miR-572对胃癌细胞增殖和凋亡影响及其作用机制研究

目的 研究miR-572对胃癌细胞增殖、凋亡的影响和机制.方法 Real-time PCR方法 测定miR-572在胃癌细胞和正常胃黏膜细胞中的表达差异.胃癌细胞NCI-N87中转染miR-572 inhibitor,MTT测定细胞增殖,PI单染法测定细胞周期分布,Annexin V-FITC/PI法测定细胞凋亡变化,...

Effects and mechanism of miR-572 on proliferation and apoptosis of gastric cancer cells

Objective To study the effects and mechanism of micro RNA(miR)-572 on the proliferation and apoptosis of gastric cancer cells.Methods Differences in miR-572 expression between gastric cancer cells and normal gastric mucosal cells was detected by Real-time PCR.NCI-N87 cells transfected with the miR-572 inhibitor were examined using the following techniques:MTT assay of cell proliferation,propidium iodide(PI)single staining to determine cell cycle distribution,annexin V-fluorescein isothiocyanate/PI to measure apoptosis,and Western blot to detect the expression of cyclin-dependent kinase 4(CDK4),p21,B-cell lymphoma/leukemia-2(Bcl-2),and Bcl-2-associated X protein(Bax).An online target gene prediction software identified the gene encoding WW domain-containing oxidoreductase(WWOX)as a possible target of miR-572.This relationship was tested using a luciferase reporter system in which WWOX small interfering RNA(si RNA)and the miR-572 inhibitor were cotransfected into gastric cancer cells,followed by measurement of cell proliferation,cell cycle distribution and apoptosis as above.Results The expression level of miR-572 in gastric cancer cells was significantly higher than that in normal gastric mucosal cells.Gastric cancer cells transfected with the miR-572 inhibitor showed decreased proliferation,an increased proportion of G1 phase,an increased apoptosis rate,decreased expression of CDK4 and Bcl-2,and increased expression of p21 and Bax protein.,m i R-572 negatively regulated the expression of WWOX protein.WWOX si RNA reversed the inhibitory effect of the miR-572 inhibitor on proliferation,cell cycle arrest and apoptosis of gastric cancer cells.Conclusions Downregulation of miR-572-mediated negative regulation of WWOX inhibited the proliferation of gastric cancer cells,blocked the cell cycle and induced apoptosis.