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Shi M.-L.,Jiangsu Key Laboratory of Biological Cancer Therapy | Shi M.-L.,Xuzhou Medical College | Mei P.-J.,Xuzhou Medical College | Fan Y.-C.,Xuzhou Medical College | And 4 more authors.
Chinese Journal of Cancer Prevention and Treatment | Year: 2013

OBJECTIVE: To investigate the effect of BRG1 gene in glioma cell migration and invasion and its molecular mechanism. METHODS: Using chemically synthesized small interfering RNA, we transfected BRG1 siRNA into human glioma cell lines U251 and U87 by siLentFect Lipid Reagent. We studied the role of BRG1 in glioma cell migration and invasion by cell migration assay and matrigel invasion assay. We performed western blot to detect TIMP-2 and MMP-2 protein expression. We also detected MMP-2 enzyme activity by gelatin zymography. RESULTS: Compared to the control siRNA treatment, knock down of BRG1 decreased the number of U251 and U87 cells that passed the transwell membrane respectively by 76% (96.38±9.67 vs 23.13±1.20, P=0.0001) and 84% (233.31±19.37 vs 37.33±2.31, P<0.001), which were detected by transwell migration assay. Matrigel invasion assay showed that knock down of BRG1 decreased the number of U251 and U87cells that passed the transwell membrane respectively by 73% (97.30±9.35 vs 26.27±1.05, P=0.0001) and 86% (234.50±15.67 vs 32.83±2.42, P<0.001), which were compared to the control group. Western blotting showed that knock down of BRG1 increased TIMP-2 expression and decreased MMP-2 expression, and gelatin zymography showed that knock down of BRG1 decreased MMP-2 enzyme activity. CONCLUSIONS: Our data indicated that silence of BRG1 in glioma cells inhibited cell migration and invasion abilities. Up-regulation of TIMP-2 expression and down-regulation of MMP-2 expression greatly contributed to the reduced cell invasion and migration abilities. Source


Liu J.-J.,Nanjing Medical University | Liu J.-J.,Jiangsu Key Laboratory of Biological Cancer Therapy | Zhang B.-F.,Jiangsu Key Laboratory of Biological Cancer Therapy | Zhang B.-F.,Xuzhou Medical College | And 11 more authors.
Journal of Immunoassay and Immunochemistry | Year: 2012

RGD peptide (Arg-Gly-Asp tripeptide) binds to integrin V3 and V5, which is selectively expressed in tumor neovasculature and on the surface of some tumor cells. Some studies showed that coupling the RGD peptides to anticancer drugs yielded compounds with increased efficiency against tumors and lowered toxicity to normal tissues. The melanoma differentiation-associated gene-7/interleukin-24 gene (mda-7/IL-24) is a novel tumor-suppressor/cytokine gene that exhibits potent tumor-suppressive activity without damaging normal cells. To enhance the antitumor effect, we inserted a glycine residue into the wild type (mda-7/IL-24) between 164Arg and 165Asp to form a RGD peptide, named RGD-mda-7, then expressed RGD-mda-7 in Escherichia coli. Herein, we describe the expression and purification of RGD-mda-7. We detected the characterizations of immunostimulatory activity, tumor targeting, potent cytopathic effect, and apoptosis inducing exploited by RGD-mda-7 in tumor cells, and also compared these characterizations with wtmda-7/IL-24. The data showed that RGD-mda-7 had more potent tumor targeting and apoptosis-inducing effects than wtmda-7/IL-24. © 2012 Copyright Taylor and Francis Group, LLC. Source

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