Glioblastoma multiforme (GBM) is the most aggressive form of brain tumor. Despite radical surgery and radiotherapy supported by chemotherapy, the disease still remains incurable with an extremely low median survival rate of 12-15 months from the time of initial diagnosis. The main cause of treatment failure is considered to be the presence of cells that are resistant to the treatment. MicroRNAs (miRNAs) as regulators of gene expression are involved in the tumor pathogenesis, including GBM. MiR-338 is a brain-specific miRNA which has been described to target pathways involved in proliferation and differentiation. In our study, miR-338-3p and miR-338-5p were differentially expressed in GBM tissue in comparison to non-tumor brain tissue. Overexpression of miR-338-3p with miRNA mimic did not show any changes in proliferation rates in GBM cell lines (A172, T98G, U87MG). On the other hand, pre-miR-338-5p notably decreased proliferation and caused cell cycle arrest. Since radiation is currently the main treatment modality in GBM, we combined overexpression of pre-miR-338-5p with radiation, which led to significantly decreased cell proliferation, increased cell cycle arrest, and apoptosis in comparison to irradiation-only cells. To better elucidate the mechanism of action, we performed gene expression profiling analysis that revealed targets of miR-338-5p being Ndfip1, Rheb, and ppp2R5a. These genes have been described to be involved in DNA damage response, proliferation, and cell cycle regulation. To our knowledge, this is the first study to describe the role of miR-338-5p in GBM and its potential to improve the sensitivity of GBM to radiation.
- MeSH
- buněčné dělení účinky léků účinky záření MeSH
- glioblastom genetika patologie MeSH
- kontrolní body buněčného cyklu účinky léků účinky záření MeSH
- lidé středního věku MeSH
- lidé MeSH
- membránové proteiny biosyntéza genetika MeSH
- mikro RNA genetika MeSH
- monomerní proteiny vázající GTP biosyntéza genetika MeSH
- nádorové buněčné linie MeSH
- nádorové proteiny biosyntéza genetika MeSH
- nádory mozku genetika patologie MeSH
- neuropeptidy biosyntéza genetika MeSH
- poškození DNA genetika MeSH
- proteinfosfatasa 2 biosyntéza genetika MeSH
- regulace genové exprese u nádorů genetika MeSH
- RNA nádorová genetika MeSH
- stanovení celkové genové exprese MeSH
- tolerance záření genetika MeSH
- transportní proteiny biosyntéza genetika MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Expression of doublecortin (DCX), a 43-53 kDa microtubule binding protein, is frequently used as (i) an early neuronal marker to identify the stage of neuronal maturation of in vivo grafted neuronal precursors (NSCs), and (ii) a neuronal fate marker transiently expressed by immature neurons during development. Reliable identification of the origin of DCX-immunoreactive cells (i.e., host vs. graft) requires detailed spatial and temporal mapping of endogenous DCX expression at graft-targeted brain or spinal cord regions. Accordingly, in the present study, we analyzed (i) the time course of DCX expression in pre- and postnatal rat and porcine spinal cord, and (ii) the DCX expression in spinally grafted porcine-induced pluripotent stem cells (iPS)-derived NSCs and human embryonic stem cell (ES)-derived NSCs. In addition, complementary temporospatial GFAP expression study in porcine spinal cord was also performed. In 21-day-old rat fetuses, an intense DCX immunoreactivity distributed between the dorsal horn (DH) and ventral horn was seen and was still present in the DH neurons on postnatal day 20. In animals older than 8 weeks, no DCX immunoreactivity was seen at any spinal cord laminae. In contrast to rat, in porcine spinal cord (gestational period 113-114 days), DCX was only expressed during the pre-natal period (up to 100 days) but was no longer present in newborn piglets or in adult animals. Immunohistochemical analysis was confirmed with a comparable expression profile by western blot analysis. Contrary, the expression of porcine GFAP started within 70-80 days of the pre-natal period. Spinally grafted porcine iPS-NSCs and human ES-NSCs showed clear DCX expression at 3-4 weeks postgrafting. These data indicate that in spinal grafting studies which employ postnatal or adult porcine models, the expression of DCX can be used as a reliable marker of grafted neurons. In contrast, if grafted neurons are to be analyzed during the first 4 postnatal weeks in the rat spinal cord, additional markers or grafted cell-specific labeling techniques need to be employed to reliably identify grafted early postmitotic neurons and to differentiate the DCX expression from the neurons of the host.
- MeSH
- časové faktory MeSH
- druhová specificita MeSH
- embryonální kmenové buňky metabolismus transplantace MeSH
- indukované pluripotentní kmenové buňky metabolismus transplantace MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- lidé MeSH
- mícha růst a vývoj metabolismus MeSH
- neurogeneze fyziologie MeSH
- neuropeptidy biosyntéza MeSH
- potkani Wistar MeSH
- prasata MeSH
- proteiny asociované s mikrotubuly biosyntéza MeSH
- těhotenství MeSH
- transplantace kmenových buněk trendy MeSH
- vývojová regulace genové exprese * MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Klíčová slova
- hypokretiny, orexin-A (hypokretin-1), orexin-B (hypokretin-2),
- MeSH
- energetický metabolismus fyziologie MeSH
- lidé MeSH
- narkolepsie * etiologie MeSH
- neuropeptidy * biosyntéza fyziologie sekrece MeSH
- obezita etiologie MeSH
- orexinové receptory MeSH
- regulace chuti k jídlu * fyziologie MeSH
- stravovací zvyklosti fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- hipokampus MeSH
- hormon uvolňující gonadotropiny MeSH
- hormon uvolňující thyreotropin MeSH
- hormony kůry nadledvin metabolismus škodlivé účinky MeSH
- nemoci centrálního nervového systému patofyziologie MeSH
- neuroendokrinologie * MeSH
- neuropeptidy biosyntéza farmakokinetika MeSH
- neuropsychiatrie MeSH
- neurotransmiterové látky MeSH
- psychický stres metabolismus patofyziologie MeSH
- systém hypofýza - nadledviny * fyziologie patofyziologie sekrece účinky léků MeSH
