Effects of circular RNA-Hsa-0101216 targeting microRNA-142-3p on proliferation, cloning, migration and invasion of pancreatic cancer cells
10.3760/cma.j.cn115667-20240424-00085
- VernacularTitle:环状RNA-Hsa-0101216靶向微小RNA-142-3p对胰腺癌细胞增殖、克隆、迁移及侵袭的影响
- Author:
Shaopeng LIU
1
;
Haichao LIU
1
;
Hongxian YAN
1
;
Minghui BAI
1
;
Jixiang ZHANG
1
Author Information
1. 洛阳市中心医院肝胆胰外科，洛阳 471000
- Publication Type:Journal Article
- Keywords:
Pancreatic cancer;
Cell proliferation;
Cell invasion;
Hsa-circ-0101216;
miR-142-3p
- From:
Chinese Journal of Pancreatology
2024;24(6):447-455
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of circular RNA-Hsa-0101216 (Hsa-circ-0101216) targeting microRNA-142-3p (miR-142-3p) on the proliferation,cloning,migration and invasion of pancreatic cancer cells, and explore its molecular mechanism.Methods:The differentially expressed miRNAs in pancreatic cancer were analyzed and screened with GEO database and the circRNA-miRNA network was constructed. The expression of Hsa-circ-0101216 miRNA and miR-142-3p in 5 strains of pancreatic cancer cells (BxPC-3, PANC1, MIA PaCa-2, Capan-2, CFPAC-1) and normal pancreatic duct epithelial cells (HPNE) was detected by quantitative real time PCR. The PANC1 and Capan-2 pancreatic cancer cells were divided into the Hsa-circ-0101216 small interference RNA transfection group (si-circRNA group), the senseless negative sequence siRNA transfection group (si-NC group), and the normal blank control (NC group); the miR-142-3p overexpression lentiviral vector transfection group (miR-142-3p mimic group), the empty vector transfection group (miR-CON group), the co transfection of Hsa-circ-0101216 siRNA and miR-142-3p mimic group (si-Hsa circ0101216+mimic miR-142-3p group), co transfection of Hsa-circ-0101216 siRNA and miR-142-3p mimic negative control sequence group (si-Hsa-circ-010126+mimic miR-142-3p-NC group). The changes of cell proliferation, cloning, migration and invasion were detected by CCK-8, EdU proliferative staining, plate cloning, cell scratch and Transwell assay. Detect the expression of ERK1 and ERK1 and ERK2 protein expression was measured by Western blotting, and the targeting relationship between Hsa-cic-0101216 and miR-142-3p was verified by dual luciferase reporter gene method.Results:The expression of miR-142-3p was significantly down-regulated by GEO analysis, and the Hsa-circ-0101216-miR-142-3p regulatory network was successfully constructed. The mRNA expression levels of Hsa circ0101216 in BxPC-3, PANC1, MIA PaCa-2, Capan-2, and CFPAC-1 cells were 5.64±0.34, 5.93±0.40, 5.66±0.14, 5.63±0.33, and 5.70±0.50, respectively, which were significantly higher than those in HPNE cells (1.27±0.06); the expression levels of miR-142-3p were 1.43±0.12, 1.20±0.09, 1.60±0.04, 1.16±0.25, and 1.42±0.11, respectively, which were significantly lower than those of HPNE cells (4.69±0.22), and all the differences were statistically significant (all P value <0.001). Among them, the expression level of Hsa-circ-0101216 mRNA in PANC1 cells was the highest, and the expression level in Capan 2 cells was the lowest. Compared with the NC group and si-NC group, the si-circRNA group showed a significant decrease in the absorbance value at 450 nm at 24, 48, and 72 hours ( A450 value), EdU positivity rate, cell clone number, migration rate, and transmembrane cell number of PANC1 and Capan-2 cells; the A450 value at the above time points, EdU positivity rate, cell clone number, migration rate, and transmembrane cell number of PANC1 and Capan-2 cells in the miR-142-3p mimic group were significantly lower than those in the NC group and miR-CON group. The A450 value, EdU positivity rate, cell clone number, migration rate, transmembrane cell number, ERK1 and ERK2 protein expression levels of PANC1 and Capan-2 cells in the si-Hsa-circ-0101216+mimic-miR-142-3p group were significantly reduced compared to the si-Hsa-circ-0101216+mimic miR-142-3p-NC group, and the differences were statistically significant (all P<0.001).The luciferase activity of PANC1 and Capan-2 cells in co transfected with wild-type (WT) Hsa-circ-0101216 and miR-142-3p mimic groups was significantly lower than that of the miR-CON+WT group (0.92±0.11 vs 2.33±0.21, 0.89±0.08 vs 2.30±0.17), and the differences were statistically significant (all P value <0.001), but there was no statistically significant difference on luciferase activity of PANC1 and Capan-2 cells between the co transfected mutant (MUT) Hsa-circ-0101216 and miR-142-3p mimic groups and the miR-CON+MUT group. Conclusions:Hsa-circ-0101216 is overexpressed in pancreatic cancer cells, while miR-142-3p is poorly expressed; Hsa-circ-0101216 can promote the proliferation, cloning, migration and invasion of pancreatic cancer cells by targeting miR-142-3p.
