微小RNA-143抑制食管癌细胞株ECA109的增殖、迁移和侵袭

Abstract

Objective To investigate the functions of microRNA-143 (miR-143) in esophageal cancer cell line ECA109. Methods ECA109 cells were transfected with negative control (NC), miR-143 mimics or miR-143 inhibitors. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium bromide (MTT) assay was performed to evaluate the growth of ECA109 cells after transfection. Annexin V-FITC/PI apoptosis test kit was used to detect early apoptosis rate in ECA109 cells. Transwell migration and invasion assays were conducted to compare the migration and invasion capacity of ECA109 among different groups. Real-time PCR and Western blotting were used to analyze the mRNA and protein alteration after transfection. Results Three and four days after transfection, compared with NC (absorbance value: 0.90±0.02 and 1.09±0.07), miR-143 mimics inhibited ECA109 cell proliferation (absorbance value: 0.66±0.05 and 0.80±0.04), while miR-143 inhibitors promoted cell proliferation (absorbance value: 1.13±0.09 and 1.51±0.08), with statistical significances (F=49.16, P=0.000; F=100.34, P=0.000). Early-stage apoptosis rates of ECA109 transfected with NC, miR-143 mimics and miR-143 inhibitors were 3.42%±0.72%, 11.63%±1.15% and 0.94%±0.10%, respectively, with statistical significance (F=151.61, P=0.000). Meanwhile, compared with NC (migration cell number: 336±13, invasion cell number: 147±16), miR-143 mimics inhibited cell migration (148±16) and invasion (75±10), while miR-143 inhibitors promoted cell migration (510±14) and invasion (238±16), with statistical significances (F=470.99, P=0.000; F=90.04, P=0.000). Compared with NC (1.00±0.00), miR-143 mimics down-regulated mRNA (relative expression level 0.22±0.08) and protein expression (relative expression level 0.46±0.08) of K-ras, whereas miR-143 inhibitors up-regulated mRNA (1.55±0.12) and protein expression (1.33±0.05) of K-ras (F=131.36, P=0.000; F=88.17, P=0.000). Conclusion miR-143 functions as a tumor suppressor in esophageal cancer cell line ECA109, probably by down-regulating K-ras expression. Key words: MicroRNAs; Esophageal neoplasms; Genes, tumor suppressor