| 吉 鸿,索 丹,赵 耀,李顺乐,徐 心,常 帅.lncRNA CEBPA-AS1靶向miR-455-3p调控胃癌细胞生物学行为的分子机制研究[J].,2021,(24):4609-4616 |
| lncRNA CEBPA-AS1靶向miR-455-3p调控胃癌细胞生物学行为的分子机制研究 |
| Study on the Molecular Mechanism of lncRNA CEBPA-AS1 on Regulating the Biological Behavior of Gastric Cancer Cells by Targeting miR-455-3p |
| 投稿时间：2021-05-10 修订日期：2021-05-31 |
| DOI：10.13241/j.cnki.pmb.2021.24.002 |
| 中文关键词: lncRNA CEBPA-AS1 miR-455-3p 胃癌 细胞增殖 迁移 侵袭 |
| 英文关键词: lncRNA CEBPA-AS1 miR-455-3p Gastric cancer Cell proliferation Migration Invasion |
| 基金项目:陕西省自然科学基础研究计划项目（2019JQ-967） |
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| 中文摘要: |
| 摘要 目的：探讨lncRNA CEBPA-AS1对胃癌细胞生物学行为的影响及其可能作用机制。方法：qRT-PCR法检测胃癌组织、癌旁组织与正常人胃上皮GES1细胞和人胃癌SNU-1、AGS、HS-746T细胞系中lncRNA CEBPA-AS1、miR-455-3p的表达量。si-NC、si-lncRNA CEBPA-AS1、miR-NC、miR-455-3p mimics、si-lncRNA CEBPA-AS1与anti-miR-NC、si-lncRNA CEBPA-AS1与anti-miR-455-3p分别转染至SNU-1细胞（分别命名为si-NC组、si-lncRNA CEBPA-AS1组、miR-NC组、miR-455-3p组、si-lncRNA CEBPA-AS1+anti-miR-NC组和si-lncRNA CEBPA-AS1+anti-miR-455-3p组）后，MTT实验与平板克隆形成实验分别检测细胞增殖及克隆形成能力，Transwell小室实验检测细胞迁移及侵袭能力，双荧光素酶报告基因实验与qRT-PCR实验验证lncRNA CEBPA-AS1与miR-455-3p的靶向调控关系，Western blot法检测MMP2、MMP9蛋白表达情况。结果：与癌旁组织比较，胃癌组织中lncRNA CEBPA-AS1的表达量显著升高，miR-455-3p的表达量显著降低，差异均有统计学意义（均P＜0.05）。与GES1细胞比较，SNU-1、AGS、HS-746T细胞中lncRNA CEBPA-AS1的表达量显著升高，miR-455-3p的表达量显著降低，其中SNU-1细胞的lncRNA CEBPA-AS1表达量最高，差异均有统计学意义（均P＜0.05）。与si-NC组比较，si-lncRNA CEBPA-AS1组细胞活力降低，细胞克隆形成数、迁移及侵袭细胞数减少，MMP2、MMP9蛋白表达水平降低，差异均有统计学意义（P＜0.05）。与miR-NC组比较，miR-455-3p组细胞活力降低，细胞克隆形成数、迁移及侵袭细胞数减少，MMP2、MMP9蛋白表达水平降低，差异均有统计学意义（P＜0.05）。lncRNA CEBPA-AS1可靶向结合miR-455-3p，并可负调控miR-455-3p的表达。与si-lncRNA CEBPA-AS1+anti-miR-NC组比较，si-lncRNA CEBPA-AS1+anti-miR-455-3p组细胞活力升高，细胞克隆形成数、迁移及侵袭细胞数增多，MMP2、MMP9蛋白表达水平升高，差异均有统计学意义（P＜0.05）。结论：干扰lncRNA CEBPA-AS1表达可通过靶向调控miR-455-3p而抑制胃癌细胞增殖、克隆形成、迁移及侵袭。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the effect of lncRNA CEBPA-AS1 on the biological behavior of gastric cancer cells and its possible mechanism. Methods: qRT-PCR was used to detect the expression levels of lncRNA CEBPA-AS1 and miR-455-3p in gastric cancer tissues, adjacent tissues, normal human gastric epithelial GES1 cells and human gastric cancer cell lines SNU-1, AGS and HS-746T. After si-NC, si-lncRNA CEBPA-AS1, miR-NC, miR-455-3p mimics, si-lncRNA CEBPA-AS1 and anti-miR-NC, si-lncRNA CEBPA-AS1 and anti-miR-455-3p were respectively transfected into SNU-1 cells (which were respectively named as si-NC group, si-lncRNA CEBPA-AS1 group, miR-NC group, miR-455-3p group, si-lncRNA CEBPA-AS1+anti-miR-NC group and si-lncRNA CEBPA-AS1+anti- miR-455-3p group), the MTT experiment and the plate clone formation experiment were used to detect cell proliferation and clone formation ability, the Transwell chamber experiment was used to detect cell migration and invasion ability, the dual-luciferase reporter assay and qRT-PCR experiment were used to verify the targeted regulation relationship between lncRNA CEBPA-AS1 and miR-455-3p, the Western blot method was used to detect the protein expression of MMP2 and MMP9. Results: Compared with adjacent tissues, the expression of lncRNA CEBPA-AS1 in gastric cancer tissues was significantly increased, while the expression of miR-455-3p was significantly decreased, and the differences were statistically significant(P<0.05). Compared with GES1 cells, the expression of lncRNA CEBPA-AS1 in SNU-1, AGS, HS-746T cells was significantly increased, the expression of miR-455-3p was significantly decreased, while the expression of lncRNA CEBPA-AS1 in SNU-1 cells was the highest, and the differences were statistically significant (P<0.05). Compared with the si-NC group, the cell viability, the number of cell clone formation, migration and invasion, and the protein expression levels of MMP2 and MMP9 in the si-lncRNA CEBPA-AS1 group were significantly decreased, and the differences were statistically significant (P<0.05). Compared with the miR-NC group, the cell viability, the number of cell clone formation, migration and invasion, and the protein expression levels of MMP2 and MMP9 in the miR-455-3p group were significantly decreased, and the differences were statistically significant(P<0.05). lncRNA CEBPA-AS1 could target miR-455-3p and negatively regulate the expression of miR-455-3p. Compared with the si-lncRNA CEBPA-AS1+anti-miR-NC group, the cell viability, the number of cell clone formation, migration and invasion, and the protein expression levels of MMP2 and MMP9 in the si-lncRNA CEBPA-AS1+anti-miR-455-3p group were significantly increased, and the differences were statistically significant(P<0.05). Conclusion: Interference of the expression of lncRNA CEBPA-AS1 could inhibit the proliferation, clone formation, migration and invasion of gastric cancer cells through targeted regulating of miR-455-3p. |
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