circFOXP1靶向miR-4429调控鼻咽癌6-10B细胞的增殖、迁移及侵袭^*

circFOXP1 Targets miR-4429 to Regulate Proliferation, Migration and Invasion of Nasopharyngeal Carcinoma 6-10B Cells

摘要

目的 探讨环状RNA叉头框蛋白P1（circular RNA forkhead box protein P1, circFOXP1）是否通过靶向miR-4429调控鼻咽癌6-10B细胞的增殖、迁移及侵袭。方法 分析circFOXP1和miR-4429表达与鼻咽癌患者病理特征的关系。在鼻咽癌6-10B细胞中分别转染si-NC、si-circFOXP1、pcDNA、pcDNA-circFOXP1、miR-NC、miR-4429模拟物，或同时转染si-circFOXP1与anti-miR-NC、anti-miR-4429。应用实时定量聚合酶链反应（qRT-PCR）检测鼻咽癌组织中circFOXP1和miR-4429的表达水平；采用细胞计数试剂盒8（CCK-8）、克隆形成实验、划痕实验及Transwell实验分别评估细胞活力、克隆形成能力、迁移能力及侵袭能力；Western blot法检测上皮型钙黏蛋白（E-cadherin）和神经型钙黏蛋白（N-cadherin）的表达；双萤光素酶报告基因实验验证circFOXP1与miR-4429的靶向关系。结果 鼻咽癌组织中circFOXP1的表达水平（4.61±0.31）较癌旁组织（1.00±0.05）升高（P<0.001），miR-4429的表达水平（0.37±0.03）较癌旁组织（1.00±0.08）降低（P<0.001）。circFOXP1表达与TNM分期相关（P<0.05），与肿瘤体积无显著相关性（P>0.05）；miR-4429表达与肿瘤体积和TNM分期均相关（P<0.05）。抑制circFOXP1表达或过表达miR-4429均可降低鼻咽癌6-10B细胞的活力、克隆形成数、划痕愈合率、侵袭细胞数及N-cadherin蛋白表达，同时升高E-cadherin蛋白表达（P<0.05）。circFOXP1可靶向并负向调控miR-4429的表达。干扰miR-4429能够逆转circFOXP1沉默对鼻咽癌细胞增殖、迁移及侵袭的抑制作用（P<0.05）。结论 抑制circFOXP1表达可通过靶向miR-4429减弱鼻咽癌6-10B细胞的增殖、迁移及侵袭能力。

Abstract

Objective To determine whether circular RNA forkhead box protein P1 (circFOXP1) regulates the proliferation, migration and invasion of nasopharyngeal carcinoma 6-10B cells by targeting miR-4429. Methods The correlations of circFOXP1 and miR-4429 with pathological characteristics of nasopharyngeal carcinoma patients were analyzed. Nasopharyngeal carcinoma 6-10B cells were transfected with si-NC, si-circFOXP1, pcDNA, pcDNA-circFOXP1, miR-NC, miR-4429 mimics, or co-transfected with si-circFOXP1 and anti-miR-NC, or si-circFOXP1 and anti-miR-4429. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect circFOXP1 and miR-4429 expression in nasopharyngeal carcinoma tissues. Cell viability, colony formation, migration and invasion abilities were assessed by Cell Counting Kit-8 (CCK-8), colony formation, wound healing and Transwell assays. Western blotting was used to detect E-cadherin and N-cadherin protein expression. A dual-luciferase reporter assay verified the targeting relationship betweencircFOXP1 and miR-4429. Results The expression level of circFOXP1 in nasopharyngeal carcinoma tissues (4.61 ± 0.31) was significantly higher than that in adjacent tissues (1.00 ± 0.05) (P < 0.001), while the expression level of miR-4429 (0.37 ± 0.03) was significantly lower than that in adjacent tissues (1.00 ± 0.08) ( P < 0.001). The expression of miR-4429 was correlated with tumor volume and TNM stage ( P < 0.05), while the expression of circFOXP1 was correlated with TNM stage(P < 0.05) but not with tumor volume ( P > 0.05). Inhibition of circFOXP1 expression or overexpression of miR-4429 reduced cell viability, colony formation number, wound healing rate, invasive cell number and N-cadherin protein expression, while increased E-cadherin protein expression in nasopharyngeal carcinoma 6-10B cells (P < 0.05). circFOXP1 targeted and negatively regulated miR-4429 expression. Interference with miR-4429 reversed the inhibitory effects of circFOXP1 silencing on proliferation, migration and invasion of nasopharyngeal carcinoma cells (P < 0.05). Conclusion Inhibition of circFOXP1 expression can attenuate the proliferation, migration and invasion of nasopharyngeal carcinoma 6-10B cells by targeting miR-4429.

关键词

鼻咽癌6-10B细胞 / circFOXP1 / miR-4429 / 增殖 / 迁移 / 侵袭

Key words

Nasopharyngeal carcinoma 6-10B cells / circFOXP1 / miR-4429 / Proliferation / Migration / Invasion

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基金资助

*河南省医学科技攻关计划项目(LHGJ20210911)