蟾毒灵通过下调PKM2抑制黑色素瘤细胞增殖、迁移与侵袭的实验研究

Bufalin Inhibits the Proliferation, Migaration and Invasion of Melanoma Cells by Down-regulating PKM2

目的:考察蟾毒灵(Bufalin)对黑色素瘤细胞增殖、迁移与侵袭,以及对有氧糖酵解关键酶M2型丙酮酸激酶(pyruvate kinase M2,PKM2)的调控作用。方法:分别使用MTT、划痕及Transwell实验,考察蟾毒灵对A375黑色素瘤细胞增殖、迁移与侵袭的影响;利用试剂盒检测蟾毒灵对A375细胞有氧糖酵解主要指标乳酸、葡萄糖及丙酮酸激酶的影响;蛋白免疫印迹实验探讨蟾毒灵对PKM2蛋白表达的影响,以及对下游肿瘤迁移与侵袭相关蛋白表达的影响。结果:蟾毒灵能显著抑制A375细胞的增殖、迁移与侵袭;还能抑制A375细胞的有氧糖酵解及PKM2的蛋白表达,进而下调肿瘤转移相关蛋白MMP-2、MMP-9、N-cadherin的表达。结论:蟾毒灵对A375黑色素瘤细胞的增殖、迁移与侵袭具有明显的抑制作用,其抑制机制可能与抑制肿瘤有氧糖酵解途径中关键酶PKM2有关。

Objective: To investigate the effects of bufalin on the proliferation, migration and invasion of melanoma cells, and its effect on the key enzyme of aerobic glycolysis pyruvate kinase M2 (PKM2). Methods: The effects of bufalin on the proliferation, migration and invasion of A375 melanoma cells were observed by MTT, scratches and Transwell methods, respectively. The effects of bufalin on the main indicators of aerobic glycolysis (including lactic acid, glucose and pyruvate kinase) were tested by related activity kits. The effects of bufalin on the expression of PKM2 and other proteins related to migration and invasion were tested by Western blotting. Results: Bufalin significantly inhibited the proliferation, metastasis and invasion of A375 melanoma cells;Bufalin could inhibit the aerobic glycolysis of A375 melanoma cells, and could suppress the enzyme activity of PKM2 significantly. Moreover, bufalin could inhibit the protein expressions of PKM2, MMP2, MMP9 and N-cadherin. Conclusions: Bufalin exerts the inhibitory effect on the proliferation and invasion of A375 melanoma cells, and the mechanism is, at least partly, due to the inhibition of PKM2 in tumor aerobic glycolysis.