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微小RNA-150靶向Rho相关蛋白激酶1以抑制甲状腺乳头状癌的细胞增殖、迁移和侵袭。

MicroRNA-150 targets Rho-associated protein kinase 1 to inhibit cell proliferation, migration and invasion in papillary thyroid carcinoma.

作者信息

Cheng Lixia, Zhou Ruixiu, Chen Min, Feng Linan, Li Hongyan

机构信息

Department of Endocrine, Weifang People's Hospital, Weifang Medical University, Weifang, Shandong 261000, P.R. China.

Department of Endocrine, Gaomi People's Hospital, Weifang Medical University, Weifang, Shandong 261000, P.R. China.

出版信息

Mol Med Rep. 2017 Aug;16(2):2217-2224. doi: 10.3892/mmr.2017.6842. Epub 2017 Jun 22.

Abstract

Thyroid cancer is the most prevalent malignant tumor of the endocrine organs and accounts for one third of all head and neck tumors. Dysregulation of microRNAs is well‑known to contribute to the development of various cancers, including papillary thyroid carcinoma (PTC), which accounts for 80‑90% of all thyroid cancer cases. The present study aimed to investigate the expression, functional roles of microRNA‑150 (miR‑150) and its direct target gene in PTC. miR‑150 expression in PTC tissues and cell lines was analyzed by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). After transfection with miR‑150 mimics, cell proliferation, migration and invasion was analyzed by MTT and Transwell assays, respectively. Bioinformatics analysis was performed to investigate the potential target genes of miR‑150, which were then confirmed by luciferase reporter assay, RT‑qPCR and western blotting. Functional assays were also applied to investigate the effects of endogenous Rho‑associated protein kinase 1 (ROCK1) in PTC. miR‑150 was demonstrated to be significantly downregulated in PTC tissues and cell lines. In addition, reduced miR‑150 expression was obviously correlated with TNM stage and lymph node metastasis in PTC patients. Restoration of miR‑150 expression significantly inhibited PTC cell proliferation, migration and invasion in vitro. Furthermore, ROCK1 was identified as a direct target gene of miR‑150. Therefore, ROCK1 knockdown may serve tumor suppressive functions in PTC, induced by miR‑150 overexpression. In conclusion, miR‑150 overexpression in PTC may inhibit growth and metastasis of PTC cells. miR‑150/ROCK1‑based targeted therapy may be a potential strategy for the treatment of PTC.

摘要

甲状腺癌是内分泌器官中最常见的恶性肿瘤,占所有头颈肿瘤的三分之一。众所周知,微小RNA的失调会促进包括甲状腺乳头状癌(PTC)在内的各种癌症的发展,PTC占所有甲状腺癌病例的80%-90%。本研究旨在探讨微小RNA-150(miR-150)及其直接靶基因在PTC中的表达及功能作用。采用逆转录-定量聚合酶链反应(RT-qPCR)分析PTC组织和细胞系中miR-150的表达。用miR-150模拟物转染后,分别通过MTT和Transwell实验分析细胞增殖、迁移和侵袭情况。进行生物信息学分析以研究miR-150的潜在靶基因,然后通过荧光素酶报告基因实验、RT-qPCR和蛋白质免疫印迹法进行验证。还应用功能实验研究内源性Rho相关蛋白激酶1(ROCK1)在PTC中的作用。结果表明,miR-150在PTC组织和细胞系中显著下调。此外,PTC患者中miR-150表达降低明显与TNM分期和淋巴结转移相关。miR-150表达的恢复显著抑制了PTC细胞在体外的增殖、迁移和侵袭。此外,ROCK1被鉴定为miR-150的直接靶基因。因此,敲低ROCK1可能在miR-150过表达诱导的PTC中发挥肿瘤抑制作用。总之,PTC中miR-150过表达可能抑制PTC细胞的生长和转移。基于miR-150/ROCK1的靶向治疗可能是治疗PTC的潜在策略。

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