ABSTRACT
OBJECTIVE: To investigate the expressions, biological effects and potential mechanism of miR-424 in glioma. METHODS AND METHODS: A total of 54 glioma tissues and 12 normal brain tissues were collected. Human glioma cells (A172, SHG-44, T98, LN18, and LN229) and normal human astrocytes (NHAs) were cultured. Cell invasion and migration capacities were detected by transwell assay. KIF23 was predicted and confirmed as a direct target of miR-424 by TargetScan prediction and Dual-luciferase reporter assay. Six-week-old female nude mice were used for Xenograft tumor formation assay. RESULTS: Results of this study demonstrated a significant decrease of miR-424 expressions both in glioma cells and tissues. Moreover, the declined miR-424 expressions were observed to be correlated with the poor OS and worse clinicopathological parameters of glioma patients. Functional assays indicated that miR-424 restoration could inhibit the glioma cell epithelial-to-mesenchymal transition (EMT) and metastasis, as well as the tumor growth rate and tumor size of glioma mice. Additionally, kinesin family member 23 (KIF23) expressions were found to be significantly enhanced in glioma specimens, and KIF23 was considered to be a functional target for miR-424 in glioma. CONCLUSIONS: MiR-424, considered as a tumor-suppressor, inhibited cell metastasis and EMT by targeting KIF23 in glioma, which may provide a novel insight into tumorigenesis and the basis for the development of miRNA-targeting therapies against glioma.
Subject(s)
Brain Neoplasms/metabolism , Cell Movement , Epithelial-Mesenchymal Transition , Glioma/metabolism , MicroRNAs/metabolism , Microtubule-Associated Proteins/metabolism , Animals , Brain Neoplasms/genetics , Brain Neoplasms/pathology , Cell Line, Tumor , Cell Proliferation , Female , Gene Expression Regulation, Neoplastic , Glioma/genetics , Glioma/secondary , Humans , Male , Mice, Nude , MicroRNAs/genetics , Microtubule-Associated Proteins/genetics , Middle Aged , Neoplasm Invasiveness , Prognosis , Signal Transduction , Tumor BurdenABSTRACT
MiR-200b, a member of the microRNA-200 family, has been identified to be capable of suppressing glioma cell growth through targeting CREB1 or CD133. However, whether miR-200b affects the biological behavior (proliferation, invasion, and migration) of glioma cells is poorly understood. The aim of this study was to evaluate the effect of miR-200b on the biological behavior of glioma cells in vitro. MiRNA-200b mimics, miRNA-200b inhibitor, and mimic control were transfected into conventionally cultured glioma U251 cells, followed by measuring the expression of miR-200b and CD133 in transfected cells by RT-PCR; effect of miR-200b on CD133 mRNA 3'-UTR luciferase activity by luciferase reporter assay; proliferation activity of transfected U251 cells by MTT method; and changes in U251 cell invasion and migration by Transwell method after transfection. Compared to that in the miRNA-200b inhibitor, mimic control, and blank control groups, miRNA-200b expression was significantly increased and CD133 mRNA expression was significantly decreased in the mimic miRNA-200b group in a time-dependent manner (P < 0.05). Meanwhile, dual luciferase reporter assay showed that miR-200b could inhibit CD133 activity through binding to the 3'-UTR of CD133 mRNA (P < 0.05). Furthermore, the proliferation activity and invasion and migration abilities of U251 cells transfected with miRNA-200b mimic were significantly decreased (P < 0.05). In conclusion, overexpression of miR-200b inhibited the proliferation, invasion, and migration of glioma cells possibly through targeting CD133.
Subject(s)
AC133 Antigen/genetics , Glioma/genetics , MicroRNAs/genetics , 3' Untranslated Regions , AC133 Antigen/metabolism , Apoptosis/genetics , Cell Cycle/genetics , Cell Line, Tumor , Cell Movement/genetics , Cell Proliferation/genetics , Gene Expression Regulation, Neoplastic , Glioma/metabolism , Glioma/pathology , Humans , MicroRNAs/biosynthesis , MicroRNAs/metabolism , Nervous System Neoplasms/genetics , Nervous System Neoplasms/metabolism , Nervous System Neoplasms/pathology , RNA, Messenger/genetics , RNA, Messenger/metabolism , TransfectionABSTRACT
BACKGROUND: In the present study thromboelastography (TEG) was to study whether or not hypercoagulopathy might contribute to the thrombosis of implantable central venous access device (Port-A-Cath, Pharmacia) in cancer patients. METHODS: All 76 oncological patients who were enrolled in this study had their R time, alpha angle and MA value measured before Port-A-Cath implantation, of whom 11 patients received re-implantation because of thrombotic device. We compared the measurements of these 11 patients (thrombotic group) with that of 65 patients (control group) who received Port-A-Cath implantation for the first time. According to TEG values the hemostatic status in these patients was classified as hypercoagulable, normal or hypocoagulable for comparison. All patients in the control group were followed up for 3 months for occurrence of thrombosis. RESULTS: It was found that no patient in the thrombotic group was associated with hypercoagulopathy. Five patients (7.5%) in the control group was found in hypercoagulable status at the time of catheter insertion but none of them developed clinical thrombosis during three months of observation. There was no significant difference between the two groups for R time, alpha angle but a higher MA value was found in the control group (p < 0.05). Furthermore, the hypercoagulability (7.5% for the control vs. none for the thrombotic group), hypocoagulability (1.5% vs. 9.1%) and normocoagulability (91.0% vs. 90.9%) were not statistically different between the two groups (Fisher exact test, P = 0.229). CONCLUSIONS: We conclude that hypercoagulopathy in cancer patients has little, if any, contribution in thrombosis of the implantable central venous access device.
