臭椿酮对前列腺癌22RV1细胞内源性AR等的影响

Effect of ailanthone on endogenous androgen receptor and its downstream genes PSA,FKBP5,SLC45A3,NDRG1 mRNA expression in prostate cancer 22RV1 cells

目的探究臭椿酮对前列腺癌22RV1细胞内源性雄激素受体(androgen receptor,AR)及其下游基因前列腺特异性抗原(prostate-specific antigen,PSA)、FK506结合蛋白5(FK506 binding protein 5,FKBP5)、溶质转运蛋白家族45A3(solute carriers 45A3,SLC45A3)、N-myc下游调节基因(N-myc downstream regulated gene 1,NDRG1)mRNA表达的影响。方法取处于对数生长期的前列腺癌22RV1细胞,随机分为4组,分别为对照组、臭椿酮低剂量组、臭椿酮中剂量组和臭椿酮高剂量组,每组设置5个复孔。臭椿酮低、中和高剂量组分别采用0.4、0.8和1.0μmol·L^(-1)臭椿酮干预,对照组加入等量磷酸盐缓冲液(phosphate buffer saline,PBS)干预。用二甲基噻唑(dimethylthiazole,MTT)实验检测干预24、48、72 h后细胞的增殖情况;用流式细胞仪检测干预72 h后细胞的凋亡率;用实时荧光定量PCR检测干预72 h后各组内源性AR及其下游基因PSA、FKBP5、SLC45A3、NDRG1 mRNA的相对表达量。结果臭椿酮低、中和高剂量组MTT实验吸光度(A)及AR、PSA、FKBP5、SLC45A3 mRNA的相对表达量均低于对照组,臭椿酮高剂量组低于臭椿酮低、中剂量组,臭椿酮中剂量组MTT实验A值低于臭椿酮低剂量组(P<0.05);臭椿酮低、中和高剂量组的凋亡率、NDRG1 mRNA的相对表达量均高于对照组,臭椿酮高剂量组高于臭椿酮低、中剂量组,臭椿酮中剂量组高于臭椿酮低剂量组(P<0.05)。结论臭椿酮可抑制前列腺癌22RV1细胞的增殖,促进其凋亡,其中1.0μmol·L^(-1)臭椿酮的作用最强,可能与其调控内源性AR及其下游基因PSA、FKBP5、SLC45A3和NDRG1 mRNA的表达有关。

Objective To investigate the effect of ailanthone on endogenous androgen receptor(AR)and its downstream genes prostate-specific antigen(PSA),FK506 binding protein 5(FKBP5),solute carriers 45A3(SLC45A3),N-myc downstream regulated gene 1(NDRG1)mRNA expression in prostate cancer 22RV1 cells.Methods Prostate cancer 22RV1 cells were randomly divided into 4 groups,the control group,the ailanthone low-dose,medium-dose and high-dose groups,5 compound holes in each group.The ailanthone low,medium and high dose groups were treated with 0.4,0.8 and 1.0μmol·L^(-1) ailanthone,respectively.The control group were treated with an equal amount of phosphate buffer saline(PBS).The cell proliferation was detected by dimethylthiazole(MTT)test at 24,48,and 72 hours after intervention.The cell apoptotic rate was detected by flow cytometry at 72 hours after intervention.The endogenous AR and its downstream genes PSA,FKBP5,SLC45A3,NDRG1 mRNA expression were detected by real-time PCR.Results The MTT test optical density(A)value and the endogenous AR and its downstream genes PSA,FKBP5,SLC45A3 mRNA expression of 3 ailanthone dose groups were lower than those of the control group.The ailanthone high-dose group were lower than those of the ailanthone low and medium dose groups.The ailanthone medium-dose group were lower than those of the ailanthone low-dose group(P<0.05).The apoptotic rate and the NDRG1 mRNA expression of ailanthone 3 dose group were higher than those of control group.Meanwhile the ailanthone highdose group were higher than those of the ailanthone low and medium dose groups,and the ailanthone medium-dose group were higher than those of the ailanthone low-dose group(P<0.05).Conclusion Ailanthone can inhibit the proliferation and promote the apoptosis of prostate cancer 22RV1 cells.1.0μmol·L^(-1) ailanthone had the strongest effect.The mechanism may be related to its regulation of endogenous AR and downstream genes PSA,FKBP5,SLC45A3 and NDRG1 mRNA expression.