Niclosamide Induces Cell Cycle Arrest in G1 Phase in Head and Neck Squamous Cell Carcinoma Through Let-7d/CDC34 Axis
Niclosamide Induces Cell Cycle Arrest in G1 Phase in Head and Neck Squamous Cell Carcinoma Through Let-7d/CDC34 Axis
复制标题
氯硝柳胺通过 Let-7d/CDC34 轴诱导头颈鳞状细胞癌 G1 期细胞周期停滞
DOI：
10.3389/fphar.2018.01544
复制
发表时间：
2019-01-09
影响因子：
5.6
通讯作者：
Guo, Chuanbin
中科院分区：
文献类型：
--
作者：
Han, Zewen;Li, Qingxiang;Guo, Chuanbin
氯硝柳胺是一种传统的抗绦虫药物，具有很强的抗癌活性。我们之前的研究表明，氯硝柳胺在G1期诱导细胞周期阻滞。然而，潜在的机制仍然未知。本研究探讨了氯硝胺诱导头颈部鳞状细胞癌（HNSCC）细胞系G1阻滞的分子机制。采用诱导细胞ZOOMTM、流式细胞术（FCM）、实时荧光定量PCR （real-time PCR）和western blot方法分析氯硝柳胺对人HNSCC细胞株WSU-HN6和CNE-2Z的影响。荧光素酶测定证实let-7d （let-7家族成员，通过调节细胞周期发挥肿瘤抑制作用）与CDC34 mRNA 3'UTR之间的相互作用。建立异种移植肿瘤模型，评价氯硝柳胺在体内的治疗效果。简而言之，暴露于氯胺酮治疗导致let-7d表达增加，细胞周期调节因子CDC34表达降低，最终导致G1期阻滞。此外，let-7d的过表达诱导G1期阻滞并下调CDC34，而let-7d的下调部分挽救了氯胺酮诱导的G1期阻滞。荧光素酶实验证实通过靶向let-7d直接抑制CDC34。此外，氯硝柳胺通过上调let-7d和下调CDC34显著抑制异种移植物的生长。综上所述，我们的研究结果表明，氯硝柳胺通过let-7d/CDC34轴诱导HNSCC细胞周期阻滞在G1期，这丰富了氯硝柳胺的抗癌机制。
Niclosamide is a traditional anti-tapeworm drug that exhibits potent anti-cancer activity. Our previous study showed that niclosamide induces cell cycle arrest in G1 phase. Nevertheless, the underlying mechanism remains unknown. The following study investigated the molecular mechanism through which niclosamide induced G1 arrest in head and neck squamous cell carcinoma (HNSCC) cell lines. The effect of niclosamide on human HNSCC cell line WSU-HN6 and CNE-2Z were analyzed using IncuCyte ZOOMTM assay, flow cytometry (FCM), real-time PCR and western blot. Luciferase assay was conducted to demonstrate the interaction between let-7d (a let-7 family member which functions as a tumor suppressor by regulating cell cycle) and 3′UTR of CDC34 mRNA. Xenografts tumor model was established to evaluate the niclosamide treatment efficacy in vivo. Briefly, an exposure to niclosamide treatment led to an increased let-7d expression and a decreased expression of cell cycle regulator CDC34, finally leading to G1 phase arrest. Moreover, an overexpression of let-7d induced G1 phase arrest and downregulated CDC34, while the knockdown of let-7d partially rescued the niclosamide-induced G1 phase arrest. Luciferase assay confirmed the direct inhibition of CDC34 through the targeting of let-7d. Furthermore, niclosamide markedly inhibited the xenografts growth through up-regulation of let-7d and down-regulation of CDC34. To sum up, our findings suggest that niclosamide induces cell cycle arrest in G1 phase in HNSCC through let-7d/CDC34 axis, which enriches the anti-cancer mechanism of niclosamide.