6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶4对癌细胞的增殖及迁移的影响及机制研究

PFKFB4 Promotes the Proliferation and Migration of Ovarian Cancer Cells Through Regulating Glycolysis

目的:探讨6-磷酸果糖-2-激酶/果糖-2,6-二磷酸酶4(PFKFB4)对卵巢癌细胞增殖和迁移的影响及其机制。方法:利用慢病毒构建PFKFB4沉默和过表达的卵巢癌A2780稳定细胞系,并用qPCR和Western blotting检测PFKFB4沉默和过表达效果,通过CCK-8实验、克隆形成实验、划痕愈合实验、Transwell实验、流式细胞术和试剂盒分别检测A2780细胞的增殖和迁移能力以及糖酵解水平。结果:成功构建了PFKFB4沉默和过表达的A2780稳定细胞系。与对照组相比,沉默PFKFB4后,A2780细胞的PFKFB4 mRNA和蛋白水平、增殖和迁移能力以及葡萄糖消耗和乳酸分泌水平显著降低,ROS水平显著升高,差异有统计学意义(P<0.05);过表达PFKFB4后,A2780细胞的PFKFB4 mRNA和蛋白水平、增殖和迁移能力以及葡萄糖消耗和乳酸分泌水平显著升高,ROS水平显著降低,差异有统计学意义(P<0.05)。结论:PFKFB4可通过增强糖酵解活性,发挥促进卵巢癌细胞的增殖和迁移的作用。

Objective To investigate the effects and mechanism of 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase-4(PFKFB4)on the proliferation and migration of ovarian cancer cells.Methods PFKFB4 knockdown and overexpression A2780 stable cell lines were established using lentivirus.The knockdown and overexpression efficacy of PFKFB4 was detected by qPCR and Western blotting.The proliferation,migration and glycolysis level of A2780 cells were detected by CCK-8 assay,clone formation assay,scratch healing assay,Transwell assay,flow cytometry and kits respectively.Results A2780 stable cell lines with PFKFB4 knockdown and overexpression were successfully established.Compared with the control group,PFKFB4 mRNA and protein levels,proliferation and migration ability,glucose consumption and lactic acid secretion levels of A2780 cells were significantly decreased after PFKFB4 knockdown(P<0.05),and ROS level was significantly increased(P<0.05).After overexpression of PFKFB4,the levels of PFKFB4 mRNA and protein,proliferation and migration ability,glucose consumption and lactic acid secretion of A2780 cells were significantly increased(P<0.05),and the ROS level was significantly decreased(P<0.05).Conclusion PFKFB4 promotes the proliferation and migration of ovarian cancer cells through glycolysis.