Institution of Biochemistry and Molecular Biology, Medical School of Shandong University, Jinan, China.
Mol Biol Rep. 2010 Mar;37(3):1505-12. doi: 10.1007/s11033-009-9549-8. Epub 2009 May 22.
NKX3.1, a prostate-specific gene, plays an important role in prostate development and carcinogenesis. However, its precise function has not been established. In present study, we transfected the NKX3.1 eukaryotic expression plasmid (pcDNA3.1-NKX3.1) into human prostate cancer cells PC-3, which lack of NKX3.1 expression, and established stable transfectants. Then, we investigated the influence of NKX3.1 on the cell growth, cell migration and colony formation efficiency. The results showed that restoration of NKX3.1 expression inhibited proliferation and invasion activities of PC-3 cells. Further, a cDNA microarray containing 22,000 human genes was used to identify the gene expression differences. The results showed that there were 1,953 genes showing more than a two-fold difference in expression. Subsequent ontological analysis revealed that a large proportion of the classified genes were related to cell growth, cell signal and cell invasion. Finally, the expression of Caspase-3, Bcl-2, P27, Cdk6 and AMACR, randomly selected genes from microarray data, was validated by RT-PCR and western blot. Collectively, our results first analyzed the gene expression profile in PC-3 cells induced by NKX3.1 and indicated that NKX3.1 might exert its function by regulating the expression of relative genes.
NKX3.1 是一种前列腺特异性基因,在前列腺发育和癌变中发挥着重要作用。然而,其确切功能尚未确定。在本研究中,我们将 NKX3.1 真核表达质粒(pcDNA3.1-NKX3.1)转染至缺乏 NKX3.1 表达的人前列腺癌细胞 PC-3 中,建立稳定转染细胞系。然后,我们研究了 NKX3.1 对细胞生长、细胞迁移和集落形成效率的影响。结果表明,恢复 NKX3.1 表达抑制了 PC-3 细胞的增殖和侵袭活性。进一步,使用包含 22000 个人类基因的 cDNA 微阵列鉴定基因表达差异。结果表明,有 1953 个基因的表达差异超过两倍。随后的本体分析显示,分类基因的很大一部分与细胞生长、细胞信号和细胞侵袭有关。最后,通过 RT-PCR 和 Western blot 验证了从微阵列数据中随机选择的 Caspase-3、Bcl-2、P27、Cdk6 和 AMACR 等基因的表达。综上所述,我们的研究结果首次分析了 NKX3.1 诱导的 PC-3 细胞中的基因表达谱,表明 NKX3.1 可能通过调节相关基因的表达发挥其功能。