【摘要】 目的研究尿激酶型纤溶酶原激活物(uPA)和ATP动力型质子泵(V-ATPase)在肝癌细胞株的表达,及抑制V-ATPase与癌细胞抑制和凋亡的关系。方法用RT-PCR和Western blot检测稳定转染真核表达质粒pCMV-uPA-HA的HCC97L细胞及转染空载体pCMV-HA的HCC97L细胞中uPA和V-ATPase mRNA和蛋白的表达;在转染uPA的HCC97L细胞培养基中加入不同浓度的Bafilomycin A1,用流式细胞仪和CCK-8分别检测癌细胞凋亡及受抑制情况。结果将uPA转染入低转移潜能肝癌细胞株HCC97L后,uPA mRNA和蛋白表达强于转染空载体的HCC97L,V-ATPase mRNA和蛋白表达随着转染uPA而增强。在细胞培养基中加入Bafilomycin A1后细胞生长增殖受到抑制、凋亡增加,且抑制率和凋亡率均随Bafilomycin A1浓度的加大而增加。结论 V-ATPase mRNA和蛋白表达随癌细胞转移能力的提高而增强;Bafilomycin A1抑制V-ATPase,干扰癌细胞内H+泵出,从而抑制癌细胞生长增殖、促进癌细胞凋亡。更多还原

【Abstract】 Objective To study the expression of urokinase type plasminogen activator(uPA) and vacuolar-type H+-ATPases(V-ATPases) in the hepatocellular carcinoma cell lines(HCC97L) and their relationship with inhibition and apoptosis of cells when V-ATPase was inhibited.Methods mRNA and protein expression of uPA and V-ATPase in uPA-transfected HCC97L were detected by RT-PCR and Western blot methods.Apoptosis rate and inhibition rate of uPA-transfected HCC97L were detected by using flow cytometer and cell counting Kit-8(CCK-8) respectively when different concentration Bafilomycin A1 was put into the culture medium.Results Both mRNA and protein expression of uPA in uPA-transfected HCC97L were higher than those of the vector-transfected ones;the expression of V-ATPase coincided with uPA’s.The growth and proliferation were inhibited and apoptosis was increased when cancer cells treated with Bafilomycin A1.Conclusions mRNA and protein expression of V-ATPase increased along with the increase of cell metastasis abilitiy.The proliferation and growth were suppressed and the apoptosis was increased in the hepatocellular carcinoma cells when V-ATPase was inhibited by Bafilomycin A1.更多还原