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miR-136-5p/ROCK1轴抑制头颈部癌细胞的侵袭和迁移,并增强其对顺铂的敏感性。

The miR-136-5p/ROCK1 axis suppresses invasion and migration, and enhances cisplatin sensitivity in head and neck cancer cells.

作者信息

Yang Bo, Zang Jian, Yuan Weili, Jiang Xuejun, Zhang Fang

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China.

Department of Oral and Maxillofacial Surgery, The Fourth Affiliated Hospital of China Medical University, Shenyang, Liaoning 110032, P.R. China.

出版信息

Exp Ther Med. 2021 Apr;21(4):317. doi: 10.3892/etm.2021.9748. Epub 2021 Feb 3.

Abstract

Laryngeal squamous cell carcinoma (LSCC) and hypopharyngeal squamous cell carcinoma (HPSCC) are two types of head and neck cancers with high incidence rates and relatively poor prognoses. The aim of the present study was to determine the effects of microRNA (miR/miRNA)-136-5p and its downstream target, Rho-associated coiled-coil containing protein kinase 1 (ROCK1), on LSCC and HPSCC progression and cisplatin sensitivity. The miRNA and protein expression levels in head and neck cancer cell lines were evaluated using reverse transcription-quantitative PCR and western blotting, respectively. MTT, wound healing assays, transwell assays and flow cytometry analysis were performed to measure cell properties. The binding between miR-136-5p and ROCK1 was detected using a dual-luciferase reporter assay. Autophagy double-labeled adenoviral infection assays were used to assess cell autophagy. The results showed that miR-136-5p was expressed in LSCC and HPSCC cells. Functional experiments showed that the expression of miR-136-5p in LSCC and HPSCC cells was negatively correlated with cell viability, invasion and migration. Additionally, miR-136-5p overexpression inhibited epithelial-mesenchymal transition, whereas miR-136-5p knockdown had the opposite effect. Dual-luciferase reporter assays confirmed the targeting relationship between miR-136-5p and ROCK1. miR-136-5p overexpression increased the cisplatin sensitivity of LSCC and HPSCC cells by reducing cell viability, as well as promoting cell apoptosis and autophagy. miR-136-5p overexpression decreased the expression levels of its downstream target ROCK1 and attenuated activity of the Akt/mTOR signaling pathway in cisplatin-treated LSCC and HPSCC cells. Conversely, miR-136-5p knockdown increased ROCK1 levels and decreased cisplatin sensitivity of the LSCC and HPSCC cells by increasing cell viability and inhibiting cell apoptosis, which was reversed by ROCK1 inhibition using the ROCK1 inhibitor, Y27632. Taken together, the results showed that the miR-136-5p/ROCK1 axis inhibits cell invasion and migration, and increases the sensitivity of LSCC and HPSCC cells to cisplatin.

摘要

喉鳞状细胞癌(LSCC)和下咽鳞状细胞癌(HPSCC)是两种发病率较高且预后相对较差的头颈癌。本研究的目的是确定微小RNA(miR/miRNA)-136-5p及其下游靶点含 Rho 相关卷曲螺旋结构蛋白激酶 1(ROCK1)对 LSCC 和 HPSCC 进展及顺铂敏感性的影响。分别使用逆转录定量 PCR 和蛋白质印迹法评估头颈癌细胞系中的 miRNA 和蛋白质表达水平。进行 MTT 法、伤口愈合试验、Transwell 试验和流式细胞术分析以检测细胞特性。使用双荧光素酶报告基因检测法检测 miR-136-5p 与 ROCK1 之间的结合。采用自噬双标记腺病毒感染试验评估细胞自噬。结果显示 miR-136-5p 在 LSCC 和 HPSCC 细胞中表达。功能实验表明,miR-136-5p 在 LSCC 和 HPSCC 细胞中的表达与细胞活力、侵袭和迁移呈负相关。此外,miR-136-5p 过表达抑制上皮-间质转化,而 miR-136-5p 敲低则产生相反的效果。双荧光素酶报告基因检测法证实了 miR-136-5p 与 ROCK1 之间的靶向关系。miR-136-5p 过表达通过降低细胞活力、促进细胞凋亡和自噬,提高了 LSCC 和 HPSCC 细胞对顺铂的敏感性。miR-136-5p 过表达降低了其下游靶点 ROCK1 的表达水平,并减弱了顺铂处理的 LSCC 和 HPSCC 细胞中 Akt/mTOR 信号通路的活性。相反,miR-136-5p 敲低增加了 ROCK1 水平,并通过提高细胞活力和抑制细胞凋亡降低了 LSCC 和 HPSCC 细胞对顺铂的敏感性,而使用 ROCK1 抑制剂 Y27632 抑制 ROCK1 可逆转这种情况。综上所述,结果表明 miR-136-5p/ROCK1 轴抑制细胞侵袭和迁移,并增加 LSCC 和 HPSCC 细胞对顺铂的敏感性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2bd2/7885062/c24e849d6b36/etm-21-04-09748-g00.jpg

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