The differential susceptibilities of MCF-7 and MDA-MB-231 cells to the cytotoxic effects of curcumin are associated with the PI3K/Akt-SKP2-Cip/Kips pathway.
The differential susceptibilities of MCF-7 and MDA-MB-231 cells to the cytotoxic effects of curcumin are associated with the PI3K/Akt-SKP2-Cip/Kips pathway.
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MCF-7 和 MDA-MB-231 细胞对姜黄素细胞毒性作用的不同敏感性与 PI3K/Akt-SKP2-Cip/Kips 通路相关
DOI：
10.1186/s12935-014-0126-4
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发表时间：
2014
影响因子：
5.8
通讯作者：
Zhao Z
中科院分区：
文献类型：
--
作者：
Jia T;Zhang L;Duan Y;Zhang M;Wang G;Zhang J;Zhao Z
然而，姜黄素在不同类型癌症中不同细胞毒性的机制仍不清楚。本研究的目的是检测姜黄素对两种不同的乳腺癌细胞MCF-7和MDA-MB-231的浓度和时间相关效应，并研究姜黄素处理诱导的功能变化，以及它们与PI 3 K/Akt-SKP 2-Cip/Kips通路的关系。采用WST-1和克隆形成试验检测姜黄素对乳腺癌细胞（MCF-7）和乳腺癌细胞（MDA-MB-231）的细胞毒性。用QRT PCR和Western Blotting检测CDK相互作用蛋白/激酶抑制蛋白（Cip/Kips）成员（p27、p21和p57）和S期激酶相关蛋白2（SKP 2）的表达。用Western Blotting方法研究姜黄素对磷脂酰肌醇3-激酶（PI 3 K）/Akt及其底物Foxo 1和Foxo 3a的影响。利用靶向SKP 2的小干扰RNA（siRNA）研究SKP 2与Cip/Kips成员的关系。最后，采用WST-1法研究姜黄素对MCF-7和MDA-MB-231细胞的作用以及PKB和SKP 2信号通路对姜黄素敏感性的抑制作用（MDA-MB-231细胞是敏感的，而MCF-7细胞是抗性的），发现这与姜黄素介导的SKP 2-Cip/Kips（p21和p27而不是p57）信号传导的差异调节有关。不同的细胞反应进一步与姜黄素对PI 3 K/Akt及其底物Foxo 1和Foxo 3a的匡威作用有关。重要的是，PI 3 K抑制剂wortmannin可以抵消姜黄素诱导的MCF-7细胞Akt磷酸化和SKP 2上调。随后的WST-1试验表明，姜黄素和渥曼青霉素或SKP 2 siRNA的联合治疗不仅进一步增强了MDA-MB-231细胞对姜黄素的敏感性，而且克服了MCF-7细胞对姜黄素的耐药性。Kips信号通路参与姜黄素的作用机制，并揭示姜黄素对该通路的不一致调节是导致姜黄素的作用机制的原因。这两种细胞类型对姜黄素的不同亲和性。
BackgroundThe mechanism underlying the differential cytotoxicity of curcumin in various cancer types, however, remains largely unclear. The aims of this study is to examine the concentration- and time-related effects of curcumin on two different breast cancer cells, MCF-7 and MDA-MB-231, and investigated the functional changes induced by curcumin treatment, as well as their relationship to the PI3K/Akt-SKP2-Cip/Kips pathway.MethodsFirst, WST-1 and clonogenic assay were performed to determine the cytotoxicity of curcumin in MCF-7 and MDA-MB-231 cells. Then, the expression of CDK interacting protein/Kinase inhibitory protein (Cip/Kips) members (p27, p21 and p57) and S-phase kinase-associated protein-2 (SKP2) was investigated by QRT PCR and Western Blotting. Curcumin’s effect on PI3K (phosphatidylinositol 3-kinase) /Akt and its substrates Foxo1 and Foxo3a were then studied by Western Blotting. Small interfering RNAs (siRNAs) targeting SKP2 was used to explore the relationship between SKP2 and Cip/Kips members. Finally, WST-1 assay was tested to explore the concomitant treatment with curcumin and the inhibition of PKB or SKP2 signaling on curcumin sensitivity in MCF-7 and MDA-MB-231 cells.ResultsWe demonstrated MCF-7 and MDA-MB-231 cells exhibited differential responses to curcumin by WST-1 and clonogenic assay (MDA-MB-231 cells was sensitive, and MCF-7 cells was resistant), which were found to be related to the differential curcumin-mediated regulation of SKP2-Cip/Kips (p21 and p27 but not p57) signaling. The differential cellular responses were further linked to the converse effects of curcumin on PI3K/Akt and its substrates Foxo1 and Foxo3a. Importantly, PI3K inhibitor wortmannin could counteract both curcumin-induced phosphorylation of Akt and up-regulation of SKP2 in MCF-7 cells. Subsequent WST-1 assay demonstrated concomitant treatment with curcumin and wortmannin or SKP2 siRNA not only further augmented curcumin sensitivity in MDA-MB-231 cells but also overcame curcumin resistance in MCF-7 cells.ConclusionsOur study established PI3K/Akt-SKP2-Cip/Kips signaling pathway is involved in the mechanism of action of curcumin and revealed that the discrepant modulation of this pathway by curcumin is responsible for the differential susceptibilities of these two cell types to curcumin.
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DOI：
10.4161/cc.9.5.10914
发表时间：
2010-03-01
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Bashir T;Pagan JK;Busino L;Pagano M
通讯作者：
Pagano M
影响因子：
2.3
作者：
Mehta, K;Pantazis, P;Aggarwal, BB
通讯作者：
Aggarwal, BB
影响因子：
5.8
作者：
Aggarwal, Bharat B.;Banerjee, Sanjeev;Sethi, Gautam
通讯作者：
Sethi, Gautam
影响因子：
1.8
作者：
Commandeur, JNM;Vermeulen, NPE
通讯作者：
Vermeulen, NPE
影响因子：
5.3
作者：
Brunet, A;Park, J;Greenberg, ME
通讯作者：
Greenberg, ME