Silencing PKM2 by siRNA Inhibits Proliferation and Glycolysis of Human Gastric Adenocarcinoma Cell Line SGC-7901 / 胃肠病学

ABSTRACT

Background:Pyruvate kinase M2( PKM2 ),which is highly expressed in cancer cells,plays an important role in cancer cell growth and aerobic glycolysis and is a promising target for cancer therapy. Aims:To investigate the effect of silencing PKM2 by siRNA on proliferation,apoptosis,migration,invasion and glycolysis of gastric cancer cell line SGC-7901 cells. Methods:SGC-7901 cells were divided into three groupsnon-transfectd SGC-7901 cells,SGC-7901 cells transfected with empty plasmid and SCG-7901 cellls transfected with PKM2 siRNA. Expression of PKM2 was detected by fluorescent quantitative PCR and Western blotting at mRNA and protein levels. Intracellular expression and distribution of PKM2 was determined by immunofluorescence. CCK-8 assay,flow cytometry,migration and invasion experiment were used to assess cell proliferation,apoptosis,migration and invasion,respectively. Western blotting was used to determine the expression of glucose transporter-1( Glut-1)and lactate dehydrogenase A( LDHA). Spectrophotometry was used to detect levels of extracellular glucose and lactic acid and intracellular lactate dehydrogenase( LDH). Results:Compared with non-transfected group and negative control( empty plasmid )group,PKM2 mRNA and protein expressions were significantly decreased in PKM2 siRNA plasmid transfected group(P<0. 05). Intracellular expression of PKM2 was markedly decreased in PKM2 siRNA cells. After transfected with PKM2 siRNA plasmid,cell proliferation,migration and invasion were significantly inhibited,and cell apoptosis was increased significantly( P<0. 05 ). Expressions of Glut-1 and LDHA were down-regulated in PKM2 silenced cells( P <0. 05 ). Glucose uptake,lactic acid production and LDH activities were significantly decreased in PKM2 silenced cells(P<0. 05). Conclusions:RNA interference targeting PKM2 gene inhibits PKM2 expression in human gastric adenocarcinoma cell line SGC-7901 cells,thereby inhibits cell proliferation and glycolysis.