RESUMO
ZMYND11 (zinc finger MYND-type containing 11) has been widely regarded to be involved in a variety of cancers as a potential suppressor. However, the biological role and mechanism of ZMYND11 in glioblastoma multiform (GBM) remain unknown. In this study, we found that ZMYND11 expression was remarkably decreased in GBM tissues from 20 cases and cell line (U87) compared to normal brain tissue from 10 cases (P < 0.001). Furthermore, we explored that ZMYND11 upregulation significantly suppressed U87 cells proliferation and invasion, induced cell cycle arrest and apoptosis in vitro. Subsequently, we identified increased ZMYND11 inhibited the tumor growth using tumor cells xenograft experiment on rude mice. Moreover, we explored that ZMYND11 was a new direct and functional target of miR-196a-5p in U87 via luciferase reporter assay. In addition, we confirmed the negative correlation between miR-196a-5p and ZMYND11 in GBM tissue and U87 cells by changing the expression level of miR-196a-5p with lentivirus and plasmid vector. Furthermore, we demonstrated that decreased ZMYND11 could reverse suppressive effect of downregulated miR-196a-5p on U87 by rescue experiment. Taken together, ZMYND11 was demonstrated to be a potential and extremely promising suppressor of GBM, while miRNA-196a-5p was quite an important target of treatment of GBM.
Assuntos
Neoplasias Encefálicas/metabolismo , Neoplasias Encefálicas/patologia , Proteínas de Transporte/metabolismo , Glioblastoma/metabolismo , Glioblastoma/patologia , MicroRNAs/metabolismo , Adulto , Idoso , Animais , Proteínas de Ciclo Celular , Linhagem Celular Tumoral , Proliferação de Células , Proteínas Correpressoras , Proteínas de Ligação a DNA , Regulação para Baixo , Feminino , Humanos , Masculino , Camundongos , Camundongos Nus , Pessoa de Meia-Idade , Invasividade NeoplásicaRESUMO
Gliomas are the most common and aggressive types of tumors in human brain, of which the prognosis remains dismal because of their biological behavior. The involvement of miRNAs in tumorigenesis of various kinds of cancers drives us to explore new miRNAs related to gliomas. We measured expression level of miR953p by qRT-PCR in human glioma and non-neoplasm brain tissues and found that higher level of miR953p in glioma tissues of higher grade. Biological functions of miR953p on glioma cells were investigated by MTT assay, flow cytometry and transwell assay. We discovered the cell lines transfected with miR953p ASO (antisense oligonucleotide) had retarded proliferation and invasion but enhanced apoptosis ability. We searched on-line tool Targetscan and selected CELF (CUGBP- and ETR-3-like family 2) as a putative target. Luciferase reporter was employed to confirm the binding sites in 3'UTR region of CELF2 for miR953p. The correlation between expression of CELF2 and miR953p was determined by western blotting and qRT-PCR both in cell lines and human samples. Results showed CELF2 was a direct target of miR953p and expression levels of CELF2 and miR953p were negatively correlated. Finally, CELF2 largely abrogated the effects of miR953p on proliferation, invasion and apoptosis of glioma cells in rescue experiments, which verified the role of CELF2 in miR953p regulating glioma biological behavior. In conclusion, our data suggest the expression level of miR953p is positively related to glioma grade and downregulation of miR953p affects proliferation, invasion and apoptosis of glioma cells by targeting CELF2. We identified miR953p as a putative therapeutic target and CELF2 as a potential tumor suppressor.