Reverse transcription-polymerase chain reaction (RT-PCR)

RNA of Hep-2 cells with transfection of si-NC or 2 PRDX3 shRNAs were extracted with Tri-zol reagent (Invitrogen, Grand Island, NY, USA) following the manufacturer's instructions. RNA (1ug) was reverse transcribed with 2 U Avian Myeloblastosis Virus Reverse Transcriptase (Promega, Madison, WI, USA), as described by the manufacturer. The primer sequences for PRDX3 were: forward, 5’-GTTGTCGCAGTCTCAGTGG-3’; reverse, 5’-GACGCTCAAATGCTTGATG-3’. β -actin (forward, 5’-ACGTTGACATCCGAAAGACC-3’; reverse, 5’-CCACCGATCCACACAGAGTA-3’) was used as the internal control. PCR products were analyzed by Gene Tools (Syngene, Frederick, MD, USA). Results were shown as the relative quantity of target genes per β-actin. Deionized water was used in place of cDNA as a negative control.