechanisms of lncRNA TMPO‑AS1 in CRC. In the present study, the expression levels of TMPO‑AS1 and microRNA‑143‑3p (miR‑143‑3p) were detected using reverse transcription‑quantitative PCR assay. The relative protein expression levels were measured via western blot analysis. MTT and Transwell assays were used to determine cell proliferation, migration and invasion, while a luciferase reporter assay was performed to assess the relationship between TMPO‑AS1 and miR‑143‑3p. In addition, a tumor animal model was used to investigate the effect of TMPO‑AS1 on tumor growth in CRC in vivo. TMPO‑AS1 expression was increased and miR‑143‑3p expression was decreased in CRC cells. TMPO‑AS1 knockdown and miR‑143‑3p overexpression significantly inhibited cell proliferation, migration and invasion of CRC cells. Luciferase reporter assay results demonstrated that miR‑143‑3p was a direct target of TMPO‑AS1. Inhibition of miR‑143‑3p could alleviate the suppressive effects of TMPO‑AS1 deletion on cell proliferation, migration and invasion of CRC cells. Furthermore, TMPO‑AS1 deletion could inhibit tumor growth in CRC in vivo. It was concluded that TMPO‑AS1 regulated cell proliferation, migration and invasion of CRC cells by targeting miR‑143‑3p. These findings provided a new regulatory network and therapeutic target for the treatment of CRC.