MicroRNA-16 inhibits glioma cell growth and invasion through suppression of BCL2 and the nuclear factor-κB1/MMP9 signaling pathway.
Bottom Line: MicroRNA-16 decreased glioma malignancy by downregulating NF-κB1 and MMP9, and led to suppressed invasiveness of human glioma cell lines SHG44, U87, and U373.Our results also indicated that upregulation of miR-16 promoted apoptosis by suppressing BCL2 expression.Finally, the upregulation of miR-16 in a nude mice model of human glioma resulted in significant suppression of glioma growth and invasiveness.
Affiliation: Department of Neurosurgery and Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, Suzhou, China.Show MeSH
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Mentions: The microRNA.org database (http://www.microrna.org/microrna/get Mrna.do?gene=596&utr=5412&organism=9606) shows that BCL2 is a target of post-transcriptional repression by miR-16 (Fig.3a). Our data showed that miR-16 expression was inversely correlated with BCL2 in the same tissue samples of human glioma (Fig.3b). A dual-luciferase reporter assay system was used to validate whether miR-16 directly recognizes the 3′-UTR region of BCL2 mRNA. As shown in Figure3(c), the relative luciferase activities of the pre-miR-16 groups transfected with BCL2 3′-UTR constructs were significantly decreased, compared to those transfected with pMIR-BCL2 constructs, for the 293T cell line, implying that BCL2 is a direct target of miR-16. To examine if miR-16 facilitates apoptosis in human glioma cells, flow cytometry data indicated that increasing miR-16 expression induced early apoptosis, compared with the negative control oligonucleotide and the mock, and that the proportion of early apoptotic cells in the miR-16 treatment group was markedly increased (P < 0.01, n = 3) (Fig. S1). We further investigated the mechanism of transfection of the miR-16 mimics and inhibitors on inducing apoptosis in human glioma cells. BCL2 was detected by Western blot. Data showed that the upregulation of miR-16 led to obvious downregulation of BCL2 expression (Fig. 3d), and that miR-16 induces apoptosis in human glioma cells in vitro.
Affiliation: Department of Neurosurgery and Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, Suzhou, China.