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建立稳定表达外源PC-1基因的人前列腺癌骨转移C4-2B细胞模型,初步探讨PC-1基因表达对前列腺癌发展的影响.通过脂质体介导的方法,将融合PC-1基因的真核表达载体pcDNA3·1-PC-1稳定转染C4-2B细胞,Western印迹和RT-PCR技术,分别从蛋白水平和RNA水平确定外源PC-1基因表达.MTT和软琼脂集落形成能力等一系列方法,研究PC-1基因的功能,RT-PCR和实时定量PCR检测前列腺癌发生发展相关基因表达的变化.结果表明,PC-1基因的高表达能够诱导雄激素受体(AR)调控基因和一系列重要的信号通路成员基因PSA、PSMA、NKX3·1、Jagged1、EphA3、SGEF和NOTCH3等表达发生变化.实验结果初步证明,PC-1基因表达在晚期前列腺癌中,以及在雄激素非依赖的转变中可以发挥作用,PC-1基因表达可调控一些重要信号通路.对PC-1基因功能深入研究将有可能为发现新的前列腺癌的诊断治疗分子靶标提供线索.
To establish a cell model of bone metastasis of human prostate cancer C4-2B stably expressing exogenous PC-1 gene and to explore the effect of PC-1 gene expression on the development of prostate cancer.Through liposome-mediated method, PcDNA3.1-PC-1 was transfected into C4-2B cells stably and the expression of exogenous PC-1 gene was determined by protein level and RNA level respectively.MTT and soft agar colony formation Ability, and so on, and to study the function of PC-1 gene.The changes of gene expression related to the development of prostate cancer were detected by RT-PCR and real-time quantitative PCR.The results showed that high expression of PC-1 gene could induce the expression of androgen receptor ) Regulatory genes and a series of important signaling pathway members such as PSA, PSMA, NKX3 · 1, Jagged1, EphA3, SGEF and NOTCH3 etc. The results of the preliminary experiments show that PC-1 gene expression in advanced prostate cancer, Androgen-independent transition can play a role, PC-1 gene expression can regulate some important signaling pathways.Depth of PC-1 gene function may provide clues for the discovery of new molecular targets for the diagnosis and treatment of prostate cancer.