Effect of miR-7856-5p on migration and proliferation of colorectal cancer cell line SW480 by targeting EPHA3
10.3760/cma.j.cn115396-20190908-00160
- VernacularTitle:miR-7856-5p通过靶向作用 EPHA3对结直肠癌细胞SW480迁移和增殖的影响
- Author:
Mingming DONG
1
;
Geng HUANG
;
Shujiang PENG
;
Xinming LI
;
Youneng YUAN
Author Information
1. 鄂东医疗集团黄石市中心医院(湖北理工学院附属医院)胃肠外科 435000
- From:
International Journal of Surgery
2020;47(4):226-231
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the regulation of microRNA (miRNA, miR)-7856-5p on the expression of EPH receptor A3 (EPHA3) gene and its effect on the migration and proliferation of colorectal cancer cell line SW480.Methods:Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression of miR-7856-5p in colorectal cancer tissues and cell lines. The miR-7856-5p mimic and miR-NC were transfected into colorectal cancer cell line SW480 by lipofection, respectively, and defined as miR-7856-5p group and miR-NC group, respectively. Real-time PCR was used to detect transfection effects. Transwell assay and CCK-8 assay were used to detect cell migration and proliferation after transfection. The bioinformatics software predicts and the dual luciferase reporter gene system validates the target gene of miR-7856-5p. Real-time PCR and Western blot were used to detect the expression of EPHA3 in the transfected cells. The measurement data in accordamce with normal distribution were expressed as mean±standard deviation, t test was used for inter grap comparison, and single factor analysis of variance was used for multi group comparison. Results:The expression of miR-7856-5p in colorectal cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). The expression of miR-7856-5p in colorectal cancer cell lines was significantly lower than that in normal intestinal mucosal epithelial cells ( P<0.05), and lowest in SW480 cells ( P<0.01). The expression of miR-7856-5p in miR-7856-5p group was significantly higher than that in miR-NC group, the difference was statistically significant [(9.49 ± 1.09) vs (1.06 ±0.18), P<0.01]. The number of migrated cells in miR-NC group and miR-7856-5p group were (125.70±14.05) and (42.01±8.98), respectively, and the migration ability of miR-7856-5p group was significantly decreased ( P<0.01). Compared with the miR-NC group, the cell proliferation ability of the miR-7856-5p group was significantly decreased ( P<0.05). Bioinformatics software showed that the target gene for miR-7856-5p was EPHA3. The dual luciferase reporter gene system confirmed that miR-7856-5p can target the EPHA3 gene ( P<0.05). Compared with the miR-NC group, the expression of EPHA3 in the SW480 cells of the miR-7856-5p group was significantly decreased ( P<0.05). Conclusion:miR-7856-5p can inhibit the migration and proliferation of colorectal cancer cell SW480 by regulating the expression of EPHA3.
