人β防御素-1对膀胱癌T24细胞生长抑制作用的研究

Inhibitory effect of human beta-defensin-1 on bladder cancer T24 cells and its mechanism

目的探讨人β防御素-1(hBD-1)对膀胱癌T24细胞的生长抑制作用及机制。方法以不同浓度的hBD-1作用于体外培养的T24细胞,MTT法检测细胞生长抑制率,应用流式细胞仪观察测定细胞凋亡率,显微镜下观察细胞的形态学变化。结果 hBD-1对膀胱癌T24细胞株增殖抑制作用明显,与对照组(0μg/ml)比较,10～80μg/ml组在12、24、48h后均可抑制细胞增殖,差异均有统计学意义(t分别=25.24、37.20、56.49、55.81;25.02、44.92、48.46、55.08;26.79、23.63、30.54、35.54,P均<0.05)。4种浓度hBD-1组内各时点抑制率比较结果显示:12、24、48各浓度组内差异均有统计学意义(t分别=24.31、45.02、48.17、29.59、13.77、19.37;13.75、36.05、44.37、35.36、10.48、25.97;9.07、22.01、27.79、22.12、6.06、16.02,P均<0.05)。12h与24h间各浓度的比较,差异均有统计学意义(t分别=13.39、7.81、20.01、20.09,P均<0.05),但是24h与48h之间比较中10μg/ml与20μg/ml组比较,差异均有统计学意义(t分别=5.49、3.11,P均<0.05),其他各浓度组比较,差异均无统计学意义(t分别=2.09、1.65,P均>0.05)。组内各时点诱导T24细胞凋亡率比较结果显示:各浓度组12h与24h间的比较,差异均有统计学意义(t分别=7.07、7.18、11.43、9.75,P均<0.05),而在24h与48h间的比较,差异均无统计学意义(t分别=2.21、1.72、2.18、1.53,P均>0.05)。各浓度hBD-1处理组T24细胞形态均发生变化,表现为T24细胞肿大,体积增大,细胞边界模糊不清,胞质疏松,其内可见弥漫分布而大小不一空泡,出现细胞溶解现象,且随着hBD-1浓度增高凋亡细胞数量增多。结论 hBD-1对膀胱癌具有明显的体外抗肿瘤作用,细胞毒性作用可能是其重要作用机制。

Objective To investigate the inhibitory effect of human beta-defensin-1 on bladder cancer T24 cells and its mechanism.Methods Different concentrations of human beta-defensin-1 were added in culture medium.The inhibitory rate of the T24 cells was measured by MTT assay,cell apoptosis was observed by flowcytometry（FCM）.Cell morphology were observed by microscopy.Results Human beta-defensin-1 inhibited the growth of T24 cells significantly.Different concentrations groups compared to the control group（0 μg/ml）,with concentrations of human beta-defensin-1 increased,the suppression rate of the growth of T24 cells was increased too（t=25.24,37.20,56.49,55.81;25.02,44.92,48.46,55.08;26.79,23.63,30.54,35.54,P0.05）。At 12 hours,24 hours,48 hours,between different concentrations groups compared to each other,with concentrations of human beta-defensin-1 increased,the suppression rate of the growth of T24 cells was increased（t = 24.31,45.02,48.17,29.59,13.77,19.37;13.75,36.05,44.37,35.36,10.48,25.97;9.07,22.01,27.79,22.12,6.06,16.02,P0.05）。At 12 hours and 24 hours,the suppression rate of the growth of T24 cells was increased（t=13.39,7.81,20.01,20.09,P0.05）.But at 24 hours and 48 hours,the suppression rate was not so obvious,（10μg/ml and 20μg/ml groups t=5.49,3.11,P0.05;other groups t=2.09,1.65,P0.05）.At 12 hours and 24 hours,human beta-defensin-1 can also induced cell cytolysis（t= 7.07,7.18,11.43,9.75,P0.05）;But at 24 hours and 48 hours,this phenomenon rate was not so obvious（t=2.21,1.72,2.18,1.53,P0.05）。Different concentrations of human beta-defensin-1 can result T24 change in morphology,the cell volume inceased,the boundary of cells were not clear,cytokinesis became loose,in which disseminate vacuole,that was called cytolysis,and it was also in a dose-dependent manner.Conclusions Human beta-defensin-1 has inhibitory effect on bladder cancer T24 cells in vitro,which might be associated with the cell cytotoxicity.