Zhu Wenjing, Hu Xiaohua, Xu Jiguo, Cheng Yi, Shao Yiqun, Peng Yu
Department of Urology, Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, 200437, China.
Cell Biochem Biophys. 2015 May;72(1):171-7. doi: 10.1007/s12013-014-0433-3.
We sought to study the effects of PI3K/Akt pathway and its downstream substrate NF-κB on prostate cancer bone metastatic process. Expression level of active p-Akt in PC3 cells was upregulated by transient expression with constitutively active plasmid CA-Akt or, alternatively, suppressed by dominant negative construct DN-Akt. NF-κB activity was determined by luciferase reporter assays. mRNA and protein expressions of receptor activator of NF-κB ligand (RANKL), parathyroid hormone-related protein (PTHrP), and bone morphogenetic protein 2 (BMP-2) were evaluated using RT-PCR and Western blotting. The effect of cross-talk between PC3 and SaOS2 cells on cell proliferation was analyzed using a co-culture system. Stimulation of p-Akt promoted NF-κB activity, and led to an increase in mRNA and protein expressions of RANKL, PTHrP, and BMP-2 in PC3 PCa cells through NF-κB. Co-culturing PC3 and SaOS2 cells significantly increased the expression of p-Akt and the activity of NF-κB, and promoted proliferation of both PC3 and SaOS2 cells. Increasing expression levels of p-Akt by transfection with CA-Akt led to further increase in cells proliferation, whereas NF-κB inhibitor PDTC partially blocked this effect. PI3K/Akt pathway stimulates the expressions of RANKL, PTHrP, and BMP-2 partly through NF-κB, suggesting its importance for bone metastasis of prostate carcinoma. Interaction of prostate cancer cells with bone cells has a stimulatory effect on cell proliferation.
我们试图研究PI3K/Akt信号通路及其下游底物NF-κB对前列腺癌骨转移过程的影响。通过用组成型活性质粒CA-Akt瞬时转染上调PC3细胞中活性p-Akt的表达水平，或者用显性负性构建体DN-Akt抑制其表达。通过荧光素酶报告基因测定法测定NF-κB活性。使用RT-PCR和蛋白质印迹法评估NF-κB配体受体激活剂（RANKL）、甲状旁腺激素相关蛋白（PTHrP）和骨形态发生蛋白2（BMP-2）的mRNA和蛋白质表达。使用共培养系统分析PC3和SaOS2细胞之间的相互作用对细胞增殖的影响。p-Akt的激活促进了NF-κB活性，并通过NF-κB导致PC3前列腺癌细胞中RANKL、PTHrP和BMP-2的mRNA和蛋白质表达增加。PC3和SaOS2细胞共培养显著增加了p-Akt的表达和NF-κB的活性，并促进了PC3和SaOS2细胞的增殖。用CA-Akt转染增加p-Akt的表达水平导致细胞增殖进一步增加，而NF-κB抑制剂PDTC部分阻断了这种作用。PI3K/Akt信号通路部分通过NF-κB刺激RANKL、PTHrP和BMP-2的表达，表明其对前列腺癌骨转移的重要性。前列腺癌细胞与骨细胞的相互作用对细胞增殖有刺激作用。
