Comprehensive qRT-PCR Analysis of Apoptosis-Related Genes

RT-PCR was performed according to the methods descirbed in a previous study (11 (link)). The RNA of lung tissue was treated with TRIzol reagent (Invitrogen Life Technologies, Carlsbad, CA, USA) for extraction. After total RNA was digested with RNase-free DNase (Roche, Diagnostics Basel, Switzerland) for 15 min at 37°C, then the digested RNA was purified by RNeasy kit (Qiagen, Hilden, Germany). The cDNA was synthesized from 2 μg of total RNA at 37°C for l h with AMV reverse transcriptase (GE Healthcare, Little Chalfont, Buckinghamshire, UK) (2 (link)). Sequences of Bax, Bcl-2, MMPs and TIMPs were specifically amplified (Table I) by thermal cycler (Eppendorf, Hamburg, Germany). The PCR products were separated in 1.0% agarose gels and visualized with ethidium bromide staining.