Effect and mechanism of miR-3188 on malignant biological behavior of gastric cancer cell
10.19405/j.cnki.issn1000-1492.2022.04.008
- Author:
Kunnan Wang
1
,
2
,
3
;
Jinming Zhang
3
,
4
;
Yun Zhang
3
,
4
;
Junze Zhang
3
,
4
;
Xinpu Yuan
3
,
4
;
Guijun Zou
3
,
4
;
Chaojun Zhang
1
,
2
,
3
Author Information
1. Naval Clinical College，Anhui Medical University，Hefei 230032
2. Dept of General Surgery， The Sixth Medical Centre，Chinese PLA General Hospital，Beijing 100048
3. Dept of General Surgery， The First Medical Centre，Chinese PLA General Hospital，Beijing 100853
4. Dept of General Surgery， The Sixth Medical Centre，Chinese PLA General Hospital，Beijing 100048
- Publication Type:Journal Article
- Keywords:
gastric cancer;
miR-3188;
NRAGE;
proliferation;
apoptosis;
invasion;
migration
- From:
Acta Universitatis Medicinalis Anhui
2022;57(4):545-551
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of miR-3188 on the proliferation, apoptosis, invasion and migration of gastric cancer cells and the underlying molecular mechanism.
Methods:The expression level of miR-3188 was examined in normal human gastric mucosal cells(GES-1) and human gastric cancer cell lines(HGC-27,MGC-803,BGC-823,MKN-45) by qRT-PCR. It was determined by bioinformatic analysis and dual luciferase reporter assay whether NRAGE was the target gene of miR-3188.miR-3188 mimic, miR-3188 inhibitor and negative control miR-NC were transfected into gastric cancer cell line HGC-27 respectively, and the expression of NRAGE on protein and mRNA levels was detected by Western blot and qRT-PCR. miR-3188 mimic, NRAGE overexpression plasmid(pc-NRAGE) and its negative control(miR-NC,pc-control) were separately or co-transfected into gastric cancer cell line HGC-27. CCK-8, flow cytometry and Transwell experiment were used to detect cell proliferation, apoptosis, invasion and migration in each group. The expressions of epithelial-mesenchymal transition(EMT), proliferation, apoptosis, invasion and migration related proteins [CyclinD 1,matrix metalloproteinase 9(MMP 9),Bcl-2,N-cadherin, Vimentin, E-cadherin] in each group were detected by Western blot.
Results:The expression of miR-3188 in human gastric cancer cell lines was lower than that of human normal gastric mucosal cells. Bioinformatics analysis and dual luciferase reporter assay confirmed that miR-3188 could target and bind to NRAGE 3′UTR.Western blot and qRT-PCR confirmed that miR-3188 negatively regulated the expression of NRAGE on the protein level in HGC-27 cells, but not on the mRNA level. Compared with the miR-NC group, Transfected miR-3188 mimic reduced the proliferation, invasion and migration ability of HGC-27 cells, and increased the apoptosis rate, and the co-transfection of pc-NRAGE could reverse the above effects. Compared with the miR-NC group, when miR-3188 was overexpressed in HGC-27 cell, the expression of cyclinD 1, MMP 9, Bcl-2, N-cadherin, and Vimentin was down regulated, while E-cadherin expression was up-regulated, and the above effects could be reversed by co-transfection with pc-NRAGE.
Conclusion:The miR-3188/NRAGE axis may play important roles in the progression of gastric cancer, and miR-3188 may be a potential therapeutic target for gastric cancer.
- Full text:2025021808592946211miR-3188对胃癌细胞...生物学行为的作用及机制研究_王坤男.pdf
