ABSTRACT
Objective The long non-coding RNA (lncRNA) MTHFD2 gene is expressed differentially in glioblastoma (GBM) and normal brain tissues, but its biological role in tumors, and particularly in GBM, remains unclear. This study aims to investigate the expression of lncRNA MTHFD2 in the GBM tissue and four GBM cell lines, and explore the effect of its down-regulated expression on the biological function of GBM cells. Methods Specimens of GBM and the paracancerous tissue (as normal control) were collected from 9 patients treated by surgical resection in our Department of Neurosurgery between September and December 2017 LV-MTHFD2-shRNA (U251 shRNA and U-87MG shRNA groups) and empty LV-control solution (U251 shRNA and U-87MG control groups) were transfected into the U251 and U-87MG cell lines. The expressions of lncRNA MTHFD2 in the GBM tissue and the GBM cell lines were detected by qRT-PCR, the chemosensitivity and proliferation of the cells after transfection measured by CCK-8 assay, and the changes in the cell migration ability determined by Transwell assay. Results The relative expression of lncRNA MTHFD2 was significantly higher in the GBM than in the normal tissue (5.13 ± 3.96 vs 1.27 ± 0.58, P < 0.05), while that of MTHFD2 was remarkably lower in the U251 shRNA than in the U251 control group (0.05 ± 0.01 vs 1.00 ± 0.00, P < 0.01), and so was that in the U-87MG shRNA than in the U-87MG control (P < 0.05). The number of cells penetrating the Transwell membrane was markedly lower in the U251 shRNA group than in the U251 control (41.4 ± 6.99 vs 125.8 ± 25.27 per field of view, P < 0.01), and so was that in the U-87MG shRNA than in the U-87MG control (P < 0.05). CCK-8 assay showed that, at 4 days after transfection, the A value was significantly decreased in the U251 shRNA and U-87MG shRNA groups as compared with the U251 control and U-87MG control groups (P < 0.05). Cellular drug resistance test manifested remarkably reduced fifty percent inhibitory concentrations (IC50) in the U251 shRNA and U-87MG shRNA groups as compared with the U251 control and U-87MG control groups (P < 0.05). Conclusion DDown-regulation of the expression of lncRNA MTHFD2 can inhibit the proliferation and migration of U251 and U-87MG cells and enhance the chemosensitivity of the cells to temozolomide, which suggests that lncRNA MTHFD2 could be a potential therapeutic target against GBM.
ABSTRACT
<p><b>UNLABELLED</b>OBJECTIVE To study the potential interaction between PrP protein.</p><p><b>METHODS</b>The supernatant of health and scrapie-infected hamsters' brain homogenate was prepared, while various recombinant 14-3-3beta or PrP proteins were purified. The possible molecular interaction between 14-3-3beta proteins and PrP was tested by pull-down and immunoprecipitation assays.</p><p><b>RESULTS</b>Both native PrP(c) and its protease-resistant isoform (PrP(Sc)) formed complexes with 14-3-3beta. The full-length recombinant 14-3-3beta proteins interacted with PrP. The domain responsible for interacting 14-3-3beta was located at N-terminal of 14-3-3beta (residues 1 to 38).</p><p><b>CONCLUSION</b>The studies of the association of PrP with 14-3-3beta may further provide insight into a potential role of 14-3-3beta in the biological function of PrP and the pathogenesis of prion disease.</p>
Subject(s)
Animals , Cricetinae , 14-3-3 Proteins , Metabolism , Binding Sites , Brain Chemistry , Endopeptidases , Metabolism , PrPSc Proteins , Metabolism , Prion Diseases , Pathology , Prions , Metabolism , ScrapieABSTRACT
<p><b>OBJECTIVE</b>To explore the effects of aberrant expression of sperm protein 17 (Sp17) on the migration of the ovarian cancer cell line HO-8910.</p><p><b>METHODS</b>The recombinant plasmid pEGFP-Sp17 containing Sp17 and enhanced green fluorescent protein gene was transfected into the human ovarian cancer cell line HO-8910 with Lipofectamine 2000. The expression of Sp17 was examined by RT-PCR and Western blot, and the cell migratory capability detected by Transwell chamber assays.</p><p><b>RESULTS</b>Sp17 was expressed as a fusion protein with EGFP after transfected. There was a significant difference in the migratory cell number of the transfected and the control cells (156.6 +/- 14.9/HP vs 39.3 +/- 8.53/HP, P < 0.05).</p><p><b>CONCLUSION</b>The aberrant expression of Sp17 greatly enhances the migration of ovarian cancer cells.</p>