长链非编码RNA-NEAT1通过调控miR-124/NF-κB通路促进宫颈癌细胞的迁移和侵袭
Long Non-Coding RNA-NEAT1 Promotes Cell Migration and Invasion via Regulating miR-124/NF-κB Pathway in Cervical Cancer.
作者信息
Shen Xiaofang, Zhao Wei, Zhang Yumei, Liang Bin
机构信息
Department of Obstetrics-Gynecology, Dongying City People's Hospital, Dongying City, Shandong Province 257091, People's Republic of China.
Department of Obstetrics-Gynecology, Dongying City Dongying District People's Hospital, Dongying City, Shandong Province 257000, People's Republic of China.
出版信息
Onco Targets Ther. 2020 Apr 17;13:3265-3276. doi: 10.2147/OTT.S220306. eCollection 2020.
BACKGROUND
This study aimed to investigate the regulatory role of lncRNA-NEAT1 on cervical cancer (CC) and the underlying molecular mechanisms.
METHODS
The expression of lncRNA-NEAT1 and miR-124 was detected in CC tissues and cells (HeLa and SiHa cells) by qRT-RCR. The relation between lncRNA-NEAT1 expression and clinical parameters of CC patients was explored. The cell migration and invasion were detected by wound healing assay and transwell assay. The cell proliferation was detected by CCK-8 and anchorage-independent colony assay. The targeting relation between miR-124 and lncRNA-NEAT1 was predicted by TargetScan and identified by dual luciferase reporter gene and RNA pull-down assay. The expression of metastasis- (MMP-2 and MMP), EMT- (E-cadherin, N-cadherin and Vimentin), and NF-κB pathway-related factors (NF-κB p65, p-NF-κB p65 and IκBα) was detected by Western blot.
RESULTS
The expression of lncRNA-NEAT1 was upregulated in CC tissues and cells and positively correlated with TNM stage and lymph node metastasis. Overexpression of lncRNA-NEAT1 promoted the proliferation, migration and invasion, influenced the expression of EMT markers, and activated NF-κB pathway in HeLa and SiHa cells. Silencing of lncRNA-NEAT1 exhibited opposite effects on HeLa and SiHa cells. LncRNA-NEAT1 could negatively regulate its target miR-124. MiR-124 reversed the effects of lncRNA-NEAT1 on the migration, invasion, EMT and NF-κB pathway of HeLa cells.
CONCLUSION
LncRNA-NEAT1 promoted the migration and invasion of CC cells via regulating miR-124/NF-κB pathway.
背景
本研究旨在探讨长链非编码RNA-NEAT1对宫颈癌（CC）的调控作用及其潜在分子机制。
方法
采用qRT-RCR检测CC组织及细胞（HeLa和SiHa细胞）中lncRNA-NEAT1和miR-124的表达。探讨lncRNA-NEAT1表达与CC患者临床参数的关系。采用伤口愈合试验和Transwell试验检测细胞迁移和侵袭能力。采用CCK-8和非锚定依赖集落试验检测细胞增殖能力。通过TargetScan预测miR-124与lncRNA-NEAT1的靶向关系，并通过双荧光素酶报告基因和RNA下拉试验进行鉴定。采用蛋白质免疫印迹法检测转移相关因子（MMP-2和MMP）、上皮-间质转化相关因子（E-钙黏蛋白、N-钙黏蛋白和波形蛋白）及NF-κB通路相关因子（NF-κB p65、p-NF-κB p65和IκBα）的表达。
结果
lncRNA-NEAT1在CC组织和细胞中表达上调，与TNM分期和淋巴结转移呈正相关。lncRNA-NEAT1过表达促进HeLa和SiHa细胞增殖、迁移和侵袭，影响上皮-间质转化标志物的表达，并激活NF-κB通路。lncRNA-NEAT1沉默对HeLa和SiHa细胞表现出相反的作用。lncRNA-NEAT1可负向调节其靶标miR-124。miR-124可逆转lncRNA-NEAT1对HeLa细胞迁移、侵袭、上皮-间质转化及NF-κB通路的影响。
结论
lncRNA-NEAT1通过调控miR-124/NF-κB通路促进CC细胞的迁移和侵袭。
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