目的: 探索葛根素抑制膀胱癌(bladder cancer, BLCA) T24及5637细胞系的作用机制。方法: 利用Cell Counting Kit-8(CCK-8)检测法证实葛根素抑制BLCA T24及5637细胞的能力。通过串联质谱标签(Tandem Mass Tags, TMT)技术获得差异表达蛋白列表,并进行功能富集分析。通过生物信息分析筛选关键蛋白并对其进行表达分析、生存分析和上游转录因子预测。分子对接及Western blotting实验用于上游转录因子的验证。结果: CCK-8检测结果显示葛根素对T24和5637细胞系的IC₅₀分别为218 μmol·L^-1和198 μmol·L^-1。功能富集分析显示差异表达蛋白主要富集在细胞周期和DNA复制通路。筛选出的关键蛋白为CDK1、CCNB1和PLK1。CHEA3网站预测关键蛋白上游转录因子为着丝粒蛋A(centromere protein A,CENPA。分子对接显示葛根素与CENPA的结合能为-6.3 kcal·mol^-1(1 cal=4.184 0 J),Western blotting显示葛根素可显著降低CENPA的表达。结论: 葛根素可能通过抑制CENPA的表达来影响蛋白CCNB1和PLK1,从而调控BLCA细胞的细胞周期或DNA复制以抑制其增殖。

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