RESUMO
OBJECTIVE: To investigate the role of nesprin-1 in mouse embryonic stem cells differentiation into cardiomyocyte. METHODS: Hanging drop-suspension-adherence method was applied for the differentiation of mouse embryonic stem cells into cardiomyocytes under the inducing of salvia miltorrhiza and 5-azacytidine. Changes in nesprin-1 gene expression were detected by using Western blotting and immunofluorescent assay. RNA interference was used to reduce nesprin-1 protein levels to further investigate the importance of nesprin-1 in mouse embryonic stem cells differentiation, group I (target sequence AAAGCCAAGCACGCAACTA), group II (target sequence GGGAACCAACAGTGAGATT), group III (target sequence ACCAGGACATTGCGTACTA), and group IV (control group). RESULTS: The nesprin-1 isoform profile was altered in mouse embryonic stem cells differentiation. The rates of differentiation of the four groups were (17.78 +/- 1.92)%, (36.67 +/- 3.34)%, (44.42 +/- 5.08)%, (77.78 +/- 1.92)%; The rate of differentiation of group IV was higher than RNAi groups and the difference was significant (P < 0.05). In addition, compared with the control group, myosin in RNAi groups were dramatically reduced. CONCLUSION: Nesprin-1 played important roles in mouse embryonic stem cells differentiation into cardiomyocyte. Nesprin-1 isoforms might perform different functions in the process of mouse embryonic stem cells differentiation.
Assuntos
Diferenciação Celular/genética , Células-Tronco Embrionárias/citologia , Miócitos Cardíacos/citologia , Proteínas do Tecido Nervoso/genética , Proteínas Nucleares/genética , Animais , Células Cultivadas , Proteínas do Citoesqueleto , Células-Tronco Embrionárias/metabolismo , Feminino , Masculino , Proteínas de Membrana/genética , Proteínas de Membrana/fisiologia , Camundongos , Proteínas do Tecido Nervoso/fisiologia , Proteínas Nucleares/fisiologia , Interferência de RNARESUMO
LncRNA SBF2-AS1 has been reported to be implicated in the deterioration of multiple human cancers. However, the roles and underlying mechanisms of SBF2-AS1 in acute myeloid leukemia (AML) are still unclear. In the present study, the online GEPIA database showed that SBF2-AS1 expression was significantly increased in AML samples. QRT-PCR results showed that SBF2-AS1 expression was upregulated in AML cells. CCK-8 assay revealed that SBF2-AS1 inhibition decreased AML cells proliferation ability in vitro. Flow cytometry assays showed that SBF2-AS1 inhibition induced AML cells apoptosis and arrested AML cells in G0/G1 phase. Mechanistically, miR-188-5p was identified as a direct target of SBF2-AS1. SBF2-AS1 upregulated the expression level of ZFP91 by sponging miR-188-5p. And the effects of SBF2-AS1 suppression on AML cells progression could be abolished by miR-188-5p inhibitors. Moreover, we found that SBF2-AS1 inhibition reduced tumor growth in vivo. Taken together, our findings elucidated that SBF2-AS1 could act as a miRNA sponge in AML progression, and provided a potential therapeutic strategy for AML treatment.
Assuntos
Leucemia Mieloide Aguda/patologia , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Animais , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Transformação Celular Neoplásica , Feminino , Humanos , Leucemia Mieloide Aguda/genética , Camundongos , Ubiquitina-Proteína Ligases/genética , Regulação para Cima/genéticaRESUMO
AIM: Vitamin D deficiency (VDD) in children is widespread. Therefore, the present study was conducted to assess the prevalence of VDD in schoolchildren from Henan Province, China. METHODS: The study was conducted on school students from areas of Xinxiang, one of the largest cities of Henan Province in China, in the winter of 2013. A questionnaire on demographic data, socioeconomic status, sun exposure, and times of acute respiratory infections (RTI) in the past 12 months were completed for each student. Blood samples were taken to assess serum levels of 25-hydroxyvitamin D [25(OH)D]. RESULTS: In our study, 606 schoolchildren completed the questionnaire. The median serum level of 25(OH)D was 20.5 ng/mL (interquartile range: 15.9-25.1). The prevalence of VDD and insufficiency was 46.9% (284/606) and 85.8% (520/606), respectively. VDD was more associated with female gender, high body mass index, lower socio-professional status, residence in urban areas, and less time spent outdoors. There was an inverse correlation between levels of 25(OH) and times of RTI (p<0.001). CONCLUSION: VDD and insufficiency are common among schoolchildren in Xinxiang. Supplementation with food fortification or vitamin D for Chinese children is warranted.