ABSTRACT
Objective The aim of this study was to investigate the expression of cell cycle checkpoint kinase 1(Chk1)gene in glioblastoma cells( GBM) and its correlation with GBM cell proliferation,tumorigenic activity and prognosis. Methods The ex-pression of Chk1 in GBM cells was selected and analyzed by TCGA database and brain tumor molecular database( Rembrandt),and the level of Chk1 expression in GBM cells was detected by molecular biology techniques such as Western blot and Real-Time PCR. The expression of Chk1 was silenced by siRNA to investigate its effect on proliferation and colony-forming ability of GBM cells. The prognosis survival of GBM patients accompanying with Chk1 expression was analyzed by immunohistochemical staining and Rembrandt database. Results The results of TCGA database and Rembrandt showed that Chk1 gene was highly expressed in GBM tissues. West-ern blot and Real-Time PCR also showed that Chk1 gene was highly expressed in GBM cells. Lentiviral transfection siRNA-specific silencing of Chk1 significantly inhibited proliferation and colony-forming ability of U87 cells( P<0. 01 and P<0. 05). Prognostic survival analysis showed that GBM patients with low expression of Chk1 gene had a significantly better clinical outcome than those of GBM patients with high expression of Chk1 gene(P<0. 001). Conclusion Chk1 gene is overexpressed in GBM cells,up-regula-tion of Chk1 gene expression can promote the growth and proliferation of GBM cells,and Chk1 gene is associated with poor prognosis in GBM patients.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the glycolytic phenotype of SHG44 human glioma cells under hypoxic conditions and the association between cell proliferation and apoptosis and the metabolic status.</p><p><b>METHODS</b>An in vitro hypoxic cell model was established in SHG44 cells using CoCl2. Real-time PCR and Western blotting were used to assess the expressions of hypoxia-inducible factor-1α (HIF-1α) and the enzymes involved in glycolysis including PDK1, PKM2, and LDHA. Intracellular ATP levels were measured by bioluminescence assay to assess the energy metabolic status of SHG44 cells. The viability and apoptosis of the cells were examined using MTT assay and flow cytometry, respectively.</p><p><b>RESULTS</b>The cells in hypoxic culture showed obviously increased expressions of HIF-1α, LDHA, PDK1, and PKM2 at both the mRNA and protein levels as compared to those in normal cell culture. Hypoxia of the cells also resulted in a lowered cell proliferative activity and an increased apoptosis rate with lowered intracellular ATP concentrations and elevated mitochondrial membrane potential.</p><p><b>CONCLUSION</b>Hypoxia can induce a glycolytic phenotype of tumor cells. The sensitivity of tumor cells to hypoxia-induced cell death is directly correlated with their metabolic status.</p>
Subject(s)
Humans , Adenosine Triphosphate , Metabolism , Apoptosis , Carrier Proteins , Metabolism , Cell Hypoxia , Cell Line, Tumor , Cell Proliferation , Central Nervous System Neoplasms , Metabolism , Pathology , Glioma , Metabolism , Pathology , Glycolysis , Hypoxia-Inducible Factor 1, alpha Subunit , Metabolism , Membrane Potential, Mitochondrial , Membrane Proteins , Metabolism , Protein Serine-Threonine Kinases , Metabolism , Thyroid Hormones , MetabolismABSTRACT
Objective To investigate the distribution of nitric oxide synthase (NOS), nerve growth factor receptor (NGFR) and interstitial cells of Caial (ICCs) in Hirschsprung's disease (HD). Methods The distribution of NOS,NGFR and ICCs was studied by using NADPH diaphoruse histochemistry, immanohistochemistry with a monoclonal antibody to human NGFR and the specific polycloual antibody against c-kit in 8 normal controls and 10 cases of HD.Results NOS and NGFR were abundantly present in the myenteric plexus and in the nerve fibers of musculature. ICCs were intensively distributed in the surface of circular musculature and around the myenteric plexus to form a network in normal control colon. In contrast, NOS and NGFR were scarce or absent in the myenteric plexus and in the nerve fibers of musculature, while the hypertrophic nerve trunks were NGFR positive, ICCs were scarcely distributed and the network was disrupted in the aganglionic colon in HD. Conclusion These findings suggest the Involvement of NOS,NGFR and ICCs in the patbophysiology of HD.
ABSTRACT
Objective To study the protective effect of hydrogen sulfide(H2S) on intestinal ischemia-reperfusion(I/R) injury so as to provide a new clinical treatment on I/R.Methods NaHS was taken as a donor of H2S.Forty Sprague-Dawley rats were divided into 5 groups with 8 rats in each group: sham operation group,I/R model group,tetram ethylpyrazine group,and NaHS 7 ?mol/kg and 14 ?mol/kg groups.The contents of superoxidase dismutase(SOD),malondialdehyde(MDA) and glutathione peroxidase(GSH-Px) in serum and intestinal tissue were measured,respectively.The intestinal mucosal injury and histological alteration were observed.Results The content of MDA within serum and intestinal tissue was significantly reduced and the activities of SOD and GSH-Px were significantly increased.H2S could obviously reduce the injury in intestinal mucosa and glands.Conclusion H2S possesses a protective effect on intestinal I/R injury in rats.