蟾毒灵对黑色素瘤A375细胞的作用及其机制研究  

Effect of bufalin on A375 melanoma cells and its mechanism

作　　者：任煜[1] 刘婷婷[1] 祁冰洁 陈玉 Ren Yu;Liu Tingting;Qi Bingjie;Chen Yu(Anqing Medical College,Anqing 246052,Anhui,China)

机构地区：[1]安庆医药高等专科学校,安徽安庆246052

出　　处：《右江民族医学院学报》2021年第6期719-724,共6页Journal of Youjiang Medical University for Nationalities

基　　金：安徽省高校优秀拔尖人才培育资助项目(gxyqZD2020060);安庆医药高等专科学校自然科学研究项目(2020ZR007)。

摘　　要：目的 探讨蟾毒灵(Bufalin)对黑色素瘤A375细胞的作用及其机制研究。方法 体外培养黑色素瘤A375细胞,采用CCK-8法检测不同浓度的蟾毒灵对A375细胞增殖的影响;采用Annexin V-FITC/PI双染,流式细胞仪检测和细胞周期分布情况;JC-1法检测蟾毒灵对A375细胞对线粒体膜电位的影响;采用分光光度法检测Caspase-9、Caspase-3蛋白的活性。结果 不同浓度蟾毒灵(6.25nmol/L、12.5nmol/L、25nmol/L、50nmol/L、100nmol/L、200nmol/L、400nmol/L)作用于A375细胞24h、48h和72h后,IC50值分别为107nmol/L、85.18nmol/L、41.62nmol/L,IC50值逐渐下降,实验组与空白对照组比较,随着蟾毒灵浓度的增加和作用时间延长,A375细胞活力均有显著下降,说明蟾毒灵可明显抑制A375细胞增殖,且呈浓度依赖性和时间依赖性;蟾毒灵可显著增加A375细胞凋亡率并阻滞细胞周期于S期;明显降低A375细胞线粒体膜电位;蟾毒灵可明显增加A375细胞内Caspase-9、Caspase-3蛋白的活性。结论 蟾毒灵可抑制黑色素瘤A375细胞增殖,诱导A375细胞凋亡,其作用机制可能与激活Caspase家族有关。Objective To investigate the effect of bufalin on A375 melanoma cells and its mechanism.Methods A375 melanoma cells were cultured in vitro,and CCK-8 assay was employed to detect the effect of bufalin at different concentrations on A375 cell proliferation.Annexin V-FITC/PI double staining was performed,and cell-cycle distribution was detected by flow cytometry.JC-1 was used to detect the effect of bufalin on the mitochondrial membrane potential of A375 cells.The protein activity of Caspase-9 and Caspase-3 was detected by spectrophotometry.Results A375 cells were treated with bufalin at different concentrations(6.25 nmol/L,12.5 nmol/L,25 nmol/L,50 nmol/L,100 nmol/L,200 nmol/L,400 nmol/L)for 24 h,48 h and 72 h.Their IC50 values were 107 nmol/L,85.18 nmol/L and 41.62 nmol/L,respectively.The IC50 values gradually decreased.Compared with the blank control group,the experiment group had a significant decrease in the viability of A375 cells,with the increase of bufalin concentration and the treatment time.These results indicated that bufalin could significantly inhibit the proliferation of A375 cells in a concentration-dependent and time-dependent manner.Bufalin could significantly increase the apoptosis rate of A375 cells and block the cell cycle in S phase.It could also significantly reduce the mitochondrial membrane potential of A375 cells,as well as increased the activity of proteins Caspase-9 and Caspase-3 in A375 cells.Conclusion Bufalin can inhibit the proliferation of A375 melanoma cells and induce their apoptosis.Its mechanism may be related to the activation of Caspase family.

关 键 词：黑色素瘤 A375细胞 蟾毒灵 细胞凋亡 

分 类 号：R965[医药卫生—药理学] R739.5[医药卫生—药学]