miR-501 在宫颈癌中上调，并通过靶向 CYLD 促进细胞增殖、迁移和侵袭。

miR-501 is upregulated in cervical cancer and promotes cell proliferation, migration and invasion by targeting CYLD.

机构信息

Department of Pathology and Forensics, Dalian Medical University, Dalian 116044, China.

Department of Anesthesia, First Affiliated Hospital of Dalian Medical University, Dalian 116011, China.

出版信息

Chem Biol Interact. 2018 Apr 1;285:85-95. doi: 10.1016/j.cbi.2018.02.024. Epub 2018 Feb 23.

DOI:10.1016/j.cbi.2018.02.024

PMID:29477382

Abstract

BACKGROUND

Cervical cancer is the common gynecological deadly malignancy worldwide. Here we attempted to evaluate the effects and mechanisms of microRNA-501-5p (miR-501) on the cell proliferation, migration, invasion and the clinical significance in the cervical cancer.

METHODS

Cervical cancer HeLa cells were transfected with miR-501 mimic or inhibitor or siRNA against Cylindromatosis (CYLD) using Lipofectamine 2000. miR-501 expression was assessed in HeLa cells and cervical cancer specimens by real-time qRT-PCR. The functional roles of miR-501 were determined by CCK-8, colony formation, scratch wound healing and transwell assays. The apoptosis rate was detected by flow cytometry assay. CYLD, BCL-2, BAX, NF-κB p65 and phosphorylated p65 (p-p65) proteins were examined by Western blotting. CYLD expression was evaluated by immunohistochemistry in cervical cancer tissues.

RESULTS

miR-501 was upregulated, whereas CYLD protein was downregulated in cervical cancer tissues compared to normal cervical tissues. miR-501 overexpression and CYLD protein downregulation were positively correlated with poor differentiation, tumor size, International Federation of Gynecology and Obstetrics (FIGO) stage and lymph node metastasis. CYLD was downregulated by miR-501 at both mRNA and protein levels in HeLa cells. miR-501 promoted cell proliferation, migration and invasion in cervical cancer, while inhibited the apoptosis. This is possibly due to the downregulation of CYLD and subsequent activation of NF-κB p65.

CONCLUSIONS

miR-501 upregulation and CYLD downregulation are associated with the development and progression of cervical cancer. miR-501 promotes cervical cancer cell proliferation, migration and invasion possibly via downregulating CYLD and subsequently activating NF-κB p65. miR-501 might be a potential therapeutic target for cervical cancer.

摘要

背景

宫颈癌是全球常见的妇科致命恶性肿瘤。在这里，我们试图评估 microRNA-501-5p（miR-501）对宫颈癌细胞增殖、迁移、侵袭的影响及其临床意义。

方法

用 Lipofectamine 2000 将 miR-501 模拟物或抑制剂或针对 Cylindromatosis（CYLD）的 siRNA 转染入 HeLa 细胞。通过实时 qRT-PCR 评估 HeLa 细胞和宫颈癌标本中的 miR-501 表达。通过 CCK-8、集落形成、划痕愈合和 Transwell 分析测定 miR-501 的功能作用。通过流式细胞术检测细胞凋亡率。通过 Western blot 检测 CYLD、BCL-2、BAX、NF-κB p65 和磷酸化 p65（p-p65）蛋白。通过免疫组织化学法评估宫颈癌组织中 CYLD 的表达。

结果

与正常宫颈组织相比，宫颈癌组织中 miR-501 上调，而 CYLD 蛋白下调。miR-501 过表达和 CYLD 蛋白下调与低分化、肿瘤大小、国际妇产科联合会（FIGO）分期和淋巴结转移呈正相关。miR-501 在 HeLa 细胞中下调 CYLD 的 mRNA 和蛋白水平。miR-501 促进宫颈癌细胞增殖、迁移和侵袭，同时抑制细胞凋亡。这可能是由于 CYLD 的下调和随后的 NF-κB p65 激活。