Mohan P M, Lakka S S, Mohanam S, Kin Y, Sawaya R, Kyritsis A P, Nicolson G L, Rao J S
Department of Neurosurgery, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Clin Exp Metastasis. 1999;17(7):617-21. doi: 10.1023/a:1006779902978.
We previously showed that downregulation of the urokinase-type plasminogen activator receptor (uPAR) in the SNB19 human glioblastoma cell line by the stable transfection of a plasmid expressing a 300 bp antisense sequence to the 5' end of the uPAR gene produced a decrease in the amount of target mRNA. In a more recent study, we found that adenovirus-mediated transduction (Ad-uPAR) of the same uPAR antisense gene construct in SNB19 cells also downregulated uPAR protein levels. We report here that Ad-uPAR-transfected SNB19 cells produced the same amounts of target uPAR mRNA but significantly less protein by in vitro translation and by in situ [35S] labeling compared to Ad-CMV vector-transfected and mock-transfected cells. This antisense construct also inhibited glioblastoma cell invasion confirming previous results. We conclude that downregulation of uPAR by this antisense gene construct results from inhibition of protein translation.
我们之前表明，通过稳定转染一个表达针对尿激酶型纤溶酶原激活物受体（uPAR）基因5'端300 bp反义序列的质粒，在SNB19人胶质母细胞瘤细胞系中下调uPAR，会使靶mRNA的量减少。在最近的一项研究中，我们发现，在SNB19细胞中腺病毒介导的相同uPAR反义基因构建体转导（Ad-uPAR）也下调了uPAR蛋白水平。我们在此报告，与Ad-CMV载体转染和mock转染的细胞相比，Ad-uPAR转染的SNB19细胞产生的靶uPAR mRNA量相同，但通过体外翻译和原位[35S]标记产生的蛋白量显著减少。这种反义构建体也抑制了胶质母细胞瘤细胞的侵袭，证实了先前的结果。我们得出结论，这种反义基因构建体对uPAR的下调是由于蛋白质翻译受到抑制。
