【摘要】 目的：探讨姜黄素对人前列腺癌C4-2细胞和LNCaP细胞增殖、迁移及侵袭的影响，并阐明其可能的作用机制。方法：采用慢病毒转染系统分别转染C4-2细胞和LNCaP细胞，作为shCD147-C4-2组和shCD147-LNCaP组。采用RNA干扰技术制备沉默CD147基因细胞，以转入空载体的细胞作为阴性对照，分为shNC-C4-2组（shNC-C4-2细胞）和shNC-LNCaP组（shNC-LNCaP细胞）。取生长对数期C4-2、LNCap、shCD147-C4-2和shCD147-LNCaP细胞，加入20μmol·L^-1姜黄素，处理0和24 h时，显微镜观察各组细胞形态表现。噻唑蓝（MTT）法检测各组细胞增殖活性，细胞划痕实验检测各组细胞迁移率，Western blotting法检测各组细胞中凋亡、侵袭和迁移相关蛋白表达水平。结果：与C4-2组比较，沉默CD147基因后shCD147-C4-2组细胞中CD147蛋白表达量明显减少；与LNCaP组比较，沉默CD147基因后shCD147-LNCaP组细胞中CD147蛋白表达量明显减少。与处理0 h比较，20μmol·L^-1姜黄素处理24 h后C4-2组和LNCaP组部分细胞出现凋亡征象，且有典型凋亡小体存在；shCD147-C4-2组和shCD147-LNCaP组细胞凋亡现象减弱。MTT法检测，与C4-2+0μmol·L^-1姜黄素组比较，C4-2+20μmol·L^-1姜黄素组、C4-2+40μmol·L^-1姜黄素组、C4-2+60μmol·L^-1姜黄素组和C4-2+80μmol·L^-1姜黄素组细胞增殖活性均明显降低（P<0.01）；与LNCaP+0μmol·L^-1姜黄素组比较，LNCaP+20μmol·L^-1姜黄素组、LNCaP+40μmol·L^-1姜黄素组、LNCaP+60μmol·L^-1姜黄素组和LNCaP+80μmol·L^-1姜黄素组细胞增殖活性均明显降低（P<0.01）；与shNC-C4-2组比较，shNC-C4-2+20μmol·L^-1姜黄素组细胞增殖活性明显降低（P<0.01）；与shNC-C4-2+20μmol·L^-1姜黄素组比较，shCD147-C4-2+20μmol·L^-1姜黄素组细胞增殖活性明显升高（P<0.01）；与shNC-LNCaP组比较，shNC-LNCaP+20μmol·L^-1姜黄素组细胞增殖活性明显降低（P<0.01）；与shNC-LNCaP+20μmol·L^-1姜黄素组比较，shCD147-LNCaP+20μmol·L^-1姜黄素组细胞增殖活性明显升高（P<0.01）。细胞划痕愈合实验检测，姜黄素处理24 h，与C4-2组比较，C4-2+20μmol·L^-1姜黄素组和C4-2+40μmol·L^-1姜黄素组细胞迁移率均明显降低（P<0.01）；与LNCaP组比较，LNCaP+20μmol·L^-1姜黄素组和LNCaP+40μmol·L^-1姜黄素组细胞迁移率均明显降低（P<0.01）；与shNC-C4-2组比较，shNC-C4-2+20μmol·L^-1姜黄素组细胞迁移率明显降低（P<0.01）；与shNC-C4-2+20μmol·L^-1姜黄素组比较，shCD147-C4-2+20μmol·L^-1姜黄素组细胞迁移率明显升高（P<0.05）；与shNC-LNCaP组比较，shNC-LNCaP+20μmol·L^-1姜黄素组细胞迁移率明显降低（P<0.01）；与shNC-LNCaP+20μmol·L^-1姜黄素组比较，shCD147-LNCaP+20μmol·L^-1姜黄素组细胞迁移率明显升高（P<0.05）。Western blotting法检测，与C4-2组比较，C4-2+20μmol·L^-1姜黄素组和C4-2+40μmol·L^-1姜黄素组细胞中B细胞淋巴瘤2 （Bcl-2）相关X蛋白（Bax）、裂解的含半胱氨酸的天冬氨酸蛋白酶3 （cleaved Caspase-3）和聚二磷酸腺苷（ADP）-核糖聚合酶1 （PARP1）蛋白表达水平均明显升高（P<0.01）,Bcl-2蛋白表达水平均明显降低（P<0.05或P<0.01）；与LNCaP组比较，LNCaP+20μmol·L^-1姜黄素组和LNCaP+40μmol·L^-1姜黄素组细胞中Bax、cleaved Caspase-3和PARP1蛋白表达水平均明显升高（P<0.01）,LNCaP+40μmol·L^-1姜黄素组Bcl-2蛋白表达水平明显降低（P<0.01）；与shNC-C4-2组比较，shNC-C4-2+20μmol·L^-1姜黄素组细胞中Bax、cleaved Caspase-3和PARP1蛋白表达水平均明显升高（P<0.01）,Bcl-2蛋白表达水平明显降低（P<0.05）；与shNC-C4-2+20μmol·L^-1姜黄素组比较，shCD147-C4-2+20μmol·L^-1姜黄素组细胞中Bax和cleavedCaspase-3蛋白表达水平均明显降低（P<0.01）。与shNC-LNCaP组比较，shNCLNCaP+20μmol·L^-1姜黄素组细胞中Bax、 cleaved Caspase-3和PARP1蛋白表达水平均明显升高（P<0.05或P<0.01）,Bcl-2蛋白表达水平明显降低（P<0.05）；与shNC-LNCaP+20μmol·L^-1姜黄素组比较，shCD147-LNCaP+20μmol·L^-1姜黄素组细胞中Bax、cleaved Caspase-3和PARP1蛋白表达水平均明显降低（P<0.05或P<0.01）,Bcl-2蛋白表达水平明显升高（P<0.05）。与C4-2组比较，C4-2+20μmol·L^-1姜黄素组和C4-2+40μmol·L^-1姜黄素组细胞中E-钙黏蛋白（E-cadherin）蛋白表达水平均明显升高（P<0.01），神经钙黏蛋白（N-cadherin）和波形蛋白（Vimentin）蛋白表达水平均明显降低（P<0.01）；与LNCaP组比较，LNCaP+20μmol·L^-1姜黄素组和LNCaP+40μmol·L^-1姜黄素组细胞中E-cadherin蛋白表达水平均明显升高（P<0.01）,LNCaP+40μmol·L^-1姜黄素组细胞中N-cadherin和Vimentin蛋白表达水平均明显降低（P<0.01）；与shNC-C4-2组比较，shNC-C4-2+20μmol·L^-1姜黄素组细胞中N-cadherin和Vimentin蛋白表达水平均明显降低（P<0.01）；与shNCC4-2+20μmol·L^-1姜黄素组比较，shCD147-C4-2+20μmol·L^-1姜黄素组细胞中E-cadherin蛋白表达水平明显降低（P<0.01）,N-cadherin和Vimentin蛋白表达水平均明显升高（P<0.01）；与shNC-LNCaP组比较，shNC-LNCaP+20μmol·L^-1姜黄素组细胞中E-cadherin蛋白表达水平明显升高（P<0.01）,N-cadherin和Vimentin蛋白表达水平均明显降低（P<0.01）；与shNC-LNCaP+20μmol·L^-1姜黄素组比较，shCD147-LNCaP+20μmol·L^-1姜黄素组细胞中E-cadherin蛋白表达水平明显降低（P<0.01）,N-cadherin表达水平明显升高（P<0.05）。结论：姜黄素对体外前列腺癌细胞增殖、迁移和侵袭有抑制作用，并诱导细胞凋亡，沉默CD147基因可在一定程度上降低其抑制作用和诱导细胞凋亡能力。更多还原

【Abstract】 Objective:To discuss the effect of curcumin on the proliferation, migration, and invasion of the human prostate cancer C4-2 and LNCaP cells, and to clarify its possible mechanism. Methods:The lentiviral transfection system was used to transfect the C4-2 and LNCaP cells, regarded as shCD147-C4-2group and shCD147-LNCaP group. RNA interference technology was used to prepare the CD147-silenced cells; the cells transfected with an empty vector were regarded as negative control and divided into shNC-C4-2 group（shNC-C4-2 cells） and shNC-LNCaP group（shNC-LNCaP cells）. The C4-2 and LNCaP cells at logarithmic growth phase, as well as shCD147-C4-2 and shCD147-LNCaP cells, were treated with 20 μmol·L^-1 curcumin. The morphology of the cells in various groups was observed under microscope at 0 and 24 h of treatment; MTT method was used to detect the proliferation activities of the cells in various groups; cell scratch assay was used to detect the migration rates of the cells in various groups;Western blotting method was used to detect the expression levels of apoptosis, invasion, and migration-related proteins in the cells in various groups. Results:Compared with C4-2 group, the expression of CD147 protein in the cells in shCD147-C4-2 group was significantly decreased after CD147gene silenting. Compared with LNCaP group, the expression level of CD147 protein in the cells in shCD147-LNCaP group was significantly decreased after CD147 gene silenting. Compared with 0 h of treatment, some cells in C4-2 and LNCaP groups after 24 h of treatment with 20 μmol·L^-1 curcumin, showed apoptosis signs with the presence of typical apoptotic bodies. The apoptotic phenomena in shCD147-C4-2 and shCD147-LNCaP groups was reduced. The MTT assay results showed that compared with C4-2+0 μmol·L^-1 curcumin group, the proliferation activities of the cells in C4-2+20 μmol·L^-1curcumin group, C4-2+40 μmol·L^-1 curcumin group, C4-2+60 μmol·L^-1 curcumin group, and C4-2+80 μmol·L^-1 curcumin group were decreased（P<0. 01）. Compared with LNCaP+0 μmol·L^-1 curcumin group, the proliferation activity of the cells in LNCaP+20 μ mol·L^-1 curcumin group, LNCaP+40 μmol·L^-1 curcumin group, LNCaP+60 μmol·L^-1 curcumin group, and LNCaP+80 μmol·L^-1 curcumin group were decreased（P<0. 01）. Compared with shNC-C4-2 group, the proliferation activity of the cells in shNC-C4-2+20 μmol·L^-1 curcumin group was decreased（P<0. 01）. Compared with shNC-C4-2+20 μmol·L^-1 curcumin group, the proliferation activity of the cells in shCD147-C4-2+20 μmol·L^-1curcumin group was increased（P<0. 01）. Compared with shNC-LNCaP group, the proliferation activity of the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group was decreased（P<0. 01）; compared with shNC-LNCaP+20 μmol·L^-1 curcumin group, the proliferation activity of the cells in shCD147-LNCaP+20 μmol·L^-1 curcumin group was significantly increased（P<0. 01）. The cell scratch healing assay results showed that compared with C4-2 group, the migration rates of the cells in C4-2+20 μmol·L^-1 curcumin group and C4-2+40 μ mol·L^-1 curcumin group after 24 h of treatment were decreased（P<0. 01）; compared with LNCaP group, the migration rates of the cells in LNCaP+20 μmol·L^-1 curcumin group and LNCaP+40 μmol·L^-1 curcumin group were increased（P<0. 01）; compared with shNC-C4-2 group, the migration rate of the cells in shNC-C4-2+20 μmol·L^-1 curcumin group was decreased（P<0. 01）; compared with shNC-C4-2+20 μmol·L^-1 curcumin group, the migration rate of the cells in shCD147-C4-2+20 μmol·L^-1 curcumin group was significantly increased（P<0. 05）; compared with shNC-LNCaP group, the migration rate of the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group was decreased（P<0. 01）; compared with shNC-LNCaP+20 μmol·L^-1 curcumin group, the garation rate of the cells in shCD147-LNCaP+20 μmol·L^-1 curcumin group was significantly increased（P<0. 05）. The Western blotting results showed that compared with C4-2 group, the expression levels of Bcl-2-associated X protein（Bax）, cleaved Caspase-3, and poly ADP-ribose polymerase 1（PARP1） proteins in the cells in C4-2+20 μmol·L^-1 curcumin group and C4-2+40 μmol·L^-1 curcumin group were significantly increased（P<0. 01）, and the expression levels of Bcl-2 protein was significantly decreased（P<0. 05 or P<0. 01）; compared with LNCaP group, the expression levels of Bax, cleaved Caspase-3, and PARP1 proteins in the cells in LNCaP+20 μmol·L^-1 curcumin group and LNCaP+40 μmol·L^-1 curcumin group were significantly increased（P<0. 01）, and the expression level of Bcl-2 protein in the cells in LNCaP+40 μmol·L^-1curcumin group was decreased（P<0. 01）; compared with shNC-C4-2 group, the expression levels of Bax, cleaved Caspase-3, and PARP1 proteins in the cells in shNC-C4-2+20 μmol·L^-1 curcumin group were significantly increased（P<0. 05 or P<0. 01）, and the expression level of Bcl-2 protein was significantly decreased（P<0. 05）; compared with shNC-C4-2+20 μmol·L^-1 curcumin group, the expression levels of Bax and cleaved Caspase-3 proteins in the cells in shCD147-C4-2+20 μmol·L^-1curcumin group were significantly decreased（P<0. 01）; compared with shNC-LNCaP group, the expression levels of Bax, cleaved Caspase-3, and PARP1 proteins in the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group were significantly increased（P<0. 05 or P<0. 01）, and the expression level of Bcl-2 protein was significantly decreased（P<0. 05）; compared with shNC-LNCaP+20 μmol·L^-1curcumin group, the expression levels of Bax, cleaved Caspase-3, and PARP1 proteins in the cells in shCD147-LNCaP+20 μmol·L^-1 curcumin group were significantly decreased（P<0. 05 or P<0. 01）, and the expression level of Bcl-2 protein was significantly increased（P<0. 05）. Compared with C4-2 group, the expression levels of E-cadherin protein in the cells in C4-2+20 μmol·L^-1 curcumin group and C4-2+40 μ mol·L^-1 curcumin group were significantly increased（P<0. 01）, and the expression levels of N-cadherin and Vimentin proteins were significantly decreased（P<0. 01）; compared with LNCaP group, the expression levels of E-cadherin protein in the cells in LNCaP+20 μmol·L^-1 curcumin group and LNCaP+40 μmol·L^-1 curcumin group were significantly increased（P<0. 01）, and the expression levels of N-cadherin and Vimentin proteins in the cells in LNCaP+40 μmol·L^-1 curcumin group were significantly decreased（P<0. 01）; compared with shNC-C4-2 group, the expression levels of N-cadherin and Vimentin proteins in the cells in shNC-C4-2+20 μmol·L^-1 curcumin group were significantly decreased（P<0. 01）; compared with shNC-C4-2+20 μ mol·L^-1 curcumin group, the expression level of E-cadherin protein in the cells in shCD147-C4-2+20 μmol·L^-1 curcumin group was significantly decreased（P<0. 01）, and the expression levels of N-cadherin and Vimentin proteins were significantly increased（P<0. 01）; compared with shNC-LNCaP group, the expression level of E-cadherin protein in the cells in shNC-LNCaP+20 μmol·L^-1 curcumin group was significantly increased（P<0. 01）, and the expression levels of N-cadherin and Vimentin proteins were significantly decreased（P<0. 01）; compared with shNC-LNCaP+20 μmol·L^-1 curcumin group, the expression level of E-cadherin protein in the cells in shCD147-LNCaP+20 μ mol·L^-1 curcumin group was significantly decreased（P<0. 01）, and the expression level of N-cadherin was significantly increased（P<0. 05）. Conclusion:Curcumin inhibits the proliferation, migration, and invasion of the prostate cancer cells in vitro and induces the apoptosis; silencing the CD147 gene partially reduces its inhibitory effect and its ability to induce the apoptosis.更多还原