Effect of microRNA-125a on invasion, migration and proliferation ability of human lung carcinoma H1975 cells by notch pathway

Abstract

Objective To investigate the effect and mechanism of microRNA (miRNA, miR)-125a in lung cancer cell invasion and migration. Methods MiR-125a-mimic was used to overexpress miR-125a. Polymerase chain reaction (PCR) was used to detect the effectiveness. Transwell invasion assay was used to detect invasion ability in miR-125a-mimic and normol control (NC); Scratch assay was used to detect migration ability in miR-125a-mimic and NC; cell counting kit-8 (CCK-8) was used to detect proliferation ability in miR-125a-mimic and NC; The expression of Notch-1, Notch intracellular domain (NICD)-1 and hairy and enhancer of splithomolog (Hes)-1 were detected by Western blotting; Luciferase reporter gene was used to detect interaction between miR-125a and Notch-1; Subcutaneous tumor was used to detect the size and volume of the tumor after inject the tumor cells. Immunohistochemistry was used to detect the expression of Notch-1. Results MiR-125a-mimic could overexpress miR-125a. miR-125a-mimic could inhibit the invasion ability of human lung carcinoma H1975 cells (0.22±0.03 vs. 0.89±0.13, P=0.008); miR-125a-mimic could inhibit the migration of human lung carcinoma H1975 cells; miR-125a-mimic could inhibit the proliferation of human lung carcinoma H1975 cells [24 h: (0.38±0.02)% vs. (0.19±0.01)%, P=0.025; 48 h: (0.72±0.05)% vs. (0.38±0.04)%, P=0.035; 72 h: (0.86±0.08)% vs. (0.43±0.04)%, P=0.019]; Overexpress the expression of miR-125a could reduce the expression of Notch-1, NICD-1 and Hes-1. Notch-1 is the direct target of miR-125a; compared with NC group, miR-125a-mimic group mice tumor volume and weight are significantly smaller [volume: (2.73±0.13)cm3 vs. (0.34±0.03) cm3,P=0.039; weight: (2.19±0.13) g vs. (0.36±0.03) g, P=0.028], immunohistochemistry showed that compared with NC group, the expression Notch-1 expression in miR-125a-mimic group decreased. Conclusion miR-125a can affect the invasion, migration and proliferation of lung carcinoma H1975 cells by regulating the pathway of Notch. Key words: MicroRNA-125a; Lung cancer; Notch-1; H1975; Invasion; Migration