Laboratory of Biochemistry, National Institute for Digestive Diseases, IRCCS Saverio de Bellis, Castellana Grotte, Italy.
J Cell Physiol. 2013 Feb;228(2):292-7. doi: 10.1002/jcp.24148.
To evaluate the growth-inhibitory properties of the potent multi-kinase antagonist Regorafenib (Fluoro-Sorafenib), which was synthesized as a more potent Sorafenib, a Raf inhibitor and to determine whether similar mechanisms were involved, human hepatoma cell lines were grown in the presence or absence of Regorafanib and examined for growth inhibition. Western blots were performed for Raf targets, apoptosis, and autophagy. Regorafenib inhibited growth of human Hep3B, PLC/PRF/5, and HepG2 cells in a concentration- and time-dependent manner. Multiple signaling pathways were altered, including MAP kinases phospho-ERK and phospho-JNK and its target phospho-c-Jun. There was evidence for apoptosis by FACS, cleavage of caspases and increased Bax levels; as well as induction of autophagy, as judged by increased Beclin-1 and LC3 (II) levels. Prolonged drug exposure resulted in cell quiescence. Full growth recovery occurred after drug removal, unlike with doxorubicin chemotherapy. Regorafenib is a potent inhibitor of cell growth. Cells surviving Regorafenib treatment remain viable, but quiescent and capable of regrowth following drug removal. The reversibility of tumor cell growth suppression after drug removal may have clinical implications.
为了评估强效多激酶抑制剂瑞戈非尼（氟代索拉非尼）的生长抑制特性，该药物是作为更有效的索拉非尼（一种 Raf 抑制剂）合成的，以确定是否涉及类似的机制，在存在或不存在瑞戈非尼的情况下培养人肝癌细胞系，并检查其生长抑制情况。进行了 Western blot 以检测 Raf 靶标、细胞凋亡和自噬。瑞戈非尼以浓度和时间依赖性方式抑制人 Hep3B、PLC/PRF/5 和 HepG2 细胞的生长。多种信号通路发生改变，包括 MAP 激酶磷酸化 ERK 和磷酸化 JNK 及其靶标磷酸化 c-Jun。通过 FACS、半胱天冬酶的切割和 Bax 水平的增加证明了细胞凋亡；以及自噬的诱导，通过增加 Beclin-1 和 LC3（II）水平来判断。延长药物暴露会导致细胞静止。与多柔比星化疗不同，药物去除后会完全恢复生长。瑞戈非尼是一种有效的细胞生长抑制剂。在瑞戈非尼治疗后存活的细胞仍然存活，但处于静止状态，并能够在药物去除后重新生长。药物去除后肿瘤细胞生长抑制的可逆性可能具有临床意义。
