Wang Jingjie, Zeng Huijuan, Li Hanjun, Chen Tao, Wang Lulu, Zhang Kai, Chen Jing, Wang Rui, Li Qiurong, Wang Shaohua
Department of General Surgery, Jinling Hospital, Medical school of Nanjing University, Nanjing, China.
Department of Medical Oncology, Jinling Hospital, Medical school of Nanjing University, Nanjing, China.
Cell Physiol Biochem. 2017;43(2):717-732. doi: 10.1159/000481445. Epub 2017 Sep 27.
Increasing evidence has demonstrated that microRNAs play a critical role in breast cancer (BC) progression. microRNA-101 (miR-101) has been considered a tumor suppressive miRNA in different cancer types. This study aimed to investigate miR-101 expression in BC tissues and to investigate its roles in BC progression that are mediated by Sex-determining region Y-box 2 (SOX2), a critical oncogene in various cancers.
qRT-PCR and immunohistochemistry were performed to detect miR-101 and SOX2 expression in BC tissues and paired normal tissues or BC cell lines. MTT, transwell migration, wound healing, colony formation, flow cytometric and xenograft assays were performed to determine the influence of miR-101 and SOX2 on the malignant behaviors of BC cells in vitro and in vivo.
miR-101 was significantly downregulated in BC tissues and cell lines. miR-101 overexpression inhibited the malignant behaviors of BC cells, both in vitro and in vivo. miR-101 downregulation had the converse effect. A miR-101 binding site was identified by luciferase reporter assay in the 3'UTR of SOX2. SOX2 was upregulated in BC tissues and cell lines, and its upregulation was associated with lymph node metastasis, pathological grade and TNM classification. SOX2 knockdown mimicked the effects of miR-101 overexpression on the malignant behaviors of BC cells, while SOX2 overexpression mitigated the miR-101-induced inhibition of these effects.
Our study revealed that miR-101 plays a critical role in suppressing tumor progression by directly targeting SOX2.
越来越多的证据表明,微小RNA在乳腺癌(BC)进展中起关键作用。微小RNA-101(miR-101)在不同癌症类型中被认为是一种肿瘤抑制性微小RNA。本研究旨在调查miR-101在BC组织中的表达,并研究其在BC进展中由性别决定区Y盒2(SOX2)介导的作用,SOX2是多种癌症中的关键致癌基因。
采用qRT-PCR和免疫组织化学检测BC组织及配对的正常组织或BC细胞系中miR-101和SOX2的表达。进行MTT、Transwell迁移、伤口愈合、集落形成、流式细胞术和异种移植试验,以确定miR-101和SOX2对BC细胞体外和体内恶性行为的影响。
miR-101在BC组织和细胞系中显著下调。miR-101过表达在体外和体内均抑制了BC细胞的恶性行为。miR-101下调则产生相反的效果。通过荧光素酶报告基因检测在SOX2的3'UTR中鉴定出一个miR-101结合位点。SOX2在BC组织和细胞系中上调,其上调与淋巴结转移、病理分级和TNM分类相关。SOX2敲低模拟了miR-101过表达对BC细胞恶性行为的影响,而SOX2过表达减轻了miR-101诱导的对这些影响的抑制作用。
我们的研究表明,miR-101通过直接靶向SOX2在抑制肿瘤进展中起关键作用。