摘要
<p><b>OBJECTIVE</b>To establish differential proteomics profiles of glioblastoma cell lines from Chinese, and to provide reference for future basic studies.</p><p><b>METHODS</b>Total protein was extracted from 3 glioblastoma cell lines, CHG-5, TJ899 and TJ905. After normalization, the total protein was presented by two-dimensional (2D) electrophoresis, scanned and analyzed. Some of the identified protein spots were verified by immunocytochemistry of cell lines and immunohistochemistry of solid tumors. The glia cells were used as the control throughout the study.</p><p><b>RESULTS</b>A total of 13 differential protein spots were selected, and eventually 10 were identified as unique proteins. These 10 proteins were involved in cytoskeleton forming, cellular metabolism, tumor migration, stress and inflammatory reaction. Immunocytochemistry and immunohistochemistry further confirmed these proteins present in the solid tumors.</p><p><b>CONCLUSION</b>Distinct differential proteomics profiles exist in glioblastoma cell lines and normal glia cells, likely related to the transformation of normal glia to glioma.</p>
Subject(s)
Humans , Brain Neoplasms , Genetics , Metabolism , Cathepsin D , Metabolism , Cell Line, Tumor , Gene Expression Profiling , Glial Fibrillary Acidic Protein , Metabolism , Glioblastoma , Genetics , Metabolism , Microfilament Proteins , Metabolism , Neuroglia , Metabolism , Proteomics , Methods摘要
<p><b>OBJECTIVE</b>To investigate the effects of endostatin and doxycycline on microcirculation patterns in melanoma and their molecular mechanisms.</p><p><b>METHODS</b>To establish mouse B16 melanoma model by subcutaneous injection of B16 melanoma cell suspension. The mice were divided into 3 experimental groups and 1 control group. To treat the mice in the 3 experimental groups with endostatin, doxycycline, endostatin and doxycycline, respectively, and the control group without any treatment. The tumor volume was measured and recorded to make comparison of their growth rate. To assess the expression of MMP-2, MMP-9 and TIMP-2 by immunohistochemical staining. The three microcirculation patterns of endothelium-dependent vessels, mosaic vessels and vasculogenic mimicry were counted. The activity of MMP-2, MMP-9 between different groups was examined by gelatin zymography.</p><p><b>RESULTS</b>Tumor growth in the three experimental groups was statistically significantly slower than that in the control group. The expression of MMP-2, MMP-9 and TIMP-2 in each treated group was significantly different with that in the control group. The amount of three microcirculation patterns in three experimental groups was less than that of the control group, and the amount of MV and VM in each experimental group was significantly less than that in the control group. By gelatin zymography, the enzyme activity of MMP-9, actived-MMP-2 and MMP-2/proMMP-2 in ES, DOX and ES + DOX group was lower than that in the control group, but the enzyme activity of pro-MMP-2 among the four groups was not significantly different.</p><p><b>CONCLUSION</b>The combined use of doxycycline and endostatin in melanoma can inhibit the expression of MMPs, influencing the formation of different microcirculation patterns in melanoma.</p>
Subject(s)
Animals , Female , Male , Mice , Antineoplastic Agents , Pharmacology , Cell Line, Tumor , Doxycycline , Pharmacology , Drug Combinations , Drug Synergism , Endostatins , Pharmacology , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Melanoma, Experimental , Pathology , Mice, Inbred C57BL , Microcirculation , Microvessels , Pathology , Neoplasm Transplantation , Tissue Inhibitor of Metalloproteinase-2 , Metabolism , Tumor Burden摘要
<p><b>OBJECTIVE</b>To investigate the molecular mechanism of endostatin and doxycycline effect on melanoma growth.</p><p><b>METHODS</b>A B16 melanoma mice model was established by intracutaneous injection of B16 cell suspension. The mice were treated with endostatin, doxycycline, endostatin and doxycycline respectively, the control group received no treatment. A time course study of tumor volume was performed to observe the antitumor effect. The expression of matrix metalloproteinase (MMP-9), MMP-2, TIMP-2 were examined by immunohistochemistry staining.</p><p><b>RESULTS</b>Tumors in endostatin treatment group, doxycycline treatment group, endostatin and doxycycline treatment group grew slower than in the control group. The difference of the average tumor volume in the doxycycline group and control group, in the doxycycline with endostatin treatment group and control group were statistically different. The positive expression ratio of MMP-2, MMP-9, TIMP-2 in each treatment group were statistically different from their control groups (F = 12.79, F = 5.56, F = 4.64; P < 0.05).</p><p><b>CONCLUSION</b>Doxycycline and endostatin are able to inhibit the expression of MMPs and promote expression of TIMP, which ultimately inhibits the growth of B16 melonoma.</p>