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MicroRNA-424-5p 通过与 ITGB1 结合抑制非小细胞 LCa 的发展。

MicroRNA-424-5p inhibits the development of non-small cell LCa by binding to ITGB1.

机构信息

Department of Respiratory Medicine, The First People's Hospital of Fuyang, Hangzhou, China.

出版信息

Eur Rev Med Pharmacol Sci. 2019 Oct;23(20):8921-8930. doi: 10.26355/eurrev_201910_19289.

Abstract

OBJECTIVE

The aim of this study was to explore the effect of microRNA-424-5p on the proliferation and apoptosis of non-small cell lung cancer (NSCLC) cells, and to investigate its influence on the expression of ITGB1 and potential regulatory mechanism.

PATIENTS AND METHODS

Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was used to detect the level of microRNA-424-5p in 44 paired NSCLC tissues and adjacent tissues. The relation between microRNA-424-5p expression and NSCLC clinical indicators was analyzed. Subsequently, microRNA-424-5p mimics and inhibitors were transfected into NSCLC cells to construct microRNA-424-5p overexpression or knockdown models, respectively. QRT-PCR was used to further verify the transfection efficiency. A series of experiments, including cell counting kit-8 (CCK-8) assay, colony formation, 5-Ethynyl-2'-deoxyuridine (EdU), and flow cytometry were used to analyze the effect of microRNA-424-5p on the biological function of NSCLC A549 and H358 cells. Finally, the potential association between microRNA-424-5p and its downstream gene ITGB1 was explored through luciferase reporter gene assay and cell recovery experiment.

RESULTS

QRT-PCR results showed that microRNA-424-5p level was significantly lower in NSCLC tissues than that of adjacent normal tissues. Compared with patients with high expression of microRNA-424-5p, the pathological stage of those with low expression of microRNA-424-5p was significantly higher. In vitro experiments showed that microRNA-424-5p overexpression remarkably decreased cell proliferation and increased cell apoptosis, which were further validated in microRNA-424-5p inhibitor group. Subsequently, ITGB1 expression was found significantly up-regulated in NSCLC cell lines and tissues. Meanwhile, ITGB1 expression was negatively correlated with microRNA-424-5p level. In addition, a recovery experiment indicated that overexpression of ITGB1 could counteract the effect of microRNA-424-5p mimics on the proliferation and apoptosis of NSCLC cells. All these findings revealed that microRNA-424-5p and ITGB1 affected the malignant progression of NSCLC.

CONCLUSIONS

MicroRNA-424-5p was closely correlated with the pathological stage and poor prognosis of NSCLC, thereby inhibiting the occurrence and development of NSCLC.

摘要

目的

本研究旨在探讨 microRNA-424-5p 对非小细胞肺癌(NSCLC)细胞增殖和凋亡的影响,并探讨其对 ITGB1 表达的影响及其潜在调控机制。

患者与方法

采用实时荧光定量聚合酶链反应(qRT-PCR)检测 44 对 NSCLC 组织及其相邻组织中 microRNA-424-5p 的水平。分析 microRNA-424-5p 表达与 NSCLC 临床指标的关系。随后,将 microRNA-424-5p 模拟物和抑制剂转染至 NSCLC 细胞中,分别构建 microRNA-424-5p 过表达或敲低模型,进一步采用 qRT-PCR 验证转染效率。采用细胞计数试剂盒-8(CCK-8)检测、集落形成、5-乙炔基-2'-脱氧尿苷(EdU)和流式细胞术分析 microRNA-424-5p 对 NSCLC A549 和 H358 细胞生物学功能的影响。最后,通过荧光素酶报告基因检测和细胞回收实验探讨 microRNA-424-5p 与其下游基因 ITGB1 之间的潜在关联。

结果

qRT-PCR 结果显示,microRNA-424-5p 在 NSCLC 组织中的水平明显低于相邻正常组织。与高表达 microRNA-424-5p 的患者相比,低表达 microRNA-424-5p 的患者的病理分期明显较高。体外实验显示,microRNA-424-5p 过表达显著降低细胞增殖,增加细胞凋亡,这在 microRNA-424-5p 抑制剂组中得到进一步验证。随后,在 NSCLC 细胞系和组织中发现 ITGB1 表达明显上调。同时,ITGB1 表达与 microRNA-424-5p 水平呈负相关。此外,恢复实验表明,ITGB1 的过表达可以抵消 microRNA-424-5p 模拟物对 NSCLC 细胞增殖和凋亡的影响。所有这些发现表明,microRNA-424-5p 和 ITGB1 影响 NSCLC 的恶性进展。

结论

microRNA-424-5p 与 NSCLC 的病理分期和不良预后密切相关,从而抑制 NSCLC 的发生和发展。

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