Inhibitory action of pristimerin on hypoxia-mediated metastasis involves stem cell characteristics and EMT in PC-3 prostate cancer cells
Inhibitory action of pristimerin on hypoxia-mediated metastasis involves stem cell characteristics and EMT in PC-3 prostate cancer cells
复制标题
pritimerin 对缺氧介导的转移的抑制作用涉及 PC-3 前列腺癌细胞中的干细胞特征和 EMT
DOI：
10.3892/or.2015.3708
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发表时间：
2015-03-01
期刊：
影响因子：
4.2
通讯作者：
Xu, Dongliang
中科院分区：
文献类型：
--
作者：
Zuo, Jianwei;Guo, Yuanqing;Xu, Dongliang
本研究的目的是探讨是否pristimerin影响骨转移，干细胞特性和上皮间质转化（EMT）的前列腺癌（PCa）PC-3细胞缺氧。将PC-3细胞在缺氧或常氧条件下培养48小时，然后在常氧条件下用浓度从0增加到0.8 μ mol/l的pristimerin处理。Western blotting检测缺氧诱导因子-1 α（HIF-1 α）表达。用CCK-8测定法评估增殖。Transwell侵袭实验检测细胞的侵袭能力。通过球体形成、集落形成试验和蛋白质印迹法检测干细胞特征，包括干细胞特征相关标志物CD 44、KLF 4、OCT 4和AG 02。EMT相关标志物包括N-cadherin、fibronectin、vimentin和ZEB 1的表达变化通过Western blotting检测证实。在培养液中加入pristimerin可剂量依赖性地抑制缺氧诱导的PC-3细胞增殖。Pristimerin能有效抑制PCa细胞在体外缺氧诱导的侵袭。此外，用pristimerin处理细胞诱导缺氧诱导的干细胞特性和EMT的逆转，这通过球体形成、集落形成测定以及CSC和EMT相关标志物的表达变化来证实。低剂量pristimerin逆转PCa细胞缺氧诱导的干细胞特性和EMT具有剂量依赖性。这些结果表明，用pristimerin治疗可能是通过逆转缺氧诱导的干细胞特征和癌细胞中的EMT来抑制缺氧诱导的转移的潜在策略，这证明了pristimerin作为PCa患者的实际化学预防方法的潜在用途。
The aim of the present study was to investigate whether pristimerin affects the bone metastasis, stem cell characteristics and epithelial-mesenchymal transition (EMT) of prostate cancer (PCa) PC-3 cells subjected to hypoxia. The PC-3 cells were cultured under hypoxia or normoxia for 48 h and were then treated with increasing concentrations of pristimerin from 0 to 0.8 mu mol/l, under normoxia. Hypoxia-inducible factor-1 alpha (HIF-1 alpha) was detected by western blotting. Proliferation was assessed with the CCK-8 assay. Transwell invasion assay was used to analyze the potency of invasion. Stem cell characteristics were detected by sphere formation, colony formation assay and western blotting, including CD44, KLF4, OCT4 and AG02, which are stem cell characteristic-related markers. EMT was confirmed by the expression changes of EMT-related markers, including N-cadherin, fibronectin, vimentin and ZEB1, which were evaluated by western blotting. The addition of pristimerin to the medium reduced the hypoxia-induced PC-3 cell proliferation in a dose-dependent manner. Pristimerin effectively inhibited hypoxia-induced invasion of the PCa cells in vitro. Moreover, the treatment of cells with pristimerin induced the reversal of hypoxia-induced stem cell characteristics and EMT, which was confirmed by sphere formation, colony formation assay and the expression changes of CSC- and EMT-related markers. The reversal of hypoxia-induced stem cell characteristics and EMT in the PCa cells by low-dose pristimerin was dose-dependent. These results showed that treatment with pristimerin may be a potential strategy for the suppression of hypoxia-induced metastasis through the reversal of hypoxia-induced stem cell characteristics and EMT in cancer cells, which justifies the potential use of pristimerin as a practical chemopreventive approach for patients with PCa.