摘要
目的:探索异丹叶大黄素(ISO)抑制膀胱癌细胞侵袭、转移和增殖活性的机制。方法:以ISO作用于人源性膀胱癌UMUC3细胞后,倒置相差显微镜下观察并以ATP生物荧光法检测癌细胞增殖活性;以RT-PCR和Western blotting法检测细胞周期蛋白D1表达;以流式细胞术检测细胞周期的变化;细胞划痕法检测细胞迁移活性。结果:20μmol/L浓度以上ISO可显著抑制UMUC3细胞的增殖,其IC50为(22.5±2.8)μmol/L。同时UMUC3细胞的细胞周期蛋白D1 mRNA和蛋白水平均显著降低。以低浓度5μmol/L ISO预处理UMUC3细胞后,与对照组(47.33%)进行比较,可分别在12 h(58.82%)和24 h(63.94%)显著诱导细胞G0/G1期阻滞(P<0.01)。细胞划痕法检测证实5μmol/L ISO可显著抑制膀胱癌细胞的迁移活性。结论:ISO可抑制膀胱癌细胞增殖,下调细胞周期蛋白D1表达,诱导G0/G1细胞周期阻滞,抑制细胞的迁移能力。
AUM： To explore the inhibitory mechanism of isorhapontigenin （ISO） on the proliferation, migra- tion and invasion of UMUC3 bladder cancer cells. METHODS ： Human UMUC3 bladder cancer cells were pretreated with ISO, and the proliferation of the cells was observed under phase-contrast microscope and by ATPase assay. The expression of cyclin D1 was determined by RT-PCR and Western blotting. The cell cycle alteration was detected by flow cytometry, and the cell migration was examined by wound-healing assay. RESULTS： Over 20 p^mol/L of ISO significantly inhibited the proliferation of UMUC3 cells with the ICso of （22.5 ＋ 2.8） /xmol/L. The mRNA and protein levels of cyclin D1 in UMUC3 cells were markedly decreased after treatment with ISO. Exposure of UMUC3 cells to low dose （5 p^mol/L） of ISO led to significant induction of G0/Gl growth arrest at both 12 h （58.82%） and 24 h （63.94%）, compared with the nega- tive control cells （47.33%） without inducing obvious apoptosis. ISO at dose of 5 /xmol/L also markedly inhibited the cell migration. CONCLUSION： ISO significantly exhibits inhibitory effects on the proliferation and migration of human bladder cancer cells by down-regulation of cyclin D1 expression accompanying with G0/Gl cell cycle arrest.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2013年第3期442-448,共7页
Chinese Journal of Pathophysiology
基金
浙江省中医药青年基金资助项目(No.2005-A-12)
