【摘要】 目的 研究沉默CT45A1(cancer/testis antigen family 45 member A1)基因的表达对人前列腺癌细胞的体外增殖、迁移和侵袭能力的影响，并初步探讨分子作用机制。方法 实时荧光定量PCR和蛋白免疫印迹(Western blot, WB)检测CT45A1在具有不同侵袭力的人前列腺癌细胞中的表达水平，利用慢病毒感染方式构建CT45A1基因沉默的前列腺癌细胞，MTT法、划痕愈合实验和Transwell小室法评估沉默CT45A1基因对细胞的增殖、迁移和侵袭能力的影响。实时荧光定量PCR和WB检测上皮-间质转化(epithelial-mesenchymal transition, EMT)的重要标志物E-钙黏蛋白(E-cadherin)和波形蛋白(Vimentin)以及关键调控因子Twist和Zeb-1的表达情况。结果 荧光定量PCR和WB实验结果表明，人前列腺癌PC-3细胞中CT45A1 mRNA的相对表达量(4.37±0.34)和CT45A1的蛋白相对表达水平(2.49±0.12),均明显高于LNCap细胞；shCT45A1#1、shCT45A1#2细胞中的CT45A1 mRNA和蛋白表达水平均低于shScram细胞，差异均有统计学意义(P<0.05)。CT45A1基因沉默后，PC-3细胞的增殖水平未发生明显变化(P>0.05),但迁移与侵袭能力及Vimentin、Zeb-1的表达均下降，E-cadherin的表达上调，差异均有统计学意义(P<0.05)。结论 CT45A1可以通过促进Zeb-1介导的EMT来增强PC-3细胞的迁移和侵袭能力。更多还原

【Abstract】 Objective To investigate the impact of express of Cancer/Testis Antigen Family 45 Member A1(CT45A1) on the in vitro proliferative, migratory and invasive capacities of prostate cancer cells, and to dissect its functional mechanism. Methods The levels of CT45A1 expression in human prostate cancer cells with varying invasive capacity were measured using real-time fluorescent quantitative polymerase chain reaction and Western blot analyses. Lentiviral transduction was used to generate prostate cancer cells with CT45A1 knockdown. The effect of CT45A1 knockdown on proliferation, migration, and invasion was assessed using the MTT method, wound healing assay, and Transwell assay, respectively. The levels of E-cadherin and vimentin were measured as markers of epithelial-mesenchymal transition(EMT), along with two transcription factors that regulate EMT, Twist and Zeb-1. The measurements were taken using real-time fluorescent quantitative polymerase chain reaction and WB analysis. Results The results of fluorescence quantitative PCR and WB experiments indicate that the relative expression of CT45A1 mRNA(4.37±0.34) and the protein expression level of CT45A1 in PC-3 cells of human prostate cancer were significantly higher than that in LNCap cells. Additionally, the CT45A1 mRNA and protein expression levels in shCT45A1#1 and shCT45A1#2 cells were lower than those in shScram cells, and the differences were all statistically significant(P<0.05). After silencing the CT45A1 gene, the proliferation level of PC-3 cells did not change significantly(P>0.05). However, the migration and invasion abilities were decreased, and the expression of Vimentin and Zeb-1 were also decreased, while the expression of E-cadherin was up-regulated. All of these differences were statistically significant(P<0.05). Conclusions CT45A1 may augment the motility and invasiveness of prostate cancer PC-3 cells in vitro by triggering EMT mediated by Zeb-1.更多还原