RNA Expression Analysis of A549 and NCI-H460 Cells

The method was as described previously 27 (link),28 (link). Briefly, A549 and NCI-H460 cells were respectively treated with CyH (0, 1.51, 3.13, 6.25, 12.5, and 25 μM) for 16 h. Total RNA in the treated and untreated cells was extracted using Trizol reagent and the mRNA levels of the target genes were analyzed using One Step SYBR^® PrimeScript^® RT-PCR (TaKaRa, China). All primers were synthesized by Sangon Biotech (Shanghai, China) and the sequences were listed in Table 1. The reaction conditions were as follows: 42 °C for 5 min, then 95 °C for 10 s, followed by 40 cycles (95 °C for 5 s and 60 °C for 31 s). β-actin was used as a control to normalize the relative mRNA levels of all target genes.