qRT-PCR assay

Total RNA was acquired using TRIzol (TaKara, Kusatsu, Japan) following the manufacturer’s instructions. The RNA was transformed into cDNA by reverse transcription using cDNA (Vazyme, Nanjing, China). Subsequently, cDNA was used for amplification. We performed qPCR with SYBR Green PCR kit (Vazyme, Nanjing, China) followed by reference manual. GAPDH was used as endogenous control. Primer sequences for PCR were listed as following:

GAPDH, forward: 5′-CTTTGGTATCGTGGAAGGACTC-3′;

reverse: 5′-GTAGAGGCAGGGATGATGTTCT-3′;

Bcl-2, forward: 5′-GATCCTCGAGATGGCGCACGCTGGGAGAAC-3′;

reverse: 5′-GATCGGATCCTCATGGCTGAGCGCAG-3′;

Bax, forward: 5′-GGACGAACTGGACAGTAACATGG-3′;

reverse: 5′-GCAAAGTAGAA-AAGGGCGACAAC-3′;

E-cadherin, forward, 5′-CGAGAGCTACACGTTCACGG-3′;

reverse: 5′-GGGTGTCGAGGGAAAAATAGG-3′;

N-cadherin, forward, 5′-TTTGATGGAGGTCTCCTAACACC-3′;

reverse: 5′-ACGTTTAACACGTTGGAAATGTG-3′.

The 2−ΔΔCT method (Livak and Schmittgen 2001) was used to quantify relative gene expression.