Peng Qian, Zhang Liyang, Li Junjun, Wang Wei, Cai Jing, Ban Yuanyuan, Zhou Ying, Hu Meng, Mei Yan, Zeng Zhaoyang, Li Xiaoling, Xiong Wei, Li Guiyuan, Tan Yixin, Xiang Bo, Yi Mei
Hunan Provincial Cancer Hospital and Cancer Hospital Affiliated to Xiangya Medical School, Central South University, Changsha 410013, Hunan, China.
The Key Laboratory of Carcinogenesis and Cancer Invasion of the Chinese Ministry of Education, Cancer Research Institute and School of Basic Medical Sciences, Central South University, Changsha, 410078, Hunan, China.
J Cancer. 2020 Feb 10;11(9):2485-2495. doi: 10.7150/jca.40709. eCollection 2020.
: Nasopharyngeal carcinoma (NPC) is a unique subtype of head and neck cancer, within highest incidence in South China and southeastern Asia but rare in other regions worldwide. FOXA1 is a pioneer factor implicated in various human malignancies. Downregulation of FOXA1 promotes NPC cells proliferation, invasiveness in vitro and tumorigenicity in vivo. However, it is remain elusive to determine whether microRNAs (miRNAs) regulated by FOXA1 contribute to NPC progression. : In this study, differentially expressed miRNAs and mRNAs induced by FOXA1 expression were determined by microarray. Integrative miRNA-mRNA regulatory networks mediated by FOXA1 in NPC were established. The expressions of differentially expressed miRNAs in NPC cells were measured by quantitative reverse-transcription PCR. Cell viability was determined by CCK-8 assays. Cell migration and invasiveness were measured by Transwell assays. The correlation between miRNAs and its target mRNAs was analyzed. : FOXA1 suppressed the expression of miR-100-5p and miR-125b-5p in NPC cells. Silencing either miR-100-5p or miR-125b-5p inhibited the malignant behaviors of NPC cells, whereas re-expression of miR-100-5p or miR-125b-5p restored the malignancy of NPC cells repressed by FOXA1. Mechanistically, miR-100-5p or miR-125b-5p suppressed RASGRP3 or FOXN3 expression respectively via direct binding to its 3'-UTR. Furthermore, we demonstrated that FOXA1 induced RASGRP3 or FOXN3 expression via inhibiting miR-100-5p or miR-125b-5p. Upregulation of RASGRP3 or FOXN3 contributed to inhibition of NPC by FOXA1. We also demonstrated that the mRNA levels of RASGRP3 and FOXN3 are positively correlated with FOXA1. : Our study provided evidence the first time that FOXA1 suppresses NPC cells via downregulation of miR-100-5p or miR-125b-5p.
鼻咽癌（NPC）是头颈癌的一种独特亚型，在中国南方和东南亚发病率最高，而在世界其他地区则较为罕见。FOXA1是一种与多种人类恶性肿瘤相关的先驱因子。FOXA1的下调促进了NPC细胞的增殖、体外侵袭性和体内致瘤性。然而，由FOXA1调控的微小RNA（miRNA）是否有助于NPC进展仍不清楚。
在本研究中，通过微阵列确定了由FOXA1表达诱导的差异表达miRNA和mRNA。建立了FOXA1在NPC中介导的综合miRNA-mRNA调控网络。通过定量逆转录PCR测量NPC细胞中差异表达miRNA的表达。通过CCK-8测定法测定细胞活力。通过Transwell测定法测量细胞迁移和侵袭性。分析了miRNA与其靶mRNA之间的相关性。
FOXA1抑制NPC细胞中miR-100-5p和miR-125b-5p的表达。沉默miR-100-5p或miR-125b-5p均可抑制NPC细胞的恶性行为，而重新表达miR-100-5p或miR-125b-5p可恢复被FOXA1抑制的NPC细胞的恶性程度。机制上，miR-100-5p或miR-125b-5p分别通过直接结合其3'-UTR抑制RASGRP3或FOXN3的表达。此外，我们证明FOXA1通过抑制miR-100-5p或miR-125b-5p诱导RASGRP3或FOXN3的表达。RASGRP3或FOXN3的上调有助于FOXA1对NPC的抑制作用。我们还证明RASGRP3和FOXN3的mRNA水平与FOXA1呈正相关。
我们的研究首次提供证据表明FOXA1通过下调miR-100-5p或miR-125b-5p抑制NPC细胞。
