紫铆因抑制膀胱癌细胞增殖的体内外研究

Butein Inhibits Cell Proliferation of Bladder Cancer in vitro and in vivo

目的观察紫铆因对膀胱移行细胞癌细胞BLS增殖的影响以及对膀胱癌裸鼠移植瘤生长的影响。方法不同浓度紫铆因处理细胞,四甲基偶氮唑蓝比色实验(MTT)及平板克隆形成实验观察细胞增殖能力,流式细胞仪分析细胞周期,蛋白印迹检测核转录因子κB(NF-κB)p65核内表达及细胞外信号调节激酶1/2(ERK1/2)的磷酸化程度,并检测NF-κB下游靶基因细胞周期素D1(Cyclin D1)及环氧合酶-2(COX2)的表达。建立膀胱癌裸鼠皮下移植瘤,采用腹腔注射的给药途径,测量移植瘤的体积和重量。结果紫铆因抑制了膀胱癌细胞增殖并诱导G2/M期细胞周期阻滞。紫铆因处理后NF-κB p65的核内表达及ERK1/2的磷酸化程度下降(P<0.05),Cyclin D1及COX2基因表达下调(P<0.05),体内实验发现紫铆因治疗组较对照组皮下移植瘤的生长明显受抑制(P<0.05)。结论紫铆因具有抗膀胱癌细胞增殖作用,可能与其抑制ERK及NF-κB信号激活有关。

Objective To investigate the effects of Butein on proliferation of BLS cells in human bladder transitional cell carcinoma(BTCC) in vitro and on transplantation tumor growth of bladder carcinoma in nude mice. Methods BLS cells were treated with Butein at various concentrations. Cell proliferation was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay and clone formation assay, and cell cycle was detected by flow cytometry. Western blot was applied to measure phosphorylation of extra cellular signal regulated kinase(ERK1/2), nuclear transcription factor kappa B(NF-κB) p65 expression, Cyclin D1 and COX2 expressions in downstream target genes of NF-κB. Transplanted tumors of bladder cancer in nude mice were constructed. Administration route was intraperitoneal injection. Volume and weight of transplanted tumors were measured. Results Butein inhibited cell growth and induced G2/M cell cycle arrest in bladder cancer cells. Phosphorylation of ERK1/2 and NF-κB p65 intranuclear expression were reduced(P<0.05); Protein expressions of Cyclin D1 and COX2 were decreased(P<0.05); Growth of transplant subcutaneous tumor in treatment group was obviously inhibited(P<0.05). Conclusion Butein has anti-proliferation effect on human bladder cancer cells in vitro and in vivo possibly through suppressing ERK and NF-κB activation.