前列腺癌PC-3细胞抗TRAIL诱导凋亡的实验研究

Experimental Study of Anti-TRAIL-induced Apoptosis in Prostate Cancer PC-3 Cells

目的肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apoptosis inducing ligand,TRAIL)能够诱导人前列腺癌细胞株PC-3细胞凋亡而不损伤正常细胞。部分PC-3细胞能够对TRAIL产生抵抗作用,本研究对其发生发展的分子机制进行进一步探讨和阐述。方法将体外培养的PC-3细胞分为对照组(仅加培养基)和实验组(不同浓度滴度的TRAIL处理),作用时间24~72小时。MTT法检测细胞抑制率,筛选出抵抗TRAIL治疗的PC-3细胞,Q-PCR检测Fas相关死亡域样白介素1B转换酶抑制蛋白(FLICE inhibitory protein,FLIP)、死亡受体4(death receptor,DR4)、DR5、Fas相关死亡域(Fas-associated death domain,FADD)和caspase-8基因表达情况。结果 TRAIL对PC-3细胞具有一定增殖抑制作用,细胞平均存活率随着TRAIL作用时间延长而减少;同时与TRAIL浓度有关,在200ng/ml的TRAIL时,对PC-3细胞的增殖抑制作用较强。Q-PCR检测结果显示TRAIL作用24小时的PC-3细胞较对照组的FLIP、DR4、DR5、FADD和caspase-8基因的表达降低;随着TRAIL作用时间延长至72小时,DR4、DR5、FADD和caspase-8基因的表达下降,而FLIP表达上升。结论体外实验TRAIL对于PC-3治疗具有一定效果,和TRAIL浓度作用时间有关,在TRAIL作用一定时间内,随着作用时间延长,细胞增殖抑制作用越强。TRAIL对PC-3抑制作用与药物浓度有一定关系,在200ng/ml时对PC-3细胞抑制生长率最强。PC3-TR的DR4、DR5、FADD和caspase-8及FLIP在TRAIL短时间治疗内表达下降,而在长时间治疗时DR4、DR5、FADD和caspase-8表达下降,FLIP表达上升。

Objective Tumor necrosis factor - related apoptosis inducing ligand （ TRAIL）promotes apoptosis in PC - 3 cells while sparing normal cells. Although many PC - 3 cells are sensitive to TRAIL - induced apoptosis,some tumor cells could evade the proapoptotic effects of TRAIL. The molecular mechanism of TRAIL.resistant tumors is not fully clear,so it is necessary to discuss the relevant mechanism in this paper. MethodsThe PC - 3 cells were divided into control group （ only with medium）and experimental groups （ processed withdifferent concentrations of TRAIL for 24 - 72 hours in vitro. Cell inhibition rate was determined by MTT methodand then TRAIL - resistant PC - 3 tumors cells were selected. The expression levels of FLICE inhibitory protein（ FLIP） ,death receptor 4 （ DR4） ,DR5,Fas - associated death domain （ FADD）and caspase - 8 genes between TRAIL - sensitive and TRAIL - resistant tumor cells were detected by Q - PCR. Results TRAIL could inhibit the proliferation of PC - 3 cell with a certain extent,and the cell average inhibition rate was enhanced duration of action TRAIL;Meanwhile,inhibition rate was also associated with TRAIL concentration. With the strongest cell growth inhibition rate at 200 ng / ml TRAIL. The Q - PCR detecting expression levels of FLIP,DR4,DR5,FADD and caspase - 8 gene in TRAIL - resistance PC - 3 cell were lower than those in the TRAIL - sensitive PC - 3 cell within 24 hours TRAIL effect. As the duration of TRAIL,expression levels of FLIP,DR5,FADD and caspase - 8 gene in TRAIL - resistance PC - 3 cell were lower than those in TRAIL - sensitive PC -3 cell,but the expression levels of FLIP was higher than that in TRAIL - sensitive PC - 3 cell within 72 hours.Conclusions TRAIL has a certain therapeutic effect for PC - 3 in vitro,which has a certain relationship the concentration and therapy time of TRAIL. TRAIL has a certain relationship the concentration of TRAIL,with thestrongest cell growth inhibition rate at 200ng / ml TRAIL. The express levels of DR4,DR5,FADD,caspase - 8and FLIP in PC3 cell decreased with a 24 hours treatment of TRAIL;the express levels of DR4,DR5,FADDand caspase - 8 decreased and FLIP increased in PC3 cell with a 72 hours treatment of TRAIL.