Construction of antisense RNA expression plasmid for u-PAR and its transfection to highly invasive PC-3M cell subclones
| Autor: | Gong Feili, Li Qingfen, Deng Yao-zu, Ma Ding, Li Zhuoya, Liao Guoning, Feng Youmei |
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| Rok vydání: | 2004 |
| Předmět: |
Male
Cell Biomedical Engineering Receptors Cell Surface Biology Transfection Biochemistry law.invention Receptors Urokinase Plasminogen Activator Biomaterials law Complementary DNA Cell Line Tumor Genetics medicine Humans Neoplasm Invasiveness RNA Antisense Cloning Molecular Earth-Surface Processes Messenger RNA Expression vector Reverse Transcriptase Polymerase Chain Reaction Prostatic Neoplasms Molecular biology Urokinase-Type Plasminogen Activator Antisense RNA Antisense Orientation medicine.anatomical_structure Recombinant DNA Plasmids |
| Zdroj: | Journal of Huazhong University of Science and Technology. Medical sciences = Hua zhong ke ji da xue xue bao. Yi xue Ying De wen ban = Huazhong keji daxue xuebao. Yixue Yingdewen ban. 23(4) |
| ISSN: | 1672-0733 |
| Popis: | To evaluate the specific inhibition of antisense u-PAR on the u-PAR expressions in highly invasive cell subclones and to determine its blocking function in the invasion by those cells, a cDNA fragment of u-PAR obtained by RT-PCR was inserted into a plasmid vector named pcDNA3 in antisense orientation. Then the antisense u-PAR recombinant was transfected into highly invasive cell subclones. The u-PAR expression in neo-resistant cells was examined by RT-PCR and immunohistochemical assay. Compared to the control cells, the content of mRNA and protein of u-PAR in transfected cells decreased sharply, and the rate of inhibition was 53% and 73%, respectively, indicating that an antisense u-PAR might have played a specific inhibitory role in its expression in the cells, which may provide a good cell model for making further investigation of the inhibitory effects of the antisense u-PAR on invasion in highly invasive cell subclones of human prostate carcinoma. |
| Databáze: | OpenAIRE |
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