The cells were subjected to seeding into 6‐well plates containing 1 mL Ham's F12 medium before transfection. After 18 hours, expression plasmids pCMV6‐XL5‐CDX2 (CDX2) and pCMV6‐XL5‐HEPH (HEPH) (OriGene Technologies, Rockville, MD, USA) and small interfering RNAs (siRNA) targeting CDX2 (si‐CDX2) and HEPH (si‐HEPH) were used for transfection, which was conducted according to the manufacturer's instructions of LipofectamineTM 2000 (Invitrogen, Carlsbad, CA, USA). In order to monitor the overexpression of CDX2 and HEPH, mRNA level or protein level was measured after 24 or 48 hours, respectively. miR‐24‐3p mimic, inhibitor and their controls were produced by GENECHEM (Shanghai, China) and transfected into cells using Lipofectamine 2000. MTX (Sigma, Sigma‐Aldrich, St. Louis, MO, USA) was added to cells after 24‐h transfection to analyse the sensitivity of the cells to MTX. The cell viability was assessed by cell counting kit (CCK)‐8 assay. Empty vector (pCMV6‐XL5) and si‐negative control (NC) were used as NC.

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