吡柔比星联合塞来昔布对膀胱癌5637细胞株的作用

Effect of pirarubicin combined with celecoxib on bladder cancer cell line 5637

目的:体外观察吡柔比星(pirarubicin,THP)联合塞来昔布(celecoxib)对高危浅表性膀胱癌5637细胞株的增殖抑制作用及其可能机制。方法:MTT法检测不同浓度THP、塞来昔布以及2者间不同浓度组合对5637细胞的增殖抑制作用;FCM和Western印迹法检测THP、塞来昔布以及2药联合对5637细胞的诱导凋亡能力和对5637细胞质和细胞核中核因子κB(nuclear factor-κB,NF-κB)表达的影响。结果:MTT法检测结果显示,当低浓度(≤5μmol/L)THP联合塞来昔布时,能明显增加其对5637细胞的增殖抑制作用;FCM检测显示,100μmol/L塞来昔布、1μmol/L THP和2药联合应用,对5637细胞均无明显的诱导凋亡作用;Western印迹法检测显示,1μmol/L THP作用后,NF-κB大部分转入细胞核中,细胞质中仅有少量残留;当与100μmol/L塞来昔布联合应用后,NF-κB向细胞核内迁移的现象受到抑制。结论:膀胱癌5637细胞在THP作用后,细胞质内的NF-κB大部分转入细胞核中,联合塞来昔布后可以抑制这种迁移的现象,这可能是塞来昔布使THP药效增强的机制之一。

Objective：To observe the inhibitory effect of pirarubicin （THP） combined with celecoxib on proliferation of high-risk superficial bladder cancer cells （5637 cell line） in vitro and elucidate its possible mechanism. Methods：The inhibitory rate of 5637 cells after treatment with different concentrations of individual THP, celecoxib and THP plus celecoxib were observed by MTT assay. The cell apoptosis rate was determined by flow cytometry. The expression of nuclear factor-κB （NF-κB） in cytoplasms and nuclei was mea-sured by Western blotting.Results：MTT assay indicated that THP at low concentrations （≤5 μmol/L） combined with celecoxib can significantly enhance the inhibitory effect of THP on 5637 cell line. Flow cytometry showed that celecoxib 100 μmol/L and THP 1 μmol/L alone or their combination did not induce apparent apoptosis of 5637 cell line. Western blotting demonstrated that most NF-κB translocated from cytoplasm into nuclei after 1 μmol/L THP treatment. In contrast, the nuclear translocation of NF-κB in 5637 cell line was inhibited after combination treatment with THP and celecoxib. Conclusion：Most NF-κB translocated from cytoplasm into nucleus after 1 μmol/L THP treatment. Combined treatment with THP and celecoxib significantly inhibited the nuclear translocation of NF-κB, which could explain why celecoxib could enhance the efficacy of THP.