【摘要】 目的观察PDCD5蛋白协同米非司酮(MIF)对前列腺癌细胞增殖及相关基因表达的影响。方法MTT法分别检测10、20、50和100μmol/L MIF作用于PC-3M细胞24~96 h的吸光度(A)值。扩增包含PDCD5序列的真核表达载体PCI-neo-PDCD5,用脂质体介导的方法转染前列腺癌PC-3M细胞。在转染PDCD5的细胞中分别加入10、20μmol/L MIF,继续培养24 h,MTT比色法检测细胞增殖,RT-PCR方法检测细胞增殖相关基因CyclinD1、Ki-67的表达。结果MTT实验表明,与对照组相比,10μmol/L MIF组的A值差异无统计学意义(P<0.05),20、50和100μmol/L MIF组的A值显著降低(P<0.05),MIF对前列腺癌PC-3M细胞的抑制作用呈剂量依赖性;PDCD5真核表达载体成功转染PC-3M细胞并得到了瞬时表达,转染PCI-neo-PDCD5并加入MIF后,与对照组及单独应用MIF组相比,细胞增殖受到明显抑制(P<0.05),增殖相关基因CyclinD1、Ki-67表达降低。结论PDCD5蛋白能够协同米非司酮抑制前列腺癌PC-3M细胞增殖,可能是通过调节某些增殖相关基因的表达来实现的,并有望成为前列腺癌的辅助治疗药物。更多还原

【Abstract】 ObjectiveTo investigate the effects of PDCD5 combined with mifepristone on proliferation of prostate cancer PC-3M cells.MethodsThe A values of the prostate cancer cells PC-3M that were treated with various concentrations(10,20,50,100 μmol/L) of MIF on time-course(24-96 h) were detected by MTT assay.The eukaryote expression vector PCI-neo-PDCD5 was amplified and transfected into PC-3M cells,and then the PC-3M cells were treated with 10 or 20 μmol/L MIF,the proliferation of the PC-3M cells was assayed by MTT,the expression of mRNA of CyclinD1,Ki-67 were detected by RT-PCR.ResultsThe A values of the cancer cells treated with 10 μmol/L of MIF were similar to those of controls(P<0.05),while those of the cells treated with 20,50 and 100 μmol/L of MIF were significantly different from those of controls(P<0.01).MIF markedly inhibited cell proliferation of prostate cancer PC-3M cells on a dose-and time-depending manner.The eukaryote expression vector PCI-neo-PDCD5 was successfully transfected and transient expressed in PC-3M cells.Compared with the control and MIF group,the viability of PC-3M cells treated with PDCD5 and MIF was decreased obviously,the expressions of CyclinD1,Ki-67 were down-regulated strikingly.ConclusionThe results suggest that PDCD5 may inhibit proliferation in PC-3M cells in combination with MIF.Its mechanism is maybe involved in the proliferation-related gene,including CyclinD1,Ki-67.更多还原