Midkine downregulation increases the efficacy of quercetin on prostate cancer stem cell survival and migration through PI3K/AKT and MAPK/ERK pathway
Midkine downregulation increases the efficacy of quercetin on prostate cancer stem cell survival and migration through PI3K/AKT and MAPK/ERK pathway
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DOI：
10.1016/j.biopha.2018.08.061
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发表时间：
2018-11-01
影响因子：
7.5
通讯作者：
Oktem, Gulperi
中科院分区：
文献类型：
--
作者：
Erdogan, Suat;Turkekul, Kader;Oktem, Gulperi
目的：探讨生长因子中期因子（MK）和类黄酮槲皮素对前列腺癌（PCa）干细胞（CSC）存活、凋亡和迁移的作用。主要方法：采用磁激活细胞分选系统从PC3和LNCaP细胞中分别分离CD44（+）/CD133（+）和CD44（+）干细胞。使用 3D 细胞培养来评估槲皮素、MK siRNA 以及两者的组合抑制球状体形成、细胞凋亡和细胞周期停滞的能力。进行了基于图像的细胞仪、RT-qPCR、Western blotting 和 Transwell 迁移测定。主要发现：槲皮素处理 24-72 小时以时间和剂量依赖性方式抑制 PC3 和 CD44+/CD133+ 干细胞增殖。内源性MK表达的敲低显着抑制CD44(+)/CD133(+)和CD44(+)细胞及其亲代细胞的增殖。 MK siRNA 和槲皮素联合给药比单独治疗更有效地降低细胞存活率、诱导细胞凋亡并引起 G1 期细胞周期停滞。 MK的敲低显着增强了槲皮素对CD44(+)/CD133(+)迁移和球体形成的抑制作用。此外，联合治疗还抑制了PI3K、AKT和ERK1/2的磷酸化，并降低了p38、ABCG2和NF-kappa B的蛋白表达。意义：单独使用槲皮素对CD44(+)/CD133(+)表现出显着的细胞毒作用。 MK在CD44(+)/CD133(+)和CD44(+)细胞的增殖中发挥着重要作用，槲皮素和MK沉默疗法可能是针对在复发、迁移和耐药中发挥作用的CSC的重要策略。
Aims: To examine the functions of growth factor midkine (MK) and a flavonoid quercetin on survival, apoptosis and migration of prostate cancer (PCa) stem cells (CSCs).Main methods: CD44(+)/CD133(+) and CD44(+) stem cells were isolated from PC3 and LNCaP cells, respectively by magnetic-activated cell sorting system. 3D cell culture was used to evaluate the ability of quercetin, MK siRNA, and the combination of both to inhibit spheroid formation, apoptosis and cell cycle arrest. Image-based cytometer, RT-qPCR, Western blotting and transwell migration assays were performed.Key findings: Quercetin treatment for 24-72 h inhibited PC3 and CD44+/CD133+ stem cell proliferation in a time-and dose-dependent manner. Knockdown of endogenous MK expression significantly suppressed proliferation of CD44(+)/CD133(+) and CD44(+) cells as well as their parent cells. Co-administration of MK siRNA and quercetin reduced the cell survival, induced apoptosis and caused G1 phase cell cycle arrest more effectively than the individual therapy. Knockdown of MK significantly enhanced the inhibitory effect of quercetin on CD44(+)/CD133(+) migration and spheroid formation. In addition, the combined therapy inhibited the phosphorylation of PI3K, AKT and ERK1/2, and reduced the protein expression of p38, ABCG2 and NF-kappa B.Significance: Quercetin alone exhibited significant cytotoxic effects on CD44(+)/CD133(+). MK plays an important role in the proliferation of CD44(+)/CD133(+) and CD44(+) cells in particular, and quercetin and MK-silencing therapy may be an important strategy in targeting CSCs that play a role in relapse, migration and drug resistance.