Oridonin induced autophagy in human cervical carcinoma HeLa cells through Ras, JNK, and P38 regulation
Oridonin induced autophagy in human cervical carcinoma HeLa cells through Ras, JNK, and P38 regulation
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DOI：
10.1254/jphs.fp0070336
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发表时间：
2007-12-01
影响因子：
3.5
通讯作者：
Ikejima, Takashi
中科院分区：
文献类型：
--
作者：
Cui, Qlao;Tashiro, Shin-Ichl;Ikejima, Takashi
在本研究中，我们研究了冬凌草甲素在HeLa细胞中诱导的自体复制。观察冬凌草甲素对HeLa细胞的荧光变化、自噬水平和蛋白表达的影响。冬凌草甲素在体外以剂量和时间依赖的方式诱导HeLa细胞自噬。冬凌草甲素处理的HeLa细胞预先标记了自噬小体特异性染料MDC，吸收了更多的MDC阳性颗粒，并具有明显更高的荧光密度；同时，冬凌草甲素增加了自噬相关蛋白MAP-LC3和Beclin 1的表达。在冬凌草甲素诱导的Hela细胞中，3-甲基腺嘌呤(3-MA)可剂量依赖性地降低自噬比率，同时下调MAP-LC3和Beclin 1的蛋白表达。此外，当应用RAS抑制剂时，自噬水平升高，而P38和JNK抑制剂显著降低自噬比率，表明冬凌草甲素诱导的自噬过程受RAS负调控，但受P38和JNK MAPKs正调控。RAF-1和ERK1/2与这些信号通路无明显相关性。
In this study, we investigated autopliagy induced by orldonin in HeLa cells. HeLa cells were exposed to oridonin, and the fluorescent changes, autophagic levels, and protein expressions were evaluated. Oridonin induced autophagy in HeLa cells in vitro in a dose- and time-dependent manner. Oridonin-treated HeLa cells, which had been prelabeled with the autophagosome-specific dye monodansylcadervarine (MDC), recruited more MDC-positive particles and had a significantly higher fluorescent density; and simultaneously, expressions of autophagy-related proteins, MAP-LC3 and Beclin 1, were increased by oridonin. In oridonin-induced Hela cells, pretreatment with 3-methyladenine (3-MA, the specific inhibitor of autophagy) dose-dependently decreased the autophagic ratio accompanied with downregulation of the protein expressions of MAP-LC3 and Beclin 1. Furthermore, when a Ras inhibitor was applied, the autophagic levels were augmented, whereas P38 and JNK inhibitors decreased the autophagic ratio significantly, indicating that this oridonin-induced autophagic process was negatively regulated by Ras, but positively regulated by P38 and JNK MAPKs. Raf-1 and ERK1/2 had no obvious correlation to these signaling pathways.