Grifolin induces autophagic cell death by inhibiting the Akt/mTOR/S6K pathway in human ovarian cancer cells
Grifolin induces autophagic cell death by inhibiting the Akt/mTOR/S6K pathway in human ovarian cancer cells
复制标题
Grifolin 通过抑制人卵巢癌细胞中的 Akt/mTOR/S6K 通路诱导自噬性细胞死亡
DOI：
10.3892/or.2016.4840
复制
发表时间：
2016-08-01
期刊：
影响因子：
4.2
通讯作者：
Jiang, Jie
中科院分区：
文献类型：
--
作者：
Che, Xiaoxia;Yan, Hong;Jiang, Jie
Grifolin是一种分离自Albatrellus confluence的次级代谢产物，已被报道具有多种肿瘤的抗肿瘤活性。到目前为止，还没有关于自噬在Grifolin处理的人卵巢癌细胞中的作用的报道。本研究旨在探讨自噬在卵巢癌中的作用及其机制。卵巢癌细胞系A2780和SKOV 3用grifolin处理。MTT法检测细胞增殖情况，流式细胞仪、电镜、免疫荧光染色和GFP-LC 3斑点形成实验检测细胞自噬作用。蛋白质印迹法检测细胞自噬标志物和主要自噬相关Akt/mTOR/S6 K通路蛋白的表达。MTT法检测表明，grifolin对人卵巢癌细胞株A2780和SKOV 3的增殖有抑制作用。流式细胞术、电镜、免疫荧光和GFP-LC 3斑点形成实验证实了Grifolin诱导人卵巢癌细胞自噬性死亡。蛋白质印迹分析的结果表明，灰树花素处理导致自噬标志物LC 3B、Atg 7、Beclin-1的上调以及P62的下调沿着。此外，p-Akt、p-mTOR、p-p70 S6 K和p-4 E-BP 1途径的蛋白质下调，而这些蛋白质的总量保持不受影响。本研究表明，grifolin可通过抑制Akt/mTOR/S6 K通路诱导人卵巢癌细胞自噬性死亡。
Grifolin, a secondary metabolic product isolated from the mushroom Albatrellus confluence, has been reported to possess antitumor activities in various tumors. To date, no report exists on the role of autophagy in grifolin-treated human ovarian cancer cells. In the present study, we investigated the effect and the mechanism of autophagy in ovarian cancer. Ovarian cancer cell lines A2780 and SKOV3 were treated with grifolin. Cell proliferation was assessed by MTT assay and the autophagic effect was determined using flow cytometry, electron microscopy, immunofluorescence staining and GFP-LC3 puncta formation assay. The expression of autophagy markers and the main autophagy-associated Akt/mTOR/S6K pathway proteins were measured by western blot analysis. MTT assay indicated that grifolin inhibits the proliferation of human ovarian cancer cell lines A2780 and SKOV3. Flow cytometry, electron microscopy, immunofluorescence and GFP-LC3 puncta formation assay proved that grifolin induces autophagic cell death in human ovarian cancer. The results of the western blot analysis suggested that grifolin treatment leads to upregulation of autophagy markers LC3B, Atg7, Beclin-1 along with downregulation of P62. In addition, the proteins of the pathways p-Akt, p-mTOR, p-p70S6K and p-4E-BP1 were downregulated while the total of these proteins remained unaffected. The present study indicated that grifolin could induce autophagic cell death in human ovarian cancer by inhibiting the Akt/mTOR/S6K pathway.