Abstract:

Objective　To investigate the effect of long non-coding RNA (lncRNA) insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) -AS1 on the migration and proliferation of laryngeal cancer cells by regulating the expression of microRNA-375 (miR-375). Methods　The research period was from January 2022 to November 2023, the research place was the central laboratory of Luoyang Central Hospital Affiliated to Zhengzhou University, and the research type was basic research. qPCR was used to detect the expression of IGF2BP2-AS1 in laryngeal cancer cell lines AMC-NH-8, TU159, TU138, and TU686 and bronchial epithelial cell line 16HBE. Taking TU686 cells as the research object, the TU686 cells were divided into a si-NC group and a si-IGF2BP2-AS1 group, and IGF2BP2-AS1 small interfering RNA was used to down-regulate the expression level of IGF2BP2-AS1. The scratch healing assay and CCK-8 method were used to analyze the effects of knocking down IGF2BP2-AS1 on the migration and proliferation of TU686 cells. Dual luciferase reporter gene experiment verified the targeting relationship between IGF2BP2-AS1 and miR-375. qPCR was used to detect the effect of knocking down IGF2BP2-AS1 on the expression of miR-375. Western blotting was used to detect the expressions of migration proteins (N-Cadherin and FOXC2) and proliferation proteins (CDK2 and Cyclin A) in cells. One-way ANOVA and t test were used. Results　Compared with 16HBE cells, IGF2BP2-AS1 was highly expressed in laryngeal cancer cell lines (F=37.42, P<0.01). The scratch healing rate of TU686 cells in the si-IGF2BP2-AS1 group was lower than that in the si-NC group [(28.16±6.13)% vs. (65.08±3.82)%], with a statistically significant difference (t=5.11, P<0.01). The proliferation capacity of TU686 cells in the si-IGF2BP2-AS1 group was lower than that in the si-NC group 2, 3, 4, and 5 days after planking (t=3.83, 3.17, 3.02, and 6.31, all P<0.01). In IGF2BP2-AS1-Wt, the relative luciferase activity of the miR-375 group was lower than that of the miR-NC group (t=8.95, P<0.01). The expression of miR-375 in TU686 cells in the si-IGF2BP2-AS1 group was higher than that in the si-NC group [(4.95±0.49) vs. (1.02±0.40)], with a statistically significant difference (t=6.23, P<0.01). Compared with the si-NC group, the expression levels of migration proteins (N-Cadherin and FOXC2) and proliferation proteins (CDK2 and Cyclin A) were reduced in TU686 cells after knocking down IGF2BP2-AS1. Conclusion　IGF2BP2-AS1 is highly expressed in laryngeal cancer cell lines, and knocking down IGF2BP2-AS1 inhibits the migration and proliferation of laryngeal cancer TU686 cells by regulating the expression of miR-375.

Key words:

Laryngeal cancer, Long-chain non-coding RNA, IGF2BP2-AS1, miR-375, Migration, Proliferation

摘要：

目的　探讨长链非编码RNA（lncRNA）胰岛素样生长因子2 mRNA结合蛋白2（IGF2BP2）-AS1通过调控微RNA-375（miR-375）表达对喉癌细胞迁移和增殖的影响。方法　研究时间为2022年1月至2023年11月，研究地点为郑州大学附属洛阳中心医院中心实验室，研究类型为基础研究。实时荧光定量PCR（qPCR）检测喉癌细胞系AMC-NH-8、TU159、TU138、TU686和支气管上皮细胞系16HBE中IGF2BP2-AS1的表达。以TU686细胞为研究对象，将TU686细胞分为si-NC组和si-IGF2BP2-AS1组，采用IGF2BP2-AS1小干扰RNA下调IGF2BP2-AS1表达水平。划痕愈合实验和CCK-8法分析敲降IGF2BP2-AS1对TU686细胞迁移及增殖的影响。双荧光素酶报告基因实验验证IGF2BP2-AS1与miR-375的靶向关系。qPCR检测敲降IGF2BP2-AS1对miR-375表达的影响。Western blotting检测细胞中迁移蛋白N-钙黏着蛋白（N-Cadherin）、叉头框蛋白C2（FOXC2）和增殖蛋白CDK2、细胞周期蛋白A（Cyclin A）的表达。统计学方法采用单因素方差分析、t检验。结果　与16HBE细胞相比，IGF2BP2-AS1在喉癌细胞系中均高表达（F=37.42，P<0.01）。si-IGF2BP2-AS1组TU686细胞划痕愈合率低于si-NC组［（28.166.13）%比（65.083.82）%］，差异有统计学意义（t=5.11，P<0.01）。在铺板后第2、3、4、5天，si-IGF2BP2-AS1组TU686细胞增殖能力均低于si-NC组（t=3.83、3.17、3.02、6.31，均P<0.01）。IGF2BP2-AS1-Wt中，miR-375组相对荧光素酶活性低于miR-NC组（t=8.95，P<0.01）。si-IGF2BP2-AS1组TU686细胞中miR-375表达高于si-NC组［（4.950.49）比（1.020.40）］，差异有统计学意义（t=6.23，P<0.01）。敲降IGF2BP2-AS1后TU686细胞中迁移蛋白N-Cadherin、FOXC2表达与增殖蛋白CDK2、Cyclin A表达均低于si-NC组。结论　IGF2BP2-AS1在喉癌细胞系中呈高表达，敲降IGF2BP2-AS1通过调控miR-375表达抑制喉癌TU686细胞的迁移及增殖能力。

关键词:

喉癌, 长链非编码RNA, IGF2BP2-AS1, miR-375, 迁移, 增殖