PI-103 and sorafenib inhibit hepatocellular carcinoma cell proliferation by blocking Ras/Raf/MAPK and PI3K/AKT/mTOR pathways.
PI-103 and sorafenib inhibit hepatocellular carcinoma cell proliferation by blocking Ras/Raf/MAPK and PI3K/AKT/mTOR pathways.
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发表时间：
2010-12
影响因子：
2
通讯作者：
R. Gedaly;P. Angulo;J. Hundley;M. Daily;Changguo Chen;A. Koch;B. Evers
中科院分区：
文献类型：
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作者：
R. Gedaly;P. Angulo;J. Hundley;M. Daily;Changguo Chen;A. Koch;B. Evers
背景 在肝细胞癌 (HCC) 中发现异常的 Ras/Raf/MAPK 和 PI3K/AKT/mTOR 信号通路。本研究报告了索拉非尼（一种多激酶抑制剂）和 PI-103（一种双重 PI3K/mTOR 抑制剂）如何单独或联合抑制 HCC 细胞系 Huh7 的增殖。材料和方法 Huh7 增殖通过 3 H-胸苷掺入和 MTT 测定来测定。 Western blot用于检测Ras/Raf和PI3K通路中关键酶的磷酸化。结果 索拉非尼和 PI-103 作为单一药物以剂量依赖性方式抑制 Huh7 增殖和表皮生长因子 (EGF) 刺激的 Huh7 增殖。索拉非尼和 PI-103 的组合产生了协同效应。 EGF 增加 MEK 和 ERK（关键 Ras/Raf 下游信号蛋白）的磷酸化；这种激活被索拉非尼抑制。然而，索拉非尼作为单一药物可增加 AKT(Ser473) 和 mTOR 磷酸化。 PI-103 抑制 EGF 刺激的 PI3K/AKT/mTOR 通路成分的激活。 PI-103 是 AKT(Ser473) 磷酸化的有效抑制剂；相反，雷帕霉素刺激 AKT(Ser473) 磷酸化。研究发现，PI-103 作为单一药物可刺激 MEK 和 ERK 磷酸化。然而，索拉非尼和 PI-103 的组合会抑制 Ras/Raf 和 PI3K 通路中所有测试的激酶。结论 索拉非尼联合 PI-103 可以通过阻断 Ras/Raf/MAPK 和 PI3K/AKT/mTOR 通路，显着抑制 EGF 刺激的 Huh7 增殖。
BACKGROUND Aberrant Ras/Raf/MAPK and PI3K/AKT/mTOR signaling pathways are found in hepatocellular carcinoma (HCC). This study reports how sorafenib (a multi-kinase inhibitor) and PI-103 (a dual PI3K/mTOR inhibitor) alone and in combination inhibit the proliferation of the HCC cell line, Huh7. MATERIALS AND METHODS Huh7 proliferation was assayed by 3H-thymidine incorporation and by MTT assay. Western blot was used to detect phosphorylation of the key enzymes in the Ras/Raf and PI3K pathways. RESULTS Sorafenib and PI-103, as single agents inhibited Huh7 proliferation and epidermal growth factor (EGF)-stimulated Huh7 proliferation in a dose-dependent fashion; the combination of sorafenib and PI-103 produced synergistic effects. EGF increased phosphorylation of MEK and ERK, key Ras/Raf downstream signaling proteins; this activation was inhibited by sorafenib. However, sorafenib as a single agent increased AKT(Ser473) and mTOR phosphorylation. EGF-stimulated activation of PI3K/AKT/mTOR pathway components was inhibited by PI-103. PI-103 is a potent inhibitor of AKT(Ser473) phosphorylation; in contrast, rapamycin stimulated AKT(Ser473) phosphorylation. It was found that PI-103, as a single agent, stimulated MEK and ERK phosphorylation. However, the combination of sorafenib and PI-103 caused inhibition of all the tested kinases in the Ras/Raf and PI3K pathways. CONCLUSION The combination of sorafenib and PI-103 can significantly inhibit EGF-stimulated Huh7 proliferation by blocking both Ras/Raf/MAPK and PI3K/AKT/mTOR pathways.