热化疗对膀胱癌BIU_87细胞侵袭运动能力的影响及相关机制

摘要

Objective To investigate the effect of thermo-chemotherapy by GEM on invasion ability of BIU-87 cells and its mechanism. Methods The logarithmic growth phase BIU-87 cells were divided into four groups,group A was the control group,group B was chemotherapy group, group C was thermo-chemotherapy group one [(40.0±0.2)℃],group D was the thermo-chemo-therapy group two [(43.0±0.2)℃].The invasion ability of the cells was detected by live cell mi-croscopy imaging system.The mRNA of MMP-2,MMP-9,RECK,TIMP-1,TIMP-2 were meas-ured by Real time-PCR experiment.The proteins of MMP-2,MMP-9,RECK,TIMP-1 and TIMP-2 were detected by ELISA. Results The live cell microscopy imaging system showed the invasion ability of the cells could be suppressed in group B,C,D compared with the control group,the inhib-itory effect of group D was the most obvious.Real time-PCR and ELISA experiments suggested that the mRNA and proteins of MMP-2,MMP-9 in group B,group C,group D were significantly less than that in group A (P<0.01),and group D was decreased most significantly.The RECK,TIMP-1 ,TIMP-2 in group B,group C,group D were significantly upregulated,compared with that in group A (P<0.01),and this upregulate in group D was the most significantly. Conclusions The thermo-chemotherapy of GEM suppress the invasion of BIU-87 cells by regulating the genes that cor-relate with invasive ability.%目的观察吉西他滨(gemcitabine,GEM)热化疗对膀胱癌BIU_87细胞侵袭运动能力的影响,探讨其可能的机制。方法将处于对数生长期的BIU_87细胞分为4组,A组为对照组,B组为单纯化疗组,C组为热化疗1组[(40.0±0.2)℃],D组为热化疗2组[(43.0±0.2)℃],GEM的工作浓度为1μg/ml。用活细胞显微成像系统实时监测细胞运动轨迹实验检测各组细胞的运动能力；用实时荧光定量PCR检测各组细胞的MMP_2、MMP_9、RECK、TIMP_1、TIMP_2 mRNA的表达情况；用ELISA法检测各组细胞的MMP_2、MMP_9、RECK、TIMP_1、TIMP_2蛋白的表达情况。结果活细胞显微成像系统实时监测细胞运动轨迹实验显示,相对于A组,B、C、D组对BIU_87细胞的运动能力均可产生抑制作用,其中以D组的抑制作用最为明显。实时荧光定量PCR及ELISA检测结果显示,与 A 组相比,B、C、D各组 MMP_2、MMP_9的 mRNA和蛋白均表达下调,而 RECK、TIMP_1、TIMP_2的 mRNA 及蛋白的表达均上调(P<0.01),其中以D组的变化最为明显。结论热化疗能够通过调节与侵袭相关基因的表达来抑制BIU_8 7细胞的迁移和侵袭,且具有温度依赖性。