title: The Zfx gene is expressed in human gliomas and is important in the proliferation and apoptosis of the human malignant glioma cell line U251.
abstract: BACKGROUND
  Zfx is a zinc finger protein of the Zfy family, whose members are highly conserved in vertebrates. Zfx is a shared transcriptional regulator of both embryonic stem cells (ESC) and hematopoietic stem cells (HSC), which suggests a common genetic basis of self-renewal in embryonic and adult stem cells. The level of Zfx expression correlates with aggressiveness and severity in many cancer types, including prostate cancer, breast cancer, and leukemia. However, the importance of Zfx in human glioma is largely unknown. In the present study, we examined the role of Zfx in human glioma.
  
  
  METHODS
  We detected expression levels of Zfx mRNA in U251 cells, U87 cells, U373 cells, and A172 cells by semi-quantitative RT-PCR. To analyze the expression of Zfx mRNA in glioma tissues, we performed real-time quantitative PCR on 35 pathologically confirmed glioma samples (Grade I-4cases, Grade II-13cases, Grade III-11cases, and Grade IV-7cases) and on 5 noncancerous brain tissue samples. 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.
  
  
  RESULTS
  We discovered that Zfx mRNA was expressed in U251 cells, U87 cells, U373 cells, and A172 cells. The expression level of Zfx is significantly higher in gliomas compared to noncancerous brain tissue. Using a lentivirus-based RNAi approach, Zfx expression was significantly inhibited in human glioblastoma U251 cells. The effects of Zfx knockdown on cell proliferation, cell cycle distribution, and apoptosis were assessed. Inhibition of Zfx expression in U251 cells by RNAi significantly impaired cell proliferation, increased apoptosis, and arrested cells in S phase.
  
  
  CONCLUSIONS
  The results of our study demonstrate that the Zfx gene is highly expressed in glioma tissue and in glioma cell lines. Furthermore, Zfx may play a critical role in cell proliferation, cell cycle distribution, and apoptosis of human malignant glioma cells.
authors.affiliation: Neurosurgery & Brain and Nerve Research Laboratory, The First Affiliated Hospital of Soochow University, suzhou 215006, China.
authors.forename: Youxin
authors.initials: Y
authors.lastname: Zhou
authors.forename: Zuopeng
authors.initials: Z
authors.lastname: Su
authors.forename: Yulun
authors.initials: Y
authors.lastname: Huang
authors.forename: Ting
authors.initials: T
authors.lastname: Sun
authors.forename: Sansong
authors.initials: S
authors.lastname: Chen
authors.forename: Tingfeng
authors.initials: T
authors.lastname: Wu
authors.forename: Guilin
authors.initials: G
authors.lastname: Chen
authors.forename: Xueshun
authors.initials: X
authors.lastname: Xie
authors.forename: Bin
authors.initials: B
authors.lastname: Li
authors.forename: Ziwei
authors.initials: Z
authors.lastname: Du
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D048708:Cell Growth Processes / Q000235:genetics; D045744:Cell Line, Tumor; D005910:Glioma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D051741:Kruppel-Like Transcription Factors / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics*
chemical_list: D051741:Kruppel-Like Transcription Factors; D012333:RNA, Messenger; C062076:zinc finger protein, X-linked