After lncRNA POU6F2‐AS2 knockdown, tumour shrinkage was observed, and the expression of cancer resistance‐related gene (P‐gp, MRP2 and BRCA2) was down‐regulated, indicating lncRNA POU6F2‐AS2 knockdown enhanced sensitivity of 5‐FU. This evidence concerns the gene POU6F2 and neoplasm.