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.
ABSTRACT
Objective To observe the regulation of microRNA (miRNA,miR)-758-3p on the expression of murine double microsomal gene 2 (MDM2) and its effect on invasion and proliferation of gastric cancer cell line MGC803.Methods The bioinformatics software was used to predict MDM2 as target gene of miR-758-3p.The wild type MDM2 gene 3'untranslated region luciferase reporter gene vector and miR-758-3p target sequence mutated vector and the corresponding miRNA were transfected into gastric cancer cells MGC803 by lipofectamine.Dual luciferase reporter system detects luciferase activity.The miR-758-3p mimics were transfected into gastric cancer cell MGC803 by lipofectamine.Real-time PCR was used to detect the transfection efficiency.Real-time PCR and Western blot were used to detect miR-MDM2 expression level in cells after transfection.Transwell assay and CCK-8 assay were used to detect cell invasion and proliferation.SPSS 20.0 was used to conduct the statistical analysis.Results Dual luciferase reporter assay confirmed that miR-758-3p could target MDM2 gene(P ＜ 0.05).The expression level of miR-758-3p in MGC803 cells transfected with miR-758-3p mimics was significantly higher than that in miR-NC cells [(6.68 ±0.53) vs (0.84 t0.12),P ＜0.01].Compared with miR-NC group,MDM2 expression was down-regulated in MGC803 cells transfected with miR-758-3p mimics (P ＜ 0.05).The number of invasive cells in miR-NC group and miR-758-3p group were (136.00 ± 16.62) and (79.49 ± 6.42).After knockdown MDM2,the invasiveness of cells was significantly decreased (P ＜ 0.05).The results of CCK-8 showed that the proliferation of MGC803 cells transfected with miR-758-3p group was significantly lower than that of miR-NC group (P ＜ 0.01).Conclusion miR-758-3p can reduce the invasion and proliferation of MGC803 cells by targeting MDM2.