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MicroRNA-34a induces apoptosis in the human glioma cell line,A172, through enhanced ROS production and NOX2 expression
Authors:San-Zhong Li  Yi-Yang Hu  Jing Zhao  Yong-Bo Zhao  Ji-Dong Sun  Yue-fan Yang  Chen-Cheng Ji  Zao-Bin Liu  Wei-Dong Cao  Yan Qu  Wei-Ping Liu  Guang Cheng  Zhou Fei
Affiliation:1. Department of Neurosurgery, Xi-jing Hospital, Fourth Military Medical University, Xi’an 710032, China;2. State Key Laboratory of Cancer Biology, Department of Medical Genetics and Developmental Biology, Fourth Military Medical University, Xi’an 710032, China;3. Department of Anesthesiology, Xi-jing Hospital, Fourth Military Medical University, Xi’an 710032, China
Abstract:

Background

MicroRNA is a type of non-coding small RNA involved in regulating genes and signaling pathways through incomplete complementation with target genes. Recent research supports key roles of miRNA in the formation and development of human glioma.

Methods

The relative quantity of miR-34a was initially determined in human glioma A172 cells and glioma tissues. Next, we analyzed the impact of miR-34a on A172 cell viability with the MTT assay. The effects of miR-34a overexpression on apoptosis were confirmed with flow cytometry and Hoechst staining experiments. We further defined the target genes of miR-34a using immunofluorescence and Western blot.

Results

MiR-34a expression was significantly reduced in human glioma A172 cells and glioma tissue, compared with normal glial cells and tissue samples. Our MTT data suggest that up-regulation of miR-34a inhibits cell viability while suppression of miR-34a enhances cell viability. Flow cytometry and Hoechst staining results revealed increased rates of apoptosis in A172 human glioma cells overexpressing miR-34a. Using immunofluorescence and Western blot analyses, we identified NOX2 as a target of miR-34a in A172 cells.

Conclusion

MiR-34a serves as a tumor suppressor in human glioma mainly by decreasing NOX2 expression.
Keywords:miR-34a   Human glioma cell line A172   Apoptosis   NOX2
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