Figure 3.

Quantitative RT-PCR analysis of DAPK and E-cadherin mRNA levels in ESCC and adjacent normal tissues. Total RNA was harvested from tissues using the RNeasy FFPE kit (Qiagen) according to the manufacturer’s instructions. The RT-PCR experiments were repeated at least 3 times. RNA was reverse transcribed into cDNA using random primers in a Reverse Transcription II system (Promega) according to the manufacturer’s instructions. Expression of DAPK and E-cadherin mRNAs was quantified by quantitative PCR using an ABI Prism Sequence Detection System (Applied Biosystems). Template-negative and RT-negative conditions were used as controls. Amplification of the endogenous GAPDH cDNA was monitored. Levels (mean values) of DAPK and E-cadherin transcripts in patients were calculated. RT-PCR, reverse transcription-PCR; ESCC, esophageal squamous cell carcinoma; DAPK, death-associated protein kinase.