TFAR19蛋白协同米非司酮对前列腺癌PC-3M细胞凋亡的影响

目的初步探讨TFAR19协同米非司酮(MIF)对前列腺癌PC-3M细胞凋亡的影响。方法构建TFAR19真核表达载体,用脂质体介导的方法转染PC-3M细胞。MTT法检测5、10、20、50和100μmol·L-1MIF作用于前列腺癌PC-3M细胞24~96h的吸光度(A)值。在转染TFAR19的细胞中加入20mol·L-1MIF培养24、48h,MTT比色法检测细胞增殖,原位末端标记(TUNEL)法检测细胞凋亡率,透射电镜进一步观察细胞超微结构的改变。结果构建了PCI-neo-TFAR19真核表达载体并在转染的PC-3M细胞中得到了瞬时表达。MTT实验表明,与对照组相比,5、10μmol·L-1MIF组的A值差异无统计学意义(P>0.05),20、50和100μmol·L-1MIF组的A值差异有统计学意义(P<0.01),MIF对前列腺癌PC-3M细胞的抑制作用呈时间、剂量依赖性;转染PCI-neo-TFAR19并加入20 mol·L-1MIF后,与对照组及单独应用MIF组相比,细胞生长明显受到抑制(P<0.01),细胞凋亡率明显增加(P<0.01),透射电镜观察到典型的细胞凋亡特征(细胞体积缩小,核皱缩、碎裂,染色质呈块状边集等)。结论TFAR19蛋白能够协同米非司酮促进前列腺癌PC-3M细胞凋亡,有望成为前列腺癌的辅助治疗药物。

Cooperation of protein TFAR19 and mifepristone on apoptosis of human prostate cancer cells PC-3M

Aim To study the cooperation effect of TFAR19 protein and mifepristone on apoptosis of prostate cancer cells PC-3M.Methods The eukaryote expression vector of TFAR19 was constructed and transfected into PC-3M cells by liposome.The A values of the prostate cancer cells PC-3M in each group with various concentrations(5,10,20,50,100 μmol·L-1) of MIF at various time intervals(24～96 h) were detected by MTT assay.PCI-neo-TFAR19 was transfected into PC-3M cells treated with 20 μmol·L-1 MIF,the proliferation of PC-3M cells was assayed by MTT,TUNEL(situ end-labeling) was used to detect the rate of apoptosis.The change of morphology of apoptotic cells was observed by electron microscope.Results The eukaryote expression vector of TFAR19 gene was constructed and transiently expressed in PC-3M cells.The A values of the cancer cells treated with 5,10 μmol·L-1 of MIF were similar to those of controls,while those of the cells treated with 20,50 and 100 μmol·L-1 of MIF were significantly different from those of controls(P<0.01).MIF markedly inhibited cell proliferation of prostate cancer cells PC-3M on a dose-and time-dependent manner.Compared with the control and MIF group,the result of MTT showed that the viability of PC-3M cells treated with TFAR19 and MIF decreased obviously,cell apoptosis ratio was significantly increased by TUNEL assay.Characterized morphology was observed by electron microscope(chromatin condensation,nuclear disintegration,formation of apoptotic bodies).Conclusion The protein TFAR19 might enhance apoptosis in PC-3M cells in coordination with MIF and was supposed to be a adjunctive drug of prostate cancer therapy.