ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic recombinant expression vector for retinoblastoma 1 gene (RB-1) and investigate the role of RB-1 in prostate cancer.</p><p><b>METHODS</b>The coding sequence of RB-1 gene tagged with FLAG was amplified from the plasmid CMV-RB by PCR method. The fragment was cloned into CMV expression vector and identified by restriction enzyme digestion and sequence analysis. Western Blotting was used to detect RB-1 expression and immunofluorescence was used to observe RB-1 distribution in PC-3 cells transfected with the recombinant.</p><p><b>RESULTS</b>The expression vector CMV-FLAG-RB was successfully constructed as confirmed by PCR, endonuclease digestion and DNA sequence analysis. RB-1 protein was highly expressed and showed a nuclear distribution in PC-3 cells transfected with the recombinant.</p><p><b>CONCLUSIONS</b>The eukaryotic expression vector for RB-1 has been successfully constructed and can be efficiently expressed in PC-3 cells. The expression of RB-1 is located in the cell nuclei.</p>
Subject(s)
Humans , Male , Base Sequence , Cell Line, Tumor , Cloning, Molecular , Gene Expression , Genetic Vectors , Genetics , Molecular Sequence Data , Prostatic Neoplasms , Pathology , Recombinant Proteins , Genetics , Retinoblastoma Protein , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To study the differential expressions of the receptor for advanced glycation end products (RAGE) in the tissues of prostate cancer and normal prostate, and to find the role of RAGE in the pathogenesis of prostate cancer.</p><p><b>METHODS</b>We collected the tissue of prostate cancer and that of normal prostate from the same patient, and compared the differential expressions of RAGE at the tissue, protein and mRNA levels between prostate cancer and normal prostate tissues of 10 patients by immunohistochemistry, Western blot and real-time quantitative PCR.</p><p><b>RESULTS</b>Immunohistochemistry exhibited a significantly higher expression of RAGE in the prostate cancer tissue than in the normal prostate tissue; Western blot showed that the RAGE protein expression was 2.13 times higher in the former than in the latter (P < 0.05); and real-time quantitative PCR revealed the RAGE mRNA expression of the former to be 4.2 times that of the latter (P < 0.05).</p><p><b>CONCLUSION</b>RAGE may play an important role in the pathogenesis and progression of prostate cancer.</p>