Involvement of ER stress and activation of apoptotic pathways in fisetin induced cytotoxicity in human melanoma.
Involvement of ER stress and activation of apoptotic pathways in fisetin induced cytotoxicity in human melanoma.
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DOI：
10.1016/j.abb.2014.06.034
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发表时间：
2014-12-01
影响因子：
3.9
通讯作者：
Mukhtar H
中科院分区：
文献类型：
--
作者：
Syed DN;Lall RK;Chamcheu JC;Haidar O;Mukhtar H
尽管恶性黑色素瘤的治疗策略最近取得了进展，但恶性黑色素瘤的预后仍然很差。非瑟酮是一种膳食类黄酮，目前正在研究其在各种癌症模型中的生长抑制特性。我们之前表明非瑟酮在体外和体内抑制黑色素瘤的生长。在这里，我们评估了非瑟酮在转移性人黑色素瘤细胞中诱导细胞毒性的分子基础。内质网应激标记物（包括 IRE1α、XBP1s、ATF4 和 GRP78）上调表明，非瑟酮治疗可在高度侵袭性的 A375 和 451Lu 人黑色素瘤细胞中诱导内质网 (ER) 应激。时程分析表明内质网应激与外源性和内源性细胞凋亡途径的激活有关。漆黄素处理的二维黑色素瘤培养物显示出自噬反应并伴有细胞凋亡的诱导。在 3D 重建黑色素瘤模型中，使用非瑟酮进行长期治疗（16 天）可抑制黑色素瘤进展，并伴有显着的细胞凋亡，这一点可以通过处理的构建体中裂解的 Caspase-3 的染色增加来证明。然而，治疗组和对照组之间自噬标记物 LC-3 的表达没有差异。 Fisetin 对 2-D 黑色素瘤培养物的处理导致多功能 AMPK 激活蛋白激酶 (AMPK) 的磷酸化和激活，该激酶参与多种细胞过程的调节，包括自噬和细胞凋亡。 AMPK 沉默不能阻止细胞死亡，表明非瑟酮诱导的细胞毒性是通过 AMPK 依赖性和非依赖性机制介导的。综上所述，我们的研究证实细胞凋亡是非瑟酮抑制黑色素瘤细胞生长的主要机制，并且外在和内在途径的激活有助于非瑟酮诱导的细胞毒性。
The prognosis of malignant melanoma remains poor in spite of recent advances in therapeutic strategies for the deadly disease. Fisetin, a dietary flavonoid is currently being investigated for its growth inhibitory properties in various cancer models. We previously showed that fisetin inhibited melanoma growth in vitro and in vivo. Here, we evaluated the molecular basis of fisetin induced cytoxicity in metastatic human melanoma cells. Fisetin treatment induced endoplasmic reticulum (ER) stress in highly aggressive A375 and 451Lu human melanoma cells, as revealed by up- regulation of ER stress markers including IRE1α, XBP1s, ATF4 and GRP78. Time course analysis indicated that the ER stress was associated with activation of the extrinsic and intrinsic apoptotic pathways. Fisetin treated 2-D melanoma cultures displayed autophagic response concomitant with induction of apoptosis. Prolonged treatment (16 days) with fisetin in a 3-D reconstituted melanoma model resulted in inhibition of melanoma progression with significant apoptosis, as evidenced by increased staining of cleaved Caspase-3 in the treated constructs. However, no difference in the expression of autophagic marker LC-3 was noted between treated and control groups. Fisetin treatment to 2-D melanoma cultures resulted in phosphorylation and activation of the multifunctional AMPK-activated protein kinase (AMPK) involved in the regulation of diverse cellular processes, including autophagy and apoptosis. Silencing of AMPK failed to prevent cell death indicating that fisetin induced cytotoxicity is mediated through both AMPK-dependent and -independent mechanisms. Taken together, our studies confirm apoptosis as the primary mechanism through which fisetin inhibits melanoma cell growth and that activation of both extrinsic and intrinsic pathways contributes to fisetin induced cytotoxicity.
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影响因子：
11.1
作者：
He C;Klionsky DJ
通讯作者：
Klionsky DJ
影响因子：
9.7
作者：
Jang, Ki Young;Jeong, Soo-Jin;Kim, Sung-Hoon
通讯作者：
Kim, Sung-Hoon
影响因子：
3.9
作者：
Chung, Sangwoon;Yao, Hongwei;Caito, Samuel;Hwang, Jae-Woong;Arunachalam, Gnanapragasam;Rahman, Irfan
通讯作者：
Rahman, Irfan
影响因子：
8.8
作者：
Martin S;Lamb HK;Brady C;Lefkove B;Bonner MY;Thompson P;Lovat PE;Arbiser JL;Hawkins AR;Redfern CP
通讯作者：
Redfern CP
影响因子：
4.8
作者：
Go, YM;Gipp, JJ;Jones, DP
通讯作者：
Jones, DP