Purpose: To identify epigenetic molecular makers in plasma for the early detection of colorectal cancer. Experimental Design: We retrospectively analyzed the methylation status of 10 genes in fresh-frozen tissues and corresponding plasma samples from 243 patients with stage I and II sporadic colorectal cancer, 276 healthy individuals, and plasma from 64 colorectal adenoma patients using methylation-specific PCR. The methylation score (M score) was used to find molecular markers with high sensitivity and specificity. Results: Of the 243 colorectal cancer tissues, methylation was detected in 18% for p14, 34% for p16, 27% for APC, 34% for DAPK, 32% for HLTF, 21% for hMLH1, 39% for MGMT, 24% for RARβ2, 58% for RASSF2A, and 74% for Wif-1. Receiver operator characteristic curve analysis in plasma from 243 patients with cancer and 276 healthy individuals showed that the M score of any single gene had a sensitivity of <40% after controlling for age, sex, and tumor location. The specificity of the M score was not different between multigene and single gene analyses, but the sensitivity of the M score was significantly increased by multigene analysis. For all patients, the M score in a model including APC, MGMT, RASSF2A, and Wif-1 genes had a sensitivity of 86.5% and a specificity of 92.1% when 1.6 was used as a cutoff. In this model, the M score had a positive predictive value of 90.6% and a negative predictive value of 88.8%. Conclusion: The present study suggests that tumor-specific methylation of APC, MGMT, RASSF2A, and Wif-1 genes might be a valuable biomarker in plasma for the early detection of colorectal cancer. (Clin Cancer Res 2009;15(19):6185-91)

Our study suggests that tumor-specific methylation of the p16, RASSF1A, H-cadherin, and RARbeta genes may be a valuable biomarker for the early detection of NSCLC in bronchial lavage, and that the age-related methylation of FHIT gene in the normal bronchial epithelium is related to the exposure to tobacco smoke.

Objective We documented previously that if study quality is accounted for, evidence from occupational cohort studies on benzene supports a possible association with some lymphoma subtypes, in particular multiple myeloma, and acute and chronic lymphocytic leukemia. Here, we extend these analyses to chronic myeloid leukemia (CML). Methods Three strategies to assess study quality (stratification by the year‐of‐start of follow‐up, stratification by the strength of the reported acute myeloid leukemia (AML) association, and stratification by the quality of benzene exposure assessment) were employed in a meta‐analysis of occupational benzene exposure and CML. We hypothesized that stratification by these study quality dimensions would identify a subgroup of occupational cohort studies that is most informative for the evaluation of the possible association between benzene and CML. Results The overall meta‐relative risk (mRR) was non‐significantly elevated (1.23; 95% confidence interval (CI): 0.93–1.63). The mRRs increased with increasing study quality for all dimensions with a significant elevation for studies with start of follow‐up after 1970 (1.67; 95% CI: 1.02–2.74). The highest study quality stratum for AML significance and exposure quality showed an elevated but non‐significant increased mRR (1.40; 95% CI: 0.86–2.27, and 1.68; 95% CI: 0.74–3.84, respectively). Conclusions Although limited by low statistical power, the current meta‐analysis provides support for a possible association of occupational exposure to benzene and the risk of CML. Am. J. Ind. Med. 55:779–785, 2012. © 2012 Wiley Periodicals, Inc.

Cytoskeleton-associated protein 2 (CKAP2), also known as tumor-associated microtubule-associated protein (TMAP), is a novel microtubule-associated protein that is frequently upregulated in various malignances. However, its cellular functions remain unknown. A previous study has shown that its protein level begins to increase during G 1 /S and peaks at G 2 /M, after which it decreases abruptly. Ectopic overexpression of TMAP/ CKAP2 induced microtubule bundling related to increased microtubule stability. TMAP/CKAP2 overexpression also resulted in cell cycle arrest during mitosis due to a defect in centrosome separation and subsequent formation of a monopolar spindle. We also show that degradation of TMAP/CKAP2 during mitotic exit is mediated by the anaphase-promoting complex bound to Cdh1 and that the KEN box motif near the N terminus is necessary for its destruction. Compared to the wild type, expression of a nondegradable mutant of TMAP/ CKAP2 significantly increased the occurrence of spindle defects and cytokinesis failure. These results suggest that TMAP/CKAP2 plays a role in the assembly and maintenance of mitotic spindles, presumably by regulating microtubule dynamics, and its destruction during mitotic exit serves an important role in the completion of cytokinesis and in the maintenance of spindle bipolarity in the next mitosis.

Fragile histidine triad (FHIT) gene plays an important role in the pathogenesis of lung cancer. However, the clinicopathological significance of CpG island hypermethylation of FHIT gene in non-small cell lung cancer (NSCLC) remains to be elucidated. We studied FHIT methylation in 254 NSCLCs in order to further understand the clinicopathological and prognostic significance of FHIT methylation in NSCLC. Methylation status of the FHIT gene was examined using Methylation-Specific PCR. All statistical analyses were two-sided, with a 5% type I error rate. Hypermethylation of the FHIT gene occurred more frequently in squamous cell carcinoma than adenocarcinoma. For 93 adenocarcinomas there was no statistically significant association between FHIT methylation and age, gender, smoking history, pathologic stage and p16 methylation. However, FHIT methylation in 125 squamous cell carcinomas was associated with exposure to tobacco smoke and p16 methylation, but not with age, gender and pathologic stage. Hypermethylation of FHIT in squamous cell carcinomas occurred more frequently in current smokers (45%) than in never-smokers (13%). FHIT methylation was significantly associated with p16 methylation in current- and ex-smokers (P = 0.02 and P = 0.01, respectively) with squamous cell carcinoma and in patients with pathologic stage I squamous cell carcinoma (P = 0.001). Patients with p16 methylation were 3.74 times [95% confidence interval (CI) = 1.62 - 7.95; P = 0.001] more likely to have FHIT methylation in squamous cell carcinoma. FHIT methylation in squamous cell carcinoma occurred at a 4.62 times (95% CI = 1.26 - 34.97; P = 0.02) higher prevalence in current smokers than in never-smokers. No prognostic effect of FHIT methylation was observed in stage I and stage II NSCLCs. In conclusion, hypermethylation of the FHIT gene did not have a prognostic significance in early stage NSCLCs. The FHIT methylation is associated with the p16 methylation and smoking in squamous cell carcinoma, suggesting that FHIT may cooperate with p16 for the development of squamous cell carcinoma of lung in individuals exposed to tobacco smoke.

Despite advances in the detection and treatment of lung cancer, the prognosis for patients with lung cancer is poor, partly as a result of recurrences. We retrospectively analyzed the relationship between recurrence and survival in patients with nonsmall cell lung cancers (NSCLC), and the promoter methylation of p16, GSTP1, FHIT, H-cadherin, and RARb2 genes to identify a prognostic molecular marker associated with the recurrence of NSCLC. Methylation status from 335 paraffin blocks was determined by methylation-specific PCR. Of the 335 NSCLC samples, promoter methylation was detected in 35% for p16, 39% for RARb2, 42% for H-cadherin, 7% for GSTP1, and 21% for FHIT. Recurrence was observed in 39% (132 of 335) of the patients. Recurrence was significantly associated with histology (P = 0.001) and pathologic stage (P = 0.009). Hypermethylation of any single gene was not associated with recurrence in patients. However, cohypermethylation of p16 and FHIT genes in stage I NSCLCs was associated with an increased risk of recurrence [odds ratio, 6.43; 95% confidence interval (CI), 1.04-20.19; P = 0.02] and poor recurrence-free survival after surgery (hazard ratio, 2.03; 95% CI, 1.09-6.23; P = 0.02). In addition, their survival after recurrence was also 4.62 times poorer (95% CI, 1.27-16.48; P = 0.005) than for those without cohypermethylation of both genes. In conclusion, the present study suggests that cohypermethylation of p16 and FHIT genes in patients with stage I NSCLC may be a valuable biomarker for predicting the recurrence-associated prognosis of the disease. (Cancer Res 2006; 66(8): 4049-54)

When we were studying phosphorylated proteins in the rat brain after electroconvulsive shock (ECS), we observed the rapid phosphorylation of a 75-kDa protein, which cross-reacted with the anti-phospho-p70 S6 kinase antibody. The phosphorylated protein was purified and identified as moesin, a member of the ezrin/radixin/ moesin (ERM) family and a general cross-linker between cortical actin filaments and plasma membranes. The purified moesin from rat brain was phosphorylated at serine and threonine residues. Moesin was rapidly phosphorylated at the threonine 558 residue after ECS in the rat hippocampus, peaked at 1 min, and returned to the basal level by 2 min after ECS. To investigate the mechanism of moesin phosphorylation in neuronal cells, we stimulated a rat hippocampal progenitor cell, H19 -7/ IGF-IR, with glutamate, and observed the increased phosphorylation of moesin at Thr-558. Glutamate transiently activated RhoA, and constitutively active RhoA increased the basal level phosphorylation of moesin. The inhibition of RhoA and its effector, Rho kinase, abolished increased Thr-558 phosphorylation by glutamate in H19 -7/IGF-IR cells, suggesting that the phosphorylation of moesin at Thr-558 in H19 -7/IGF-IR cells by glutamate is mediated by RhoA and Rho kinase activation. Electroconvulsive shock (ECS)1 has been used for the treatments of psychiatric disorders such as depression, schizophrenia, and manic-depressive illness. Although the mechanism of the treatment has not been elucidated, recent progress in the cellular and molecular biology of neuronal tissues has enabled neuronal functions to be linked to the regulation of gene expression and intracellular signal transduction (1). Previously, we reported (4) that ECS induces the expressions of various immediate early genes such as c-fos, junB, and TiS-8 and that it suppresses the expression of inositol 1,4,5-triphosphate kinase and inositol 1,4,5-triphosphate receptor genes. We also observed (2-5, 7) that ECS caused the phosphorylation of many signaling molecules, including those in the Pyk2-Ras-Raf-MEK-ERK pathway, and of stress-signaling pathways in various rat brain regions.Earlier we reported that ECS induces the phosphorylation of CREB in the rat hippocampus (6). Several protein kinases have been suggested to be CREB kinases in neuronal tissues, and we looked for protein kinases potentially responsible for CREB phosphorylation. When we were studying the phosphorylation of one of the suggested CREB kinases, ribosomal S6 kinase (S6K), we observed that a 75-kDa protein in the rat hippocampus was rapidly phosphorylated after ECS. This was detected by an antibody specific to phospho-p70 S6K. The protein was not detected by the anti-p70 S6K antibody, suggesting that this protein was detected nonspecifically by an anti-phospho-p70-S6K antibody. So we purified and identified this protein as moesin (membrane-organizing extension spike protein), a member of the ezrin/radixin/moesin (ERM) family of proteins.

When these points were taken together, we concluded that CKAP2 is up-regulated in primary human gastric adenocarcinomas at high frequency and might be useful for diagnosing and discriminating adenocarcinomas from tubular adenomas of the stomach.