The Zfx gene is located on the mammalian X chromosome, at Xp22.12, approximately 23 Mb proximal to this boundary. Zfx is a zinc finger transcription factor that is highly conserved among vertebrates. It contains an acidic transcriptional activation domain, a nuclear localization sequence, and a DNA binding domain consisting of 13 C2H2-type zinc fingers [7]. Zinc finger proteins are characterized by the presence of two cysteines (Cys2) and two histidines (His2) in what

is called a zinc finger domain. This domain stabilizes the three-dimensional structure, consisting of a two-stranded Apoptosis Compound Library cell line antiparallel β-sheet and an α-helix surrounding a central zinc ion [8]. Zinc finger proteins play important roles in multiple biological processes, gene expression, differentiation, and embryonic development [9, 10]. To explore the role of Zfx in human malignant glioma, we began

with an expression analysis of Zfx mRNA in glioma tumors and glioma cell lines. We also used lentivirus-mediated siRNA targeting of Zfx to down-regulate its expression in the human malignant cell line U251 [11]. Finally, we investigated the effect of Zfx silencing on the cell cycle, apoptosis, and proliferation of U251 cells. 2. Materials and methods 2.1 Cell line preparation Human glioma U251 cells, derived from grade IV astrocytomas-glioblastoma multiforme (GBM), and human renal epithelial 293T cells were purchased from Cell Bank Type Culture Collection of Chinese Academy of Sciences (CBTCCCAS, Shanghai, China) and maintained in Dulbecco’s CA3 mw modified Eagle’s medium (DMEM, GIBCO) with 10% fetal bovine serum (FBS, GIBCO) at 37°C in a humidified atmosphere of 5% CO2. 2.2 Clinical sample preparation CX-5461 clinical trial before Ribonucleotide reductase the study began, written informed consent was obtained from all patients who participated in the study, which was approved by the Ethics Committee of SooChow University. All experiments comply with the current

laws of our country. Thirty-five glioma samples were obtained from 35 Chinese patients from March 2009 to Septemper 2010 at the Department of Neurosurgery of The First Affiliated Hospital of Soochow University (Grade I-4cases, Grade II-13cases, Grade III-11cases, and Grade IV-7cases according to the 2007 WHO Classification system). The patients consisted of 19 males and 17 females. The mean ages of the patients at the time of surgery were 38 (male) and 41 (female). All tumors were from patients with newly diagnosed gliomas, who had received no therapy before sample collection. Five adult noncancerous brain tissues were obtained from surgical resections of 5 trauma patients for whom a partial resection of normal brain tissue was required as decompression treatment to reduce increased intracranial pressure under the permission of each patient’s family.