Knockdown of Long Noncoding RNA LUCAT1 Inhibits Cell Viability and Invasion by Regulating miR-375 in Glioma.

Recently, long noncoding RNAs (lncRNAs) have emerged as new gene regulators and prognostic markers in several cancers, including glioma. Here we focused on lncRNA LUCAT1 on the progression of glioma. qRT-PCR was used to determine the expression of LUCAT1 and miR-375 in glioma tissues and cells. MTT and Transwell invasion assays were performed to determine the function of LUCAT1 in glioma progression. The bioinformatics tool DIANA was used to predict the targets of LUCAT1. Pearson's correlation analysis was performed to explore the correlation between LUCAT1 and miR-375. In the present study, we showed that LUCAT1 was substantially upregulated in glioma tissues and cells. LUCAT1 inhibition significantly suppressed the proliferation and invasion of glioma cells. Subsequently, DIANA showed that miR-375 was predicted to contain the complementary binding sites to LUCAT1. Luciferase reporter assay showed that miR-375 directly targeted LUCAT1. In addition, we found that miR-375 was downregulated in glioma tissues and negatively correlated with LUCAT1 expression in glioma tissues. Furthermore, the results showed that miR-375 could rescue the function of LUCAT1 in glioma progression. The lncRNA LUCAT1 was critical for the proliferation and invasion of glioma cells by regulating miR-375. Our findings indicated that LUCAT1 might offer a potential novel therapeutic target for the treatment of glioma.

Overexpression of miR­21 promotes neural stem cell proliferation and neural differentiation via the Wnt/ß­catenin signaling pathway in vitro.

The primary aim of the present study was to examine the effects of microRNA­21 (miR­21) on the proliferation and differentiation of rat primary neural stem cells (NSCs) in vitro. miR­21 was overexpressed in NSCs by transfection with a miR­21 mimic. The effects of miR­21 overexpression on NSC proliferation were revealed by Cell Counting kit 8 and 5­ethynyl­2'­deoxyuridine incorporation assay, and miR­21 overexpression was revealed to increase NSC proliferation. miR­21 overexpression was confirmed using reverse transcription­quantitative polymerase chain reaction (RT­qPCR). mRNA and protein expression levels of key molecules (ß­catenin, cyclin D1, p21 and miR­21) in the Wnt/ß­catenin signaling pathway were studied by RT­qPCR and western blot analysis. RT­qPCR and western blot analyses revealed that miR­21 overexpression increased ß­catenin and cyclin D1 expression, and decreased p21 expression. These results suggested that miR­21­induced increase in proliferation was mediated by activation of the Wnt/ß­catenin signaling pathway, since overexpression of miR­21 increased ß­catenin and cyclin D1 expression and reduced p21 expression. Furthermore, inhibition of the Wnt/ß­catenin pathway with FH535 attenuated the influence of miR­21 overexpression on NSC proliferation, indicating that the factors activated by miR­21 overexpression were inhibited by FH535 treatment. Furthermore, overexpression of miR­21 enhanced the differentiation of NSCs into neurons and inhibited their differentiation into astrocytes. The present study indicated that in primary rat NSCs, overexpression of miR­21 may promote proliferation and differentiation into neurons via the Wnt/ß­catenin signaling pathway in vitro.

Neural Stem Cell-based Intraocular Administration of Pigment Epithelium-derived Factor Promotes Retinal Ganglion Cell Survival and Axon Regeneration after Optic Nerve Crush Injury in Rat: An Experimental Study.

BACKGROUND: Pigment epithelium-derived factor (PEDF) is regarded as a multifunctional protein possessing neurotrophic and neuroprotective properties. PEDF has a very short half-life, and it would require multiple injections to maintain a therapeutically relevant level without a delivery system. However, multiple injections are prone to cause local damage or infection. To overcome this, we chose a cell-based system that provided sustained delivery of PEDF and compared the effect of weekly injections of PEDF and neural stem cell (NSC)-based intraocular administration of PEDF on retinal ganglion cell (RGC) survival and axon regeneration after optic nerve injury. METHODS: Seventy-two rats were randomly assigned to 3 groups: group with injections of phosphate buffered saline (PBS) (n=24), group with weekly injections of PEDF (n=24), and group with NSC-based administration of PEDF (n=24). Western blot was used to analyze the PEDF protein level 2 weeks after injection. Retinal flat mounts and immunohistochemistry were employed to analyze RGC survival and axon regeneration 2 weeks and 4 weeks after injection. The data were analyzed with one-way ANOVA in SPSS (version 19.0). A P<0.05 was considered significant. RESULTS: The PEDF protein level in the group with NSC-based administration of PEDF increased compared with that in the groups with injections of PEDF and PBS (P<0.05). The PEDF-modified NSCs differentiated into GFAP-positive astrocytes andß-tubulin-III-positive neurons. NSC-based administration of PEDF effectively increased RGC survival and improved the axon regeneration of the optic nerve compared with weekly injections of PEDF. CONCLUSION: Subretinal space transplantation of PEDF-secreting NSCs sustained high concentrations of PEDF, differentiated into neurons and astrocytes, and significantly promoted RGC survival and axon regeneration after optic nerve injury.