DNA microarray analysis following gefinitib/luteolin administration to PC-3 cells

To further investigate the molecular mechanism of cell death induction by luteolin and/or gefitinib, we performed DNA microarray analysis, which may reveal the change of gene expression pattern following treatment with these chemicals. We examined PC-3 cells after 24h of treatment with 60μM luteolin and/or 60μM gefitinib because subG1 population has not appeared yet at this timing, which helps to avoid the secondary effects of apoptosis on the alteration of the gene expression pattern. PC-3 cells were treated for 24h with DMSO, 60μM luteolin (Lut), 60μM gefitinib (Gef) or their co-administration (Lut+Gef). To calculate the fold change, we compared the hybridization signal intensity between time zero (before addition of any reagents to PC-3 cells) and 24h after addition of DMSO, luteolin, gefitinib and luteolin+gefitinib. Each sample was run in duplicate.

为进一步探究木犀草素（luteolin）与/或吉非替尼（gefitinib）诱导细胞死亡的分子机制，我们开展了DNA微阵列（DNA microarray）分析，以揭示上述化合物处理后基因表达谱的变化。我们选取PC-3细胞，分别以60μM木犀草素、60μM吉非替尼或二者联合处理24小时；选择该时间点的原因在于，此时亚G1期细胞群尚未出现，可规避细胞凋亡对基因表达谱改变产生的次级效应。PC-3细胞的处理分组包括：二甲基亚砜（DMSO）处理组、60μM木犀草素（Lut）处理组、60μM吉非替尼（Gef）处理组，以及二者联合给药组（Lut+Gef），所有组的处理时长均为24小时。为计算基因表达的倍数变化，我们对比了零时刻（即向PC-3细胞添加任何试剂前）与添加DMSO、木犀草素、吉非替尼及二者联合试剂后24小时的杂交信号强度。每份样本均进行两次重复实验。