The Function Of Zfx Gene In Human Brain Gliomas

The human brain glioma is the most common malignant intracranial tumor. In recent years the treatment don’t breakthrough, without significantly improve of the prognosis in the disease, so we need to explore the new treatment approach.Today gene therapy is considered one of the most application prospect of treatment means. Choose the right target genes is the key step in gene therapy.The X chromosome coupling zinc finger protein (zinc finger protein, X-linked, Zfx) is a member of the zinc finger protein family.Zfx gene is a common genetic basis of self-renewal in embryonic and adult stem cells.But the report of the expression and function of Zfx gene in malignant glioma is less at home and abroad.So we study the detection of the expression of Zfx gene in malignant glioma by the PCR technology and study the function of Zfx in malignant glioma U251cells by transfecting the lentivirus-meiated small interfering RNAs (siRNAs) into U251.This study contains two parts of work:the first, to analyze the expression of Zfx mRNA in human glioma tissues and glioma cell lines.The second, to used lentivirus-mediated small interfering RNAs (siRNAs) to knock down Zfx expression in the human malignant glioma cell line U251, to assessed the effects of Zfx knockdown on cell proliferation, cell cycle and apoptosis.Part Ⅰ:The expression of the Zfx gene in human gliomaObjective To analyze the expression of Zfx mRNA in human glioma tissues and in SHG44cells, U87cells, U251cells, A172cells, and U373cells and A172cellsMethods We detected expression levels of Zfx mRNA in SHG44cells, U87cells, U251cells, A172cells, and U373cells and A172cells by semi-quantitative RT-PCR. we performed real-time quantitative PCR on35pathologically confirmed glioma samples (Grade Ⅰ-4cases, Grade Ⅱ-13cases, Grade Ⅲ-11cases, and GradeⅣ-7cases) and on5noncancerous brain tissue samples.Result One237bp fragment was amplificated from five human glioblastoma cells by semi-quantitative PCR. We discovered that Zfx mRNA was expressed in U251cells, U87cells, U373cells, and A172cells. The expression level of Zfx is significantly higher in gliomas compared to noncancerous brain tissue.Conclusion The results of our study demonstrate that the Zfx gene is highly expressed in glioma tissue and glioma cell lines. Part Ⅱ:The function of Zfx gene in the the human malignant glioma cell line U251Objective To analyze the influence of Zfx gene in Cell cycle, Cell proliferation, Cell apoptosis of human glioblastoma U251cells.Methods We used lentivirus-mediated small interfering RNAs (siRNAs) to knock down Zfx expression in the human malignant glioma cell line U251. Changes in Zfx target gene expression were determined by real-time RT-PCR. Cell proliferation was examined by a High Content Screening assay. DNA synthesis in proliferating cells was determined by BrdU incorporation. Cell cycle distribution and apoptosis were detected by flowcytometric analysis.Result Using a lentivirus-based RNAi approach, Zfx expression was significantly inhibited in human glioblastoma U251cells. The effects of Zfx knockdown on cell proliferation, cell cycle distribution, and apoptosis were assessed.Conclusion The results of our study demonstrate that the Zfx gene may play a critical role in cell proliferation, cell cycle distribution, and apoptosis of human malignant glioma cells.