Porphyromonas gingivalis Impairs Oral Epithelial Barrier through Targeting GRHL2.

Oral mucosa provides the first line of defense against a diverse array of environmental and microbial irritants by forming the barrier of epithelial cells interconnected by multiprotein tight junctions (TJ), adherens junctions, desmosomes, and gap junction complexes. Grainyhead-like 2 (GRHL2), an epithelial-specific transcription factor, may play a role in the formation of the mucosal epithelial barrier, as it regulates the expression of the junction proteins. The current study investigated the role of GRHL2 in the Porphyromonas gingivalis (Pg)-induced impairment of epithelial barrier functions. Exposure of human oral keratinocytes (HOK-16B and OKF6 cells) to Pg or Pg-derived lipopolysaccharides (Pg LPSs) led to rapid loss of endogenous GRHL2 and the junction proteins (e.g., zonula occludens, E-cadherin, claudins, and occludin). GRHL2 directly regulated the expression levels of the junction proteins and the epithelial permeability for small molecules (e.g., dextrans and Pg bacteria). To explore the functional role of GRHL2 in oral mucosal barrier, we used a Grhl2 conditional knockout (KO) mouse model, which allows for epithelial tissue-specific Grhl2 KO in an inducible manner. Grhl2 KO impaired the expression of the junction proteins at the junctional epithelium and increased the alveolar bone loss in the ligature-induced periodontitis model. Fluorescence in situ hybridization revealed increased epithelial penetration of oral bacteria in Grhl2 KO mice compared with the wild-type mice. Also, blood loadings of oral bacteria (e.g., Bacteroides, Bacillus, Firmicutes, ß-proteobacteria, and Spirochetes) were significantly elevated in Grhl2 KO mice compared to the wild-type littermates. These data indicate that Pg bacteria may enhance paracellular penetration through oral mucosa in part by targeting the expression of GRHL2 in the oral epithelial cells, which then impairs the epithelial barrier by inhibition of junction protein expression, resulting in increased alveolar tissue destruction and systemic bacteremia.

Human Papillomavirus 16 E6 Induces FoxM1B in Oral Keratinocytes through GRHL2.

High-risk human papillomavirus (HPV) is a major risk factor for oral and pharyngeal cancers (OPCs), yet the detailed mechanisms by which HPV promotes OPCs are not understood. Forkhead box M1B (FoxM1B) is an oncogene essential for cell cycle progression and tumorigenesis, and it is aberrantly overexpressed in many tumors. We previously showed that FoxM1B was the putative target of an epithelial-specific transcription factor, Grainyhead-like 2 (GRHL2). In the current study, we demonstrate that HPV type 16 (HPV-16) E6 induces FoxM1B in human oral keratinocytes (HOKs) and tonsillar epithelial cells (TECs) in part through GRHL2. FoxM1B was barely detectable in cultured normal human oral keratinocytes (NHOKs) and progressively increased in immortalized HOKs harboring HPV-16 genome (HOK-16B) and tumorigenic HOK-16B/BaP-T cells. Retroviral expression of HPV-16 E6 and/or E7 in NHOKs, TECs, and hypopharyngeal carcinoma cells (FaDu) revealed induction of FoxM1B and GRHL2 by the E6 protein but not E7. Both GRHL2 and FoxM1B were strongly induced in the epidermis of HPV-16 E6 transgenic mice and HPV+ oral squamous cell carcinomas. Ectopic expression of FoxM1B led to acquisition of transformed phenotype in HOK-16B cells. Loss of FoxM1B by lentiviral short hairpin RNA vector or chemical inhibitor led to elimination of tumorigenic characteristics of HOK-16B/BaP-T cells. Luciferase reporter assay revealed that GRHL2 directly bound and regulated the FoxM1B gene promoter activity. Using epithelial-specific Grhl2 conditional knockout mice, we exposed wild-type (WT) and Grhl2 KO mice to 4-nitroquinolin 1-oxide (4-NQO), which led to induction of FoxM1B in the tongue tissues and rampant oral tumor development in the WT mice. However, 4-NQO exposure failed to induce tongue tumors or induction of FoxM1B expression in Grhl2 KO mice. Collectively, these results indicate that HPV-16 induces FoxM1B in part through GRHL2 transcriptional activity and that elevated FoxM1B level is required for oropharyngeal cancer development.

Effects of Bioactive Compounds on Odontogenic Differentiation and Mineralization.

Direct pulp capping involves the placement of dental materials directly onto vital pulp tissues after deep caries removal to stimulate the regeneration of reparative dentin. This physical barrier will serve as a "biological seal" between these materials and the pulp tissue. Although numerous direct pulp capping materials are available, the use of small bioactive compounds that can potently stimulate and expedite reparative dentin formation is still underexplored. Here, the authors compared and evaluated the pro-osteogenic and pro-odontogenic effects of 4 small bioactive compounds- phenamil (Phen), purmorphamine (Pur), genistein (Gen), and metformin (Met). The authors found that these compounds at noncytotoxic concentrations induced differentiation and mineralization of preosteoblastic MC3T3-E1 cells and preodontoblastic dental pulp stem cells (DPSCs) in a dose-dependent manner. Among them, Phen consistently and potently induced differentiation and mineralization in vitro. A single treatment with Phen was sufficient to enhance the mineralization potential of DPSCs in vitro. More importantly, Phen-treated DPSCs showed enhanced odontogenic differentiation and mineralization in vivo. Our study suggests that these small bioactive compounds merit further study for their potential clinical use as pulp capping materials.

Epigenetic gene regulation by histone demethylases: emerging role in oncogenesis and inflammation.

Histone N-terminal tails of nucleosomes are the sites of complex regulation of gene expression through post-translational modifications. Among these modifications, histone methylation had long been associated with permanent gene inactivation until the discovery of Lys-specific demethylase (LSD1), which is responsible for dynamic gene regulation. There are more than 30 members of the Lys demethylase (KDM) family, and with exception of LSD1 and LSD2, all other KDMs possess the Jumonji C (JmjC) domain exhibiting demethylase activity and require unique cofactors, for example, Fe(II) and α-ketoglutarate. These cofactors have been targeted when devising KDM inhibitors, which may yield therapeutic benefit. KDMs and their counterpart Lys methyltransferases (KMTs) regulate multiple biological processes, including oncogenesis and inflammation. KDMs' functional interactions with retinoblastoma (Rb) and E2 factor (E2F) target promoters illustrate their regulatory role in cell cycle progression and oncogenesis. Recent findings also demonstrate the control of inflammation and immune functions by KDMs, such as KDM6B that regulates the pro-inflammatory gene expression and CD4+ T helper (Th) cell lineage determination. This review will highlight the mechanisms by which KDMs and KMTs regulate the target gene expression and how epigenetic mechanisms may be applied to our understanding of oral inflammation.

The Role of ORAI1 in the Odontogenic Differentiation of Human Dental Pulp Stem Cells.

Pulp capping, or placing dental materials directly onto the vital pulp tissues of affected teeth, is a dental procedure that aims to regenerate reparative dentin. Several pulp capping materials are clinically being used, and calcium ion (Ca(2+)) released from these materials is known to mediate reparative dentin formation. ORAI1 is an essential pore subunit of store-operated Ca(2+) entry (SOCE), which is a major Ca(2+) influx pathway in most nonexcitable cells. Here, we evaluated the role of ORAI1 in mediating the odontogenic differentiation and mineralization of dental pulp stem cells (DPSCs). During the odontogenic differentiation of DPSCs, the expression of ORAI1 increased in a time-dependent manner. DPSCs knocked down with ORAI1 shRNA (DPSC/ORAI1sh) or overexpressed with dominant negative mutant ORAI1(E106Q) (DPSC/E106Q) exhibited the inhibition of Ca(2+) influx and suppression of odontogenic differentiation and mineralization as demonstrated by alkaline phosphatase (ALP) activity/staining as well as alizarin red S staining when compared with DPSCs of their respective control groups (DPSC/CTLsh and DPSC/CTL). The gene expression for odontogenic differentiation markers such as osteocalcin, bone sialoprotein, and dentin matrix protein 1 (DMP1) was also suppressed. When DPSC/CTL or DPSC/E106Q cells were subcutaneously transplanted into nude mice, DPSC/CTL cells induced mineralized tissue formation with significant increases in ALP and DMP1 staining in vivo, whereas DPSC/E106Q cells did not. Collectively, our data showed that ORAI1 plays critical roles in the odontogenic differentiation and mineralization of DPSCs by regulating Ca(2+) influx and that ORAI1 may be a therapeutic target to enhance reparative dentin formation.

Pulp-dentin Regeneration: Current State and Future Prospects.

The goal of regenerative endodontics is to reinstate normal pulp function in necrotic and infected teeth that would result in reestablishment of protective functions, including innate pulp immunity, pulp repair through mineralization, and pulp sensibility. In the unique microenvironment of the dental pulp, the triad of tissue engineering would require infection control, biomaterials, and stem cells. Although revascularization is successful in resolving apical periodontitis, multiple studies suggest that it alone does not support pulp-dentin regeneration. More recently, cell-based approaches in endodontic regeneration based on pulpal mesenchymal stem cells (MSCs) have demonstrated promising results in terms of pulp-dentin regeneration in vivo through autologous transplantation. Although pulpal regeneration requires the cell-based approach, several challenges in clinical translation must be overcome-including aging-associated phenotypic changes in pulpal MSCs, availability of tissue sources, and safety and regulation involved with expansion of MSCs in laboratories. Allotransplantation of MSCs may alleviate some of these obstacles, although the long-term stability of MSCs and efficacy in pulp-dentin regeneration demand further investigation. For an alternative source of MSCs, our laboratory developed induced MSCs (iMSCs) from primary human keratinocytes through epithelial-mesenchymal transition by modulating the epithelial plasticity genes. Initially, we showed that overexpression of ΔNp63α, a major isoform of the p63 gene, led to epithelial-mesenchymal transition and acquisition of stem characteristics. More recently, iMSCs were generated by transient knockdown of all p63 isoforms through siRNA, further simplifying the protocol and resolving the potential safety issues of viral vectors. These cells may be useful for patients who lack tissue sources for endogenous MSCs. Further research will elucidate the level of potency of these iMSCs and assess their transdifferentiation capacities into functional odontoblasts when transplanted into the root canal microenvironment.

Impact of guideline adherence on patient outcomes in early-stage epithelial ovarian cancer.

AIM: The aim of this study was to evaluate the effects of adherence to National Comprehensive Cancer Network (NCCN) guidelines on survival outcomes in patients with early-stage epithelial ovarian cancer. METHODS: Our institutional cancer registry data on 266 patients with Stage I epithelial ovarian cancer was reviewed retrospectively and compliance with treatment guidelines for surgery and adjuvant treatment was determined. Patients were categorized according to adherence or non-adherence. The primary endpoints were recurrence-free survival and disease-specific survival. Hazard ratios (HRs) for survival were estimated with a Cox proportional hazards model. RESULTS: Of the 266 patients, 71 (26.7%) underwent adequate surgical staging in accordance with the guidelines. The guidelines for adjuvant chemotherapy were followed adequately in all 71 patients that were adherent to surgical staging and in 163 of the 195 patients with non-adherence to surgical staging (83.6%). Multivariate analysis, adjusted for prognostic factors, identified higher recurrence-free survival (HR, 0.36; 95% CI, 0.15-0.88) and disease-specific survival (HR, 0.42; 95% CI, 0.16-1.12) among patients whose treatment adhered to both surgical and chemotherapy guidelines, although disease-specific survival was not statistically significant. When excluding clear cell histology from the cohort, the guideline-adherent group had significantly better disease-specific survival than the non-adherent group (HR, 0.13; 95% CI, 0.02-0.94). CONCLUSION: The results of this study suggest that adherence to NCCN guidelines may improve survival outcomes in patients with early-stage epithelial ovarian cancer, particularly in cases other than clear cell histology.

Usefulness of meniscal width to transverse diameter ratio on coronal MRI in the diagnosis of incomplete discoid lateral meniscus.

AIM: To evaluate the clinical utility of the meniscal width to transverse diameter ratio (L/M ratio) of the lateral meniscus in the diagnosis of incomplete discoid lateral meniscus (IDLM) as compared with the arthroscopic diagnosis, meniscal width to tibial diameter ratio (L/T ratio) and conventional lateral meniscus width criteria. MATERIALS AND METHODS: This retrospective study sample included 41 patients with IDLM who underwent knee magnetic resonance imaging (MRI) and arthroscopy, as well as 50 controls with normal lateral menisci. MRI examinations were interpreted independently by two radiologists, both of whom were blinded to clinical information and radiological reports. Assessment of meniscal width (L), maximal transverse diameter of the lateral meniscus (M), and transverse diameter of the tibia (T) was carried out on central coronal sections that were observed to pass through the medial collateral ligament. L/M and L/T ratios were calculated. These results were correlated with arthroscopic findings and analysed statistically using categorical regression analysis and non-parametric correlation analysis. Using arthroscopic findings as the standard of reference, sensitivity and specificity were calculated for: (1) 12, 13, 14, and 15 mm meniscal width thresholds; (2) 40%, 50%, 60%, and 70% L/M ratio thresholds; and (3) 15%, 18%, 20%, and 25% L/T ratio thresholds. RESULTS: The mean L/M ratio of the IDLM was approximately 67% and was statistically significantly higher than the control (44%). The best diagnostic discrimination was achieved using a threshold of 50%. The mean L/T ratio of the IDLM was approximately 23% and was statistically significant. The best diagnostic discrimination was achieved using a threshold of 18%. The threshold of 13 mm of meniscal width also showed high sensitivity and high specificity. CONCLUSION: The use of the L/M ratio or L/T ratio in combination with meniscal width criteria may be a useful method for evaluating IDLM.

Impact of intraoperative rupture of the ovarian capsule on prognosis in patients with early-stage epithelial ovarian cancer: a meta-analysis.

BACKGROUND: The impact of intraoperative rupture on prognosis is controversial in early-stage epithelial ovarian cancer (EOC). Thus, we performed a meta-analysis to determine its impact and to evaluate factors to increase its risk. METHODS: We searched PubMed, Embase, and the Cochrane Library till May 2011, and 9 eligible studies including 2382 patients were evaluated. All patients were classified into three groups: no rupture; intraoperative rupture; preoperative involvement. RESULTS: Preoperative involvement decreased progression-free survival when compared with intraoperative rupture (PFS; HR, 1.47; 95% CI, 1.01-2.14), which also showed poorer PFS than no rupture (HR, 2.41; 95% CI, 1.74-3.33). Although preoperative involvement reduced PFS when compared with intraoperative rupture (HR, 2.63; 95% CI, 1.11-6.20), there was no difference in it between intraoperative rupture and no rupture in patients who underwent complete surgical staging operation and adjuvant platinum-based chemotherapy if needed (HR, 1.49; 95% CI, 0.45-4.95). Furthermore, adhesion to adjacent tissues, grade 2 or 3 disease were more common (ORs, 2.01 and 2.47; 95% CIs, 1.20-3.37 and 1.12-5.46), whereas mucinous tumor was less frequent (OR, 0.51; 95% CI, 0.37-0.72) in intraoperative rupture than in no rupture. CONCLUSIONS: Intraoperative rupture may not decrease PFS when compared with no rupture in patients with early-stage EOC who underwent complete surgical staging operation and adjuvant platinum-based chemotherapy. Furthermore, more adhesion to adjacent tissues and grade 2 or 3 disease, and less mucinous tumor are expected to increase the risk of intraoperative rupture.

Efficacy of neoadjuvant chemotherapy in patients with FIGO stage IB1 to IIA cervical cancer: an international collaborative meta-analysis.

BACKGROUND: The efficacy of neoadjuvant chemotherapy before surgery (NCS) has not been well-established in FIGO stage IB1 to IIA cervical cancer when compared with primary surgical treatment (PST). Thus, we performed a meta-analysis to determine the efficacy of NCS in patients with FIGO stage IB1 to IIA cervical cancer when compared with PST. METHODS: We searched Pubmed, Embase and the Cochrane Library between January 1987 and September 2010. Since there was a relative lack of relevant randomized controlled trials (RCTs), we included 5 RCTs and 4 observational studies involving 1784 patients among 523 potentially relevant studies. RESULTS: NCS was related with lower rates of large tumor size (≥4 cm) (ORs, 0.22 and 0.10; 95% CI, 0.13-0.39 and 0.02-0.37) and lymph node metastasis (ORs, 0.61 and 0.38; 95% CI, 0.37-0.99 and 0.20-0.73) than PST in all studies and RCTs. Furthermore, NCS reduced the need of adjuvant radiotherapy (RT) in all studies (OR, 0.57; 95% CI, 0.33-0.98), and distant metastasis in all studies and RCTs (ORs, 0.61 and 0.61; 95% CI, 0.42-0.89 and 0.38-0.97). However, overall and loco-regional recurrences and progression-free survival were not different between the 2 treatments. On the other hand, NCS was associated with poorer overall survival in observational studies when compared with PST (HR, 1.68; 95% CI, 1.12-2.53). CONCLUSIONS: Although NCS reduced the need of adjuvant RT by decreasing tumor size and lymph node metastasis, and distant metastasis, it failed to improve survival when compared with PST in patients with FIGO stage IB1 to IIA cervical cancer.

Metabolic syndrome is associated with increased risk of recurrent colorectal adenomas in Korean men.

OBJECTIVE: Because of the high incidence of recurrent colorectal adenomas, regular surveillance by colonoscopy is recommended. However, there is still a shortage of information on the factors that influence the incidence of recurrent colorectal adenomas in patients with a history of these lesions. The aim of this study was to determine the association between the development of recurrent colorectal adenomas, metabolic syndrome and obesity. SUBJECTS AND METHODS: The hospital-based cohort was composed of 193 patients who had recurrent colorectal adenomas removed between January 2002 and December 2003. The Cox proportional hazard model was used to determine hazard ratio (HR) and 95% confidence interval (CI) between obesity, metabolic syndrome and other factors, and the incidence of recurrent adenomatous polyps. RESULTS: The mean follow-up period was 4.8 person-years. In all, 78 of the patients (40.4%) had recurrent colorectal adenomas. In the overall recurrent adenoma group, significant associations between metabolic syndrome (HR, 1.33; 95% CI, 1.02-1.73), waist circumference (WC) ≥ 90 cm (HR, 1.42; 95% CI, 1.06-1.90) and waist-hip ratio (WHR) ≥ 0.9 (HR, 2.03; 95% CI, 1.55-2.68) were found. Moreover, advanced adenomas were significantly associated with metabolic syndrome (HR, 2.81; 95% CI, 1.86-4.25), body mass index ≥ 25 kg m(-2) (HR, 2.69; 95% CI, 1.64-4.42), WC (HR, 2.16; 95% CI, 1.31-3.54) and WHR (HR, 1.99; 95% CI, 1.28-3.11). In addition, current smoking (HR, 2.60; 95% CI, 1.09-6.25) and alcohol consumption (HR, 2.20; 95% CI, 1.10-4.39) were also significantly associated with recurrent advanced adenoma. CONCLUSION: Metabolic syndrome and obesity were significantly associated with the development of recurrent colorectal adenomas in Korean adult males. Furthermore, these associations were more strongly associated with advanced adenomas.

Accuracy of MRI findings in chronic lateral ankle ligament injury: comparison with surgical findings.

AIM: To evaluate the accuracy of magnetic resonance imaging (MRI) findings in chronic lateral ankle ligament injury in comparison with that of surgical findings. MATERIALS AND METHODS: Forty-eight cases (25 men, 23 women, mean age 36 years) of clinically suspected chronic ankle ligament injury underwent MRI studies and surgery. Sagittal, coronal, and axial, T1-weighted, spin-echo, proton density and T2-weighted, fast spin-echo images with fat saturation were obtained in all patients. MRI examinations were read in consensus by two fellowship-trained academic musculoskeletal radiologists who evaluated the lateral ankle ligaments, including the anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) without clinical information. The results of the MRI studies were then compared with the surgical findings. RESULTS: The MRI findings of ATFL injury showed a sensitivity of detection of complete tears of 75% and specificity of 86%. The sensitivity of detection of partial tears was 75% and the specificity was 78%. The sensitivity of detection of sprains was 44% and the specificity was 88%. Regarding the MRI findings of CFL injury, the sensitivity of detection of complete tears was 50% and the specificity was 98%. The sensitivity of detection of partial tear was 83% and the specificity was 93%. The sensitivity of detection of sprains was 100% and the specificity was 90%. Regarding the ATFL, the accuracies of detection were 88, 58, 77, and 85% for no injury, sprain, partial tear, and complete tear, respectively, and for the CFL the accuracies of detection were 90, 90, 92, and 96% for no injury, sprain, partial tear, and complete tear, respectively. CONCLUSIONS: The diagnosis of a complete tear of the ATFL on MRI is more sensitive than the diagnosis of a complete tear of the CFL. MRI findings of CFL injury are diagnostically specific but are not sensitive. However, only normal findings and complete tears were statistically significant between ATFL and CFL (p < 0.001).

Bisphosphonates induce senescence in normal human oral keratinocytes.

Bisphosphonate-related osteonecrosis of the jaw (BRONJ) commonly occurs in individuals receiving bisphosphonates (BPs) with clinical manifestations of the exposed necrotic bone. Although defective wound healing of soft tissue is frequently, if not always, observed in BRONJ, the effects of BPs on oral soft tissue or cells remain unknown. To investigate the effects of BPs on cells of oral mucosal tissue, we studied the effect of pamidronate (PAM), one of the BPs most commonly administered to cancer patients, on the phenotypes of normal human oral keratinocytes (NHOK) and fibroblasts (NHOF). When exposed to PAM at 10 µM, NHOK, not NHOF, underwent senescence: NHOK overexpressed senescence-associated ß-galactosidase (SA-ß-Gal), p16INK4A, IL-6, and IL-8. When exposed to a higher level (50 µM) of PAM, NHOK maintained senescent phenotypes, but NHOF underwent apoptosis. PAM-induced senescence in NHOK is mediated, in part, via geranylgeranylation of the mevalonate pathway. Our in vitro 3D oral mucosal tissue construction studies further demonstrated that PAM induced senescence and impaired re-epithelialization of oral mucosa. Analysis of these data indicates that premature senescence of oral mucosal cells and subsequent defective soft-tissue wound healing might be partly responsible for the development of BRONJ in individuals receiving PAM or other BPs.

Neutrophil to lymphocyte ratio for preoperative diagnosis of uterine sarcomas: a case-matched comparison.

BACKGROUND: Uterine sarcomas are rare among all uterine malignancies, and frequently misdiagnosed as benign uterine diseases such as leiomyoma and adenomyosis because of lack of feasible tools for the preoperative diagnosis. Although some studies have suggested the role of serum CA-125 levels for the preoperative diagnosis, the efficacy is controversial. Since malignancy is known to be associated with systemic inflammation which leads to hematological alteration, we compared the efficacy for the preoperative diagnosis of uterine sarcomas between the neutrophil to lymphocyte ratio (NLR) and serum CA-125 levels using a case-match comparison. METHODS: From November 2004 to December 2008, 55 patients with carcinosarcoma (n=21), leiomyosarcoma (n=20) and endometrial stromal sarcoma (n=14) were matched to 330 patients with leiomyoma (n=165) and adenomyosis (n=165) in terms of age at diagnosis, body mass index and uterine volume. RESULTS: The receiver operating characteristic curve showed the best cut-off values of the NLR (>or=2.12) and serum CA-125 levels (>or=27.5U/ml) for the preoperative diagnosis of uterine sarcomas, demonstrating that the NLR was more powerful for the preoperative diagnosis of uterine sarcomas than serum CA-125 levels (sensitivity, 74.5% vs. 52.3%; specificity, 70.3% vs. 50.5%; positive predictive value, 29.5% vs. 15.1%; negative predictive value, 94.3% vs. 86.5%; accuracy, 60.6% vs. 49.6%; p<0.05). Furthermore, the NLR reflected recurrence and progression more accurately than serum CA-125 levels in patients with uterine sarcomas. CONCLUSIONS: These findings suggest that the NLR may be more useful than serum CA-125 levels as a cost-effective tool for the preoperative diagnosis in patients with uterine sarcomas.

Eosinophilic cystitis associated with eosinophilic enterocolitis: case reports and review of the literature.

We report three cases of eosinophilic cystitis. Contrast-enhanced computed tomography (CT) revealed characteristic bladder wall thickening exceeding 10 mm, with preservation of the mucosal lining and intense, progressive contrast enhancement on sequential arterial and delayed scans. Eosinophilic cystitis might have been associated with eosinophilic infiltration in other organs, such as the gastrointestinal tracts and liver.

BubR1 as a prognostic marker for recurrence-free survival rates in epithelial ovarian cancers.

BACKGROUND: Epithelial ovarian cancer is one of the most lethal malignancies, and has a high recurrence rate. Thus, prognostic markers for recurrence are crucial for the care of ovarian cancer. As ovarian cancers frequently exhibit chromosome instability, we aimed at assessing the prognostic significance of two key mitotic kinases, BubR1 and Aurora A. METHODS: We analysed paraffin-embedded tissue sections from 160 ovarian cancer patients whose clinical outcomes had been tracked after first-line treatment. RESULTS: The median recurrence-free survival in patients with a positive and negative expression of BubR1 was 27 and 83 months, respectively (P<0.001). A positive BubR1 expression was also associated with advanced stage, serous histology and high grade. In contrast, Aurora A immunostaining did not correlate with any of the clinical parameters analysed. CONCLUSION: BubR1, but not Aurora A, is a prognostic marker for recurrence-free survival rates in epithelial ovarian cancers.

Clinical impact of under-diagnosis by frozen section examination is minimal in borderline ovarian tumors.

AIMS: The purpose of this study was to examine the clinical impact of under-diagnosis by frozen section examination in borderline ovarian tumors (BOTs). METHODS: We reviewed 209 consecutive patients with BOTs who were diagnosed and treated at our institute between March 1988 and July 2007. Tumors from 182 of 209 patients were evaluated by frozen section examination. After excluding a case with a deferred diagnosis, the results of frozen section examinations were compared with the final paraffin section examination. In 61 patients, the frozen section examination under-diagnosed a BOT as a benign tumor. In 120 patients, the frozen section examination correctly diagnosed or over-diagnosed a BOT. The clinical impact of under-diagnosis was evaluated by comparing the extent of surgery and treatment outcome between the patients who were under-diagnosed (study group) and the patients who were not under-diagnosed (control group). RESULTS: The study group had more mucinous (P=0.03) and/or stage 1A (P=0.02) tumors than the control group. Fewer patients in the study group received adjuvant chemotherapy than the patients in the control group (P=0.02). Fewer patients in the study group underwent radical surgery than the patients in the control group (P=0.02). However, the rates of treatment failure were similar between the two groups (no treatment failure in the study group and seven treatment failures in the control group; P=0.10). CONCLUSIONS: The under-diagnosis by frozen section examination did not compromise the outcome in patients with BOTs, although under-diagnosis was associated with more conservative surgery.

The role of serum CA-125 levels in early-stage epithelial ovarian cancer on preoperative CT and MRI.

BACKGROUND: We sought to identify the role of serum CA-125 levels in early-stage epithelial ovarian cancer (EOC) on preoperative CT and MRI. METHODS: Clinical data of 101 patients with early-stage EOC on preoperative CT and MRI were collected between January 2000 and December 2007. Clinical stage I (n=59) was defined as tumor limited to the ovaries with or without ascites, whereas clinical stage II (n=42) was defined as tumor within the pelvis with or without ascites. The primary endpoint was to investigate the efficacy of serum CA-125 levels for the prediction of advanced-stage disease, and secondary endpoints were to evaluate the accuracy of preoperative CT and MRI, and to examine the role of serum CA-125 levels as a prognostic factor for survival. RESULTS: The results of preoperative CT and MRI were concordant with no peritoneal implants outside the pelvis in 50/101 (50%) and no lymph node metastasis in 71/101 (70%) patients. The receiver operating characteristic curves showed that best cut-off values of serum CA-125 levels were 320 U/ml (71% sensitivity, 84% specificity) and 510 U/ml (67% sensitivity, 80% specificity) for the prediction of peritoneal implants outside the pelvis and lymph node metastasis. The serum CA-125 level (> or =320 U/ml) was a significant factor for the prediction of advanced-stage disease (adjusted OR, 7.43; 95% CI, 2.39-23.04). However, it was not an independent prognostic factor for survival. CONCLUSIONS: Serum CA-125 levels may be very useful for the prediction of advanced-stage disease in early-stage EOC on preoperative CT and MRI.

Regulation of the hTERT promoter activity by MSH2, the hnRNPs K and D, and GRHL2 in human oral squamous cell carcinoma cells.

Higher expression of human telomerase reverse transcriptase (hTERT) and subsequent activation of telomerase occur during cellular immortalization and are maintained in cancer cells. To understand the mode of hTERT expression in cancer cells, we identified cancer-specific trans-regulatory proteins that interact with the hTERT promoter, using the promoter magnetic precipitation assay coupled with mass spectrometry. The identified proteins include MutS homolog 2 (MSH2), heterogeneous nuclear ribonucleoprotein (hnRNP) D, hnRNP K and grainyhead-like 2 (GRHL2). We noticed a higher expression of these proteins in human oral squamous cell carcinoma (OSCC) cells than in normal cells, which do not exhibit telomerase activity. Knockdown of MSH2, hnRNP D and GRHL2 resulted in a notable reduction of the hTERT promoter activity in tested cancer cells. Silencing of the above genes resulted in a significant reduction of the telomerase activity in OSCC cells. Interestingly, among the four identified genes, silencing of GRHL2 was essential in reducing telomerase activity and viability of tested cancer cells. These results suggest a possible role of GRHL2 in telomerase activation during cellular immortalization.