AMACR在前列腺癌中的表达及沉默AMACR基因对前列腺癌DU145细胞增殖凋亡的影响  

The expression of AMACR in prostate cancer and the effect of silencing AMACR gene on the proliferation and apoptosis of prostate cancer DU145 cells

作　　者：李华华 岳姣姣[2] 牛虹 唐静雯 岳光星 LI Huahua;YUE Jiaojiao;NIU Hong;TANG Jingwen;YUE Guangxing(Department of Integrated Traditional Chinese and Western Medicine,Affiliated Cancer Hospital of Zhengzhou University(He’nan Cancer Hospital),Zhengzhou 450003,He’nan,China;Department of Encephalopathy,Third Affiliated Hospital of He’nan University of Traditional Chinese Medicine,Zhengzhou 450003,He’nan,China)

机构地区：[1]郑州大学附属肿瘤医院(河南省肿瘤医院)中西医结合科,郑州450003 [2]河南中医药大学第三附属医院脑病科,郑州4500030

出　　处：《癌症进展》2020年第3期252-257,共6页Oncology Progress

摘　　要：目的探讨α-甲基酰基辅酶A消旋酶(AMACR)在前列腺癌中的表达及沉默AMACR基因对前列腺癌DU145细胞增殖凋亡的作用机制。方法取55例前列腺癌患者的前列腺癌组织及32例良性前列腺增生症患者的良性前列腺增生组织。免疫组化检测两种组织中AMACR表达情况并分析与前列腺癌患者临床特征的关系。将前列腺癌细胞株DU145分为Blank组、pcDNA-AMACR组、pcDNA-AMACR NC组、siRNA-AMACR组、siRNA-AMACR NC组,分别转染相应质粒。实时荧光定量聚合酶链反应(qRT-PCR)和Western blot法分别检测细胞中AMACR、核因子-kappa B抑制蛋白(IκBα)、核因子κB(NF-κB)、Ki-67、B细胞淋巴瘤/白血病-2(Bcl-2)、B细胞淋巴瘤-2相关X蛋白(BAX)蛋白和mRNA的表达情况,CCK-8法检测各组细胞增殖情况,流式细胞仪检测各组细胞凋亡情况。结果前列腺癌组织中AMACR的阳性表达率明显高于良性前列腺增生组织(P﹤0.01)。AMACR表达与前列腺癌患者TNM分期、淋巴结转移、病理分级、肿瘤分期和术前前列腺特异性抗原(PSA)水平有关(P﹤0.05)。与pcDNA-AMACR NC组相比,pcDNA-AMACR组AMACR、IκBα、NF-κB、Ki-67、Bcl-2蛋白和mRNA表达水平均上升,BAX蛋白和mRNA表达水平均下降,细胞增殖能力上升,细胞凋亡率降低。同时,与siRNA-AMACR NC组相比,siRNA-AMACR组AMACR、IκBα、NF-κB、Ki-67、Bcl-2蛋白和mRNA表达水平均降低,BAX蛋白和mRNA表达水平均上升,细胞增殖能力降低,细胞凋亡率升高。结论下调AMACR基因会抑制NF-κB信号通路的激活,从而抑制前列腺癌细胞的增殖并诱导凋亡。Objective To investigate the expression of alpha-methylacyl-CoA racemase(AMACR)in prostate cancer and the mechanism of silencing AMACR gene on the proliferation and apoptosis of prostate cancer DU145 cells.Method Fifty-five patients with prostate cancer and 32 cases with benign prostatic hyperplasia were included in the analysis,from them the prostate cancer tissues and benign prostatic hyperplasia tissues were collected,respectively.Immunohistochemistry was used to detect the expression of AMACR in these specimens,and to investigate the relationship between AMACR expression and clinical features of prostate cancer patients.Prostate cancer cells DU145 were divided into five groups:Blank group,pcDNA-AMACR group,pcDNA-AMACR NC group,siRNA-AMACR group and siRNA-AMACR NC group,and each was transfected with corresponding plasmids.The expression of AMACR,inhibitor of nuclear factor kappa B(IκBα),nuclear factor kappa B(NF-κB),Ki-67,B cell lymphoma/leukemia-2(Bcl-2),Bcl-2-associated X protein(BAX)mRNA and protein were detected by quantitative real-time polymerase chain reaction(qRT-PCR)and Western blot,respectively.Meanwhile,CCK-8 assay was used to detect cell proliferation and flow cytometry was used to detect cell apoptosis.Result The positive expression of AMACR was significantly higher in prostate cancer tissues versus benign prostate hyperplasia tissues(P<0.01).AMACR expression was related to TNM stage,lymph node metastasis,pathological grade,tumor stage and preoperative prostate specific antigen(PSA)level(P<0.05).Compared with pcDNA-AMACR NC group,the expression levels of AMACR,IκBα,NF-κB,Ki-67 and Bcl-2 protein and mRNA in pcDNA-AMACR group increased significantly,while the expression levels of BAX protein and mRNA decreased,besides,higher proliferative ability and lower apoptosis rate were observed.Compared with siRNA-AMACR NC group,the expression levels of AMACR,IκBα,NF-κB,Ki-67 and Bcl-2 protein and mRNA in siRNA-AMACR group decreased significantly,while the expression levels of BAX protein and mRNA in

关 键 词：AMACR基因 NF-ΚB信号通路 前列腺癌细胞 增殖 凋亡 

分 类 号：R737.25[医药卫生—肿瘤]