In this report, we demonstrate the selective blockade of Ascl2 in HT-29 and LS174T cells could inhibit cell growth, invasion and migration in vitro, and lead to growth arrest in vivo, that is partially related to miRNA-302b-related inhibition of ‘stemness’ of colon cancer progenitor cells based on the experiments of shRNA-Ascl2/HT-29 transfected with miRNA-302b mimic. This evidence concerns the gene ASCL2 and malignant colon neoplasm.