Downregulation of the c-Fes protein-tyrosine kinase inhibits the proliferation of human renal carcinoma cells
Downregulation of the c-Fes protein-tyrosine kinase inhibits the proliferation of human renal carcinoma cells
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DOI：
10.3892/ijo_00000132
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发表时间：
2009-01-01
影响因子：
5.2
通讯作者：
Smithgall, Thomas E.
中科院分区：
文献类型：
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作者：
Kanda, Shigeru;Miyata, Yasuyoshi;Smithgall, Thomas E.
c-Fes蛋白酪氨酸激酶与造血、神经元、血管内皮和上皮细胞类型的生长和分化相关。在这项研究中，我们研究了小干扰RNA（siRNA）介导的敲低c-Fes表达是否影响人肾癌细胞系ACHN和VMRC-RCW的增殖。免疫荧光显微镜显示c-Fes在这些细胞的胞浆和细胞核中表达，siRNA处理优先下调c-Fes在胞浆中的表达。敲低c-Fes以剂量依赖性方式抑制细胞增殖，细胞死亡增加最小。c-Fes siRNA处理还下调了S473上的AktI和T23上的IKK α的磷酸化以及cyclin D1的表达，增强了I κ B α的表达，并阻止了NF κ B的核定位。用NF κ B抑制肽（SN 50）处理也阻断了这些细胞中NF κ B的增殖和核定位。SN 50处理的效果不被c-Fes siRNA增强，这表明SN 50处理的效果不被c-Fes siRNA增强。下调c-Fes表达通过Akt 1/NF κ B途径抑制细胞周期进程。与siRNA介导的敲除相反，肾癌细胞中野生型或激酶失活的c-Fes的异位表达未能改变其体外和体内的增殖。因此，由siRNA介导的敲低导致的增殖抑制可能取决于c-Fes蛋白的表达而不是其激酶活性。综上所述，我们的研究结果表明下调c-Fes表达可能是一种潜在的治疗方法。
The c-Fes protein-tyrosine kinase is associated with growth and differentiation of hematopoietic, neuronal, vascular endothelial and epithelial cell types. In this study, we investigated whether small interfering RNA (siRNA)mediated knockdown of c-Fes expression affected proliferation of the human renal carcinoma cell lines, ACHN and VMRC-RCW. Immunofluorescence microscopy showed that c-Fes was expressed in both the cytosol and nuclei of these cells, and siRNA treatment preferentially downregulated c-Fes expression in the cytosol. Knock-down of c-Fes inhibited cellular proliferation in a dose-dependent manner with minimal increase in cell death. c-Fes siRNA treatment also downregulated the phosphorylation of AktI on S473 and IKK alpha on T23, and cyclin D1 expression, enhanced the expression of I kappa B alpha, and prevented the nuclear localization of NF kappa B. Treatment with an NF kappa B inhibitory peptide (SN50) also blocked the proliferation and nuclear localization of NF kappa B in these cells. The effect of SN50 treatment was not enhanced by c-Fes siRNA, suggesting that. downregulation of c-Fes expression inhibited cell cycle progression through the Akt1/NF kappa B pathway. In contrast to siRNA-mediated knockdown, ectopic expression of either wild-type or kinase-inactive c-Fes in renal carcinoma cells failed to alter their proliferation in vitro and in vivo. Thus, suppression of proliferation resulting from siRNA-mediated knockdown may depend upon an expression of c-Fes protein rather than its kinase activity. Taken together, our results indicate that downregulation of c-Fes expression may be a potential