Targeted regulation of microRNA?150?3p on IGF1 expression and its effect on invasion and migration of gastric cancer cells
YANG Chaowen
DING Chengming
GUI Pucheng
YU Zijian
Abstract：Objective To investigate the targeted regulation of microRNA?150?3p (miR?150?3p) on insulin like growth factor 1 (IGF1) expression and its effect on invasion and migration of gastric cancer cells. Methods The miR?150?3p levels in normal gas?tric mucosal cell GES1 and gastric cancer cell lines AGS and MGC803 were detected by quantitative fluorescence PCR ( QPCR). MGC803 cells were transfected with miR?150?3p mimics (Overexpression group) or inhibitors (Inhibition group) by liposome method, and cells transfected with negative control sequence were used as Control group. The levels of miR?150?3p and IGF1 were detected by QPCR at 24 h after transfection to evaluate the transfection efficiency. Scratch healing rate and number of proliferating cells were detec?ted by scratch test and Transwell test, respectively. The levels of IGF1, phosphorylated protein kinase B (p?Akt) and phosphorylated phosphoinositide 3?kinase (p?PI3K) were detected by Western blotting. The targeting relationship and binding site between miR?150?3p and IGF1 were detected by double luciferase reporter gene assay. Results The levels of miR?150?3p in AGS and MGC803 cells were 3. 711±0. 726 and 4. 861±0. 854, higher than 1. 025±0. 265 in GES1 cells ( P＜0. 05). The miR?150?3p level in Control group was 1. 045±0. 574, while that increased to 4. 168±0. 897 in Overexpression group and decreased to 0. 247±0. 097 in Inhibition group (P＜0. 05). The scratch healing rate and the number of penetrating cells in Control group were (54. 5±3. 2)% and 195. 2±12. 8, while those increased to (75. 2±4. 1)% and 274. 6±15. 9 in the Overexpression group and decreased to (21. 5±2. 1)% and 124. 7±10. 6 in Inhibition group (P＜0. 05). Compared with the Control group, the levels of p?PI3K and p?Akt in the overexpression group increased, while those in the Inhibition group decreased (P＜0. 05). MiR?150?3p mimics could inhibit the relative luciferase activity in the 3’ un? translated region of wild?type IGF1 (P＜0. 05), but had no effect on the mutant (P＞0. 05). The mRNA and protein levels of IGF1 in Control group were 1. 025±0. 086 and 0. 458±0. 073, while those reduced to 0. 341±0. 059 and 0. 147±0. 081 in Overexpression group (P＜0. 05). Conclusion MiR?150?3p is highly expressed in gastric cancer cells. Downregulation of miR?150?3p can inhibit migration and invasion and activation of PI3K／Akt pathway. It may play a role in promoting cancer by targeting IGF1. This miRNA may be a po?tential target for diagnosis and treatment of gastric cancer.
Keywords：Gastric cancerMicroRNA?150?3pInvasive and migrationInsulin like growth factor 1(IGF1)
Publication Date：2019-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 769-773 )