Molecular mechanisms of long non-coding RNA ZEB1-antisense RNA1 in regulating lymphoma cell proliferation, invasion, and migration by targeting microRNA-224
- VernacularTitle:长链非编码RNA ZEB1反义RNA1通过靶向微小RNA-224调控淋巴瘤细胞增殖、侵袭、迁移的分子机制研究
- Author:
Dan SHI
1
;
Chao SHEN
Author Information
- Publication Type:Research Article
- Keywords: long non-coding RNA ZEB1-antisense RNA1; lymphoma; microRNA-224; proliferation; migration; invasion
- From: Journal of Clinical Medicine in Practice 2024;28(22):35-40
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the molecular mechanisms of long non-coding RNA ZEB1-antisense RNA1 (LncRNA ZEB1-AS1) in regulating lymphoma cell proliferation, invasion, and migration by targeting microRNA (miR)-224. Methods The lymphoma cell line Raji was cultured
in vitro and transfected with si-NC, si-ZEB1-AS1, miR-NC, miR-224 mimics, si-ZEB1-AS1+anti-miR-NC, and si-ZEB1-AS1+anti-miR-224, respectively. Real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) was used to assess the expression levels of LncRNA ZEB1-AS1 and miR-224 in lymphoma cells. Cell viability was detected using the Cell Counting Kit-8 (CCK-8). Cell migration and invasion abilities were evaluated by Transwell experiments. Dual luciferase reporter experiments were conducted to confirm the regulatory effect of LncRNA ZEB1-AS1 on miR-224. Western blot was performed to detect the expression levels of cyclin D1 (CyclinD1), matrix metalloproteinase (MMP)-2, and MMP-9 proteins. Results Compared with the control group, the lymphoma group showed increased LncRNA ZEB1-AS1 levels and decreased miR-224 expression levels (P < 0.05). Compared with the si-NC group, the si-ZEB1-AS1 group exhibited decreased expression levels of LncRNA ZEB1-AS1, optical density (OD) values, the number of migrated and invaded cells, and expression levels of CyclinD1, MMP-2, and MMP-9 protein (P < 0.05). Compared with the miR-NC group, the miR-224 group showed increased miR-224 expression levels but decreased OD values, the number of migrated and invaded cells, and expression levels of CyclinD1, MMP-2, and MMP-9 proteins (P < 0.05). Upregulation of miR-224 levels reduced the luciferase activity of WT-ZEB1-AS1 but had no effect on MUT-ZEB1-AS1 luciferase activity. Compared with the pcDNA-NC group, the pcDNA-ZEB1-AS1 group showed decreased miR-224 expression levels (P < 0.05); compared with the si-NC group, the si-ZEB1-AS1 group showed increased miR-224 expression levels (P < 0.05). Compared with the si-ZEB1-AS1+anti-miR-NC group, the si-ZEB1-AS1+anti-miR-224 group showed increased OD values, the number of migrated and invaded cells, and expression levels of CyclinD1, MMP-2, and MMP-9 proteins(P < 0.05). Conclusion Inhibition of ZEB1-AS1 to targetedly upregulate of miR-224 expression can effectively reduce lymphoma cell proliferation, invasion, and migration abilities.
