1.Molecular Biology, Affiliated Hospital of LuzhouMedical College from May 2007 to April 2008.
Xinjun LI ; Yingying LIU ; Xiangguo XIA ; Hong XU ; Yi ZENG ; Zengjun ZHOU
Chinese Journal of Tissue Engineering Research 2009;13(19):3776-3780
BACKGROUND: Studies have demonstrated that exogenous neural stem calls (NSCs) could repair nerve and promote recovery of neurofunction following cerebral hemorrhage. However, the influence of internal environment after cerebral hemorrhage on the survival and differentiation of NSCs is a complex and variable process.OBJECTIVE: To observe the survival and differentiation of human embryonic NSCs implanted in rats with cerebra hemorrhage.DESIGN, TIME AND SETTING: Open, immunohistochemistry, experiment was performed at the Laboratory of Molecular Biology, Affiliated Hospital of Luzhou Medical College from May 2007 to April 2008.MATERIALS: A total of 40 female SD rats were provided by the Experimental Animal Institute, Chinese Academy of Medical Sciences. Brain of 8-week aborted fetus was obtained from Department of Gynaecology and Obstetrics, the People's Hospital of Deyang City.METHODS: Cerebral cortex cells of 8-week aborted human fetus were harvested and cultured in vitro to obtain human embryonic NSCs. Cerebral hemorrhage rat models were established via injection of autologous arterial blood in caudate nucleus. Two days after modeling, 5 μL BrdU-labeled human embryonic NSCs suspension was transplanted at four points surrounding hematoma cavity in the rats. After 1 and 2 weeks, rats were sacrificed. Adjacent sections were doubly stained by BrdUImicrotubule-associated protein 2 (MAP-2) and BrdU/glial fibrillary acidic protein (GFAP).MAIN OUTCOME MEASURES: The survival, immigration and differentiation of human embryonic NSOs implanted in rats were observed by immunohistochemistry and immunofluorescance staining.RESULTS: BrdU-positive cells were oval and brown. At 1 and 2 weeks after implantation, BrdU-positive calls survived and migrated, and they migrated more widely at 2 weeks after implantation. At 1 week after implantation, BrdU/MAP-2-positive cells and BrdU/GFAP-positive calls were observed in cerebral tissue sections, and the number of BrdU/MAP-2-positive cells was more than BrdU/GFAP-positive calls; At 2 weeks after implantation, BrdU-positive cells found in choroid plexus and blood capillary were significantly reduced, and BrdU/GFAP-positive calls were more than BrdU/MAP-2- positive cells.CONCLUSION: Implanted human embryonic NSCs can survive and migrate in the hemorrhage region, gradually differentiate into neurons or astrocytes.
2.Regulatory effect of hsa-miR-150-5p on malignant behaviors of glioblastoma U-251MG cells by targeting HIF1α
ZHOU Jiang ; XIA Xiangguo ; CHEN Ligang ; LUO Xin ; AKU Buqian ; GUO Zhangchao
Chinese Journal of Cancer Biotherapy 2018;25(9):878-883
Objective: To explore the effect and possible mechanisms of has-miR-150-5p targeting HIF1α to regulate malignant biological behaviors of glioblastoma (GBM) U-251MG cells. Methods: Real-time quantitative PCR (RT-PCR) was used to detect the expression of miR-150-5p and hypoxia inducible factor 1 (HIF1α) in U-251MG cells. Luciferase report assay was carried out to verify the biological relationship between miR-150-5p and HIF1α and their biological functions in U-251MG cells. The protein expressions of miR150-5pand HIF1α in U-251MG cells were detected by western blotting. The ability of cell migration was detected by wound healing test and cell invasion ability was detected by transwell test. Results: After miR-150-5p mimic transfection, the mRNA expression of HIF1α was significantly reduced in U-251MG cells (P<0.01). Bioinformatics prediction and luciferase reporter assay demonstrated that miR-150-5p down-regulated HIF1α through directly binding to HIF1α 3’-untranslated region (3’-UTR) (all P<0.05). In U-251MG cells, miR-150-5p over-expression significantly inhibited HIF1α expression, cell invasion and migration (all P<0.05). Conclusion: miR150-5p inhibits cell invasion and metastasis through negative regulation of HIF1α, indicating that miR-150-5p and HIF1α were both potential therapeutic targets for glioblastoma.