| 刘文飞,秦洪真,董 翼,郭玉洋,胡文军,方 勇.环状RNA circMRPS35通过miR-130a-3p/ZNRF3轴调节胃癌细胞的增殖、凋亡、迁移和侵袭[J].,2022,(21):4020-4026 |
| 环状RNA circMRPS35通过miR-130a-3p/ZNRF3轴调节胃癌细胞的增殖、凋亡、迁移和侵袭 |
| Circular RNA circMRPS35 Regulates Proliferation, Apoptosis, Migration and Invasion of Gastric Cancer Cells through the miR-130a-3p/ZNRF3 Axis |
| 投稿时间：2022-06-08 修订日期：2022-06-30 |
| DOI：10.13241/j.cnki.pmb.2022.21.004 |
| 中文关键词: 胃癌 circMRPS35 microRNA-130a-3p 锌环指蛋白3 增殖 凋亡 迁移 侵袭 |
| 英文关键词: Gastric cancer circMRPS35 microRNA-130a-3p Zinc and ring finger 3 Proliferation Apoptosis Migration Invasion |
| 基金项目:北京市自然科学基金项目（71620715）；解放军第305医院院级课题（17YQ013） |
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| 中文摘要: |
| 摘要 目的：探讨环状RNA MRPS35（circMRPS35）对胃癌（GC）细胞增殖、凋亡、迁移和侵袭的调控机制。方法：体外培养人GC细胞系（HGC-27、MGC-803、MKN45和AGS）和正常胃上皮GES-1细胞，实时荧光定量PCR（RT-qPCR）检测circMRPS35、miR-130a-3p和锌环指蛋白3（ZNRF3）mRNA表达。另取MGC-803细胞，分为对照组、pc-NC组、pc-circMRPS35组、pc-circMRPS35+miR-NC组、pc-circMRPS35+miR-130a-3p组，采用Lipofectamine 3000进行质粒转染。RT-qPCR检测circMRPS35、miR-130a-3p和ZNRF3 mRNA表达，Western blot检测ZNRF3蛋白表达，CCK-8法、流式细胞术检测细胞增殖与凋亡，划痕实验和Transwell小室实验检测细胞迁移与侵袭能力，裸鼠移植瘤实验探究circMRPS35对GC细胞体内生长的影响。双荧光素酶报告基因检测miR-130a-3p与circMRPS35或ZNRF3的靶标关系。结果：GC细胞系中circMRPS35和ZNRF3 mRNA呈低表达，miR-130a-3p呈高表达（均P＜0.05）。过表达circMRPS35可降低miR-130a-3p，上调ZNRF3 mRNA和蛋白水平，抑制细胞增殖、迁移和侵袭，并促进细胞凋亡（均P＜0.05）；circMRPS35过表达对GC细胞恶性行为和裸鼠移植瘤生长的抑制作用可被miR-130a-3p mimic逆转（P＜0.05）。双荧光素酶实验结果显示，过表达miR-130a-3p可降低circMRPS35-WT和ZNRF3-WT的荧光素酶活性（P＜0.05）。结论：circMRPS35可能通过miR-130a-3p/ZNRF3轴抑制GC细胞的增殖、迁移和侵袭，并促进细胞凋亡。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the regulatory mechanism of circular RNA MRPS35 (circMRPS35) on the proliferation, apoptosis, migration and invasion of gastric cancer (GC) cells. Methods: Human GC cell lines (HGC-27, MGC-803, MKN45 and AGS) and normal gastric epithelial GES-1 cells were cultured in vitro, real-time quantitative PCR (RT-qPCR) was used to detect the mRNA expressions of circMRPS35, miR-130a-3p and zinc and ring finger 3(ZNRF3). MGC-803 cells were also taken and grouped into control group, pc-NC group, pc-circMRPS35 group, pc-circMRPS35 combined with miR-NC group, and pc-circMRPS35 combined with miR-130a-3p group. Plasmid transfection using Lipofectamine 3000. RT-qPCR was used to detect the mRNA expression of circMRPS35, miR-130a-3p and ZNRF3. Western blot was used to detect ZNRF3 protein expression, CCK-8 method and flow cytometry were used to detect cell proliferation and apoptosis, scratch assay and Transwell chamber assay were used to detect cell migration and invasion abilities, nude mouse xenograft experiments were performed to investigate the effect of circMRPS35 on the growth of GC cells in vivo. Dual-luciferase reporter genes were implemented to detect the target relationship of miR-130a-3p with circMRPS35 or ZNRF3. Results: In GC cell lines, the mRNA expression of circMRPS35 and ZNRF3 was low, and the expression of miR-130a-3p was high (all P<0.05). Overexpression of circMRPS35 was able to reduce the expression of miR-130a-3p, up-regulate the mRNA and protein levels of ZNRF3, inhibit cell proliferation, migration and invasion, and promote cell apoptosis (all P<0.05). The inhibitory effect of circMRPS35 overexpression on malignant behavior of GC cells and tumor growth in nude mice can be reversed by miR-130a-3p mimic (P<0.05). The results of dual luciferase experiment showed that overexpression of miR-130a-3p was able to reduce the luciferase activities of circMRPS35-WT and ZNRF3-WT (P<0.05). Conclusion: CircMRPS35 may inhibit the proliferation, migration and invasion of GC cells and promote apoptosis through the miR-130a-3p/ZNRF3 axis. |
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