Luteolin and gefitinib regulation of EGF signaling pathway and cell cycle pathway genes in PC-3 human prostate cancer cells.
Luteolin and gefitinib regulation of EGF signaling pathway and cell cycle pathway genes in PC-3 human prostate cancer cells.
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DOI：
10.1016/j.jsbmb.2010.06.006
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发表时间：
2010-10
影响因子：
4.1
通讯作者：
Rodriguez, Mary
中科院分区：
文献类型：
--
作者：
Markaverich, Barry M.;Vijjeswarapu, Mary;Shoulars, Kevin;Rodriguez, Mary
关键词：
我们实验室的 cRNA 微阵列和实时 PCR (qPCR) 研究确定了五个细胞周期途径 (CCP) 基因（CCNA2、CCNE2、CDC25A、CDKN1B、PLK-1）作为 PC-3 前列腺癌细胞中木犀草素的靶标（Shoulars 等人，J Steroid Biochem Mol Biol，118：41-50，2010）。在本文中，对微阵列数据的 Ingenuity Pathway Analysis 鉴定出表皮生长因子信号通路 (EGFSP) 中的 7 个木犀草素调节基因（EGFR、c-Fos、SOS、GRB2、JNK1、MKK4 和 RasGAP），可能参与木犀草素对 CCP 基因和细胞增殖的调节。为了解决这些可能性，我们通过实时 PCR (qPCR) 和蛋白质印迹分析比较了这些 EGFSP 和 CCP 基因与木犀草素和吉非替尼的反应谱（RNA 和蛋白质）。木犀草素和吉非替尼是已知的 EGFR 相关酪氨酸蛋白激酶拮抗剂。因此，如果两种途径中的基因都通过这种共同的作用机制控制，则 EGFR 调节的 EGFSP 或 CCP 基因的反应谱应该非常相似。用木犀草素处理 PC-3 细胞 24 小时，引起 c-Fos 基因表达的 4 倍刺激、CCP 基因的显着抑制 (p<0.001) 和 G2/M 停滞。用吉非替尼处理 PC-3 细胞也以类似于木犀草素的方式抑制大多数 CCP 基因，然而，EGFR 拮抗剂抑制 c-Fos 基因表达，刺激 CDKN1B (p27) 并将细胞阻滞在 G0/G1 期。因此，尽管大多数 CCP 基因对木犀草素或吉非替尼的反应模式相似，但这两种化合物对 EGFSP 基因表达和细胞周期停滞的影响明显不同。组合研究表明，EGFSP 基因对木犀草素的反应不受吉非替尼的影响，尽管这两种化合物在抑制 CCNA2、CCNE2、CDC25A 和 PCNA 的能力方面具有相加性。这些发现表明，木犀草素和吉非替尼通过涉及 EGFR 相关酪氨酸激酶的共同机制调节 CCP 基因表达。相反，木犀草素通过 EGFR-酪氨酸激酶独立机制调节 PC-3 细胞增殖，可能涉及通过组蛋白 H4 结合相互作用对 EGFSP 基因表达进行表观遗传控制，从而导致 c-FOS 和 p21 基因表达上调。
cRNA microarray and real-time PCR (qPCR) studies from our lab identified five Cell Cycle Pathway (CCP) genes (CCNA2, CCNE2, CDC25A, CDKN1B, PLK-1) as targets for luteolin in PC-3 prostate cancer cells (Shoulars et. al, J Steroid Biochem Mol Biol, 118: 41–50, 2010). In this paper, Ingenuity Pathway Analysis of the microarray data identified 7 luteolin-regulated genes (EGFR, c-Fos, SOS, GRB2, JNK1, MKK4 and RasGAP) in the Epidermal Growth Factor Signaling Pathway (EGFSP) potentially involved in luteolin regulation of CCP genes and cell proliferation. To address these possibilities, we compared the response profiles (RNA and protein) of these EGFSP and CCP genes to luteolin and gefitinib by real-time PCR (qPCR) and Western blot analyses. Luteolin and gefitinib are known antagonists of EGFR-associated tyrosine protein kinase. Thus, the response profiles of EGFR regulated EGFSP or CCP genes should be very similar if genes in both pathways are controlled through this common mechanism of action. Treatment of PC-3 cell with luteolin for 24 hours caused a 4-fold stimulation of c-Fos gene expression, significant inhibition (p<0.001) of the CCP genes and G2/M arrest. Treatment of PC-3 cells with gefitinib also inhibited most of the CCP genes in a fashion similar to that of luteolin, however, the EGFR antagonist inhibited c-Fos gene expression, stimulated CDKN1B (p27) and arrested the cells in G0/G1. Thus, although the response patterns of most of the CCP genes to luteolin or gefitinib were similar, the effects of the two compounds on EGFSP gene expression and cell cycle arrest were clearly different. Combination studies revealed that the response of EGFSP genes to luteolin was not affected by gefitinib, even though the two compounds were additive with respect to their abilities to inhibit CCNA2, CCNE2, CDC25A and PCNA. These findings suggest that luteolin and gefitinib regulate CCP gene expression through a common mechanism involving EGFR-associated tyrosine kinase. Conversely, luteolin regulates PC-3 cell proliferation through an EGFR-tyrosine kinase independent mechanism(s), likely involving the epigenetic control of gene EGFSP gene expression through histone H4 binding interactions resulting in the upregulation of c-FOS and p21 gene expression.
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DOI：
10.1073/pnas.011404098
发表时间：
2001-01-02
影响因子：
11.1
作者：
Li, C;Wong, WH
通讯作者：
Wong, WH
影响因子：
3.5
作者：
CARBONE, A;RANELLETTI, FO;CAPELLI, A
通讯作者：
CAPELLI, A
DOI：
10.1016/0022-4731(89)90234-3
发表时间：
1989-11-01
影响因子：
4.1
作者：
MARKAVERICH, BM;GREGORY, RR;MIDDLEDITCH, BS
通讯作者：
MIDDLEDITCH, BS
木犀草素抑制前列腺癌细胞中的胰岛素样生长因子 1 受体信号传导
影响因子：
4.7
作者：
Fang, Jing;Zhou, Qiong;Jiang, Bing-hua
通讯作者：
Jiang, Bing-hua
影响因子：
3.1
作者：
ADLERCREUTZ, H;MOUSAVI, Y;HOCKERSTEDT, K
通讯作者：
HOCKERSTEDT, K