These findings indicate Selleck BV-6 that CENP-H might play an essential role in kinetochore assembly and function throughout the cell cycle. CENP-H is also strongly correlated with human cancer. It’s expression was deregulated in colorectal cancers, and ectopic overexpression of CENP-H induces chromosome instability in diploid cell lines [6]. In addition, CENP-H was deregulated in oral squamous cell carcinomas (SCCs), nasopharyngeal carcinoma (NPC), and esophageal carcinoma [15–17]. The expression of CENP-H might be a valuable prognostic marker which could predict the early stage NPC [15]. Further more, the expression of CENP-H in oral SCCs was significantly correlated

with the cell proliferation in malignant conditions[17]. Genomic aberrations including aneuploidy in epithelial cells of the oral mucosa indicate high risks

of oral BI 10773 cell line cancer and cancer-related mortality [18]. Tongue cancer is one of the most common and serious types of oral cancer with poor prognosis [19, 20]. It is of great clinical value to identify efficient proliferation markers and valuable markers that help to find tongue cancer patients at very early stage. In this study, we investigated the expression of CENP-H in tongue cancer and evaluated the role of CENP-H in proliferation of tongue cancer cells. Inhibitor Library screening Methods Cell cultures Primary cultured normal tongue mucosa epithelial cells (TEC) were maintained in Keratinocyte-SFM (Gibco, Invitrogen Corp, USA). Tongue cancer cell lines TSCCa and Tca8113 were cultured in RPMI 1640 medium supplemented with 10% fetal calf serum (HyClone, Logan, UT). Vectors and retroviral infection Silence endogenous CENP-H, RNAi oligonucleotides (5-GGATCCTGCCCTTAAGGAAAT-3) Calpain was cloned into the pSuper-retro-puro vector to generate pSuper-retro-CENP-H-siRNA. Retroviral production and infection were performed as described previously[21]. Stable Tca8113 cells expressing CENP-H RNAi were selected for 10 days with 0.5 lg/ml

puromycin 48 h after infection. After 10 days selection, the Tca8113 cell lysates prepared from the pooled population of cells in sample buffer were fractionated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the detection of CENP-H protein level. Patients and tissue specimens The present study was performed on 168 cases of paraffin-embedded archived tongue cancer samples obtained from the Department of Pathology, the Second Affiliated Hospital of Sun Yat-sen University (PR China). Prior patients’ consents and approval from the Institutional Research Ethics Committee were obtained for the purpose of research. The final study population included 61 female and 107 male patients (age range, 24–82 years). The median follow-up time for overall survival was 63.14 months (range, 3–169 months) for patients who were still alive at the time of the analysis.