蛋白激酶抑制剂PKC412对膀胱癌T24细胞生长增殖的影响.docVIP

蛋白激酶抑制剂PKC412对膀胱癌T24细胞生长增殖的影响 　　作者：张建国1 ， 苏昀2　 作者单位：(1.江苏大学附属医院ICU， 江苏 镇江 212001; 2.东南大学附属中大医院泌尿外科， 江苏 南京 210009) 　　【摘要】 目的： 观察蛋白激酶C(PKC)抑制剂N苯甲酰星孢菌素(PKC412)对膀胱癌T24细胞株生长、增殖、侵袭功能和细胞周期的影响。方法： 用不同浓度PKC412(0.01,0.1,1.0,10 Μmol/L)作用于对数生长期T24细胞24,48,72 h。采用软琼脂克隆形成试验测定T24细胞克隆形成抑制率，MTT法检测细胞生长抑制率，Transwell小室法检测细胞的侵袭能力，流式细胞术分析细胞周期的变化。结果： 当PKC412浓度从0.01 Μmol/L增加到10 Μmol/L时，T24细胞克隆形成抑制率从-0.62%增加到80.46%，细胞生长抑制率从1.17%增加到73.62%，细胞侵袭抑制率从1.8%增加到73.72% (P0.05)，均呈时间和剂量依赖性。PKC412作用48 h后，随着浓度的增加，G2/M期细胞比例从12.26%增加到58.41%，呈G2/M期细胞阻滞。结论： PKC412对体外培养的T24细胞克隆形成、生长和侵袭具有抑制作用，其机制与G2/M期细胞阻滞有关。 　　【关键词】 膀胱癌;N苯甲酰星孢菌素;克隆;侵袭;细胞周期 　　[Abstract]　Objective： To investigate the growth ， proliferation， invasive function and the influence of the cell cycle of PKC412 on T24 bladder cancer in vitro. Methods： With different concentrations of PKC412 ( 0.01， 0.1， 1， 10 mol/L )acted on the T24 cell after 24， 48， 72 h; using soft agar clonogenic assay to determine the cell colony formation inhibition rate of T24， MTT method was used to detect the cell growth inhibition rate. The invasion ability of T24 was assayed by Transwell cell， the cell cycle changes were analysed by flow cytometric. Results： The inhibition rate of T24 cell clone forming from -0.62% increased to 80.46% ， when PKC412 concentration increased from 0.01 Μmol/L to 10 Μmol/L; cell growth inhibition rate increased from 1.17% to 73.62%， the cells invasive inhibition rate increased from 1.8% to 73.72%. All inhibitions were in a time-and-dose dependent manners. After PKC412 acting 48 hours， with the increasing of concentration， G2/M period cells from 12.26% increased to 58.41% ， showed the G2/M period cell block. Conclusion： PKC412 decreased cell growth，invasion and inhibited cloning formation of human bladder cancer cell T24 in vitro. 　　[Key words]　bladder cancer; N-benzoyl-staurosporine; clone; invasion; cell cycle 　　目前人们逐渐意识到化疗药物并不能使转移性膀胱癌的预后产生质的改变，因此学者们试图从基因和蛋白水平寻找诊断并治疗膀胱癌的可行性。N苯甲酰星孢菌素(N-benzoyl-staurosporine，PKC412)是十字孢碱(Staur