Diagnostic value of the dual-luciferase report assay for predicting response to glucocorticoid in children with acute lymphoblastic leukemia.

OBJECTIVE: Resistance to glucocorticoid (GC) is a significant clinical problem in some cases of acute lymphoblastic leukemia (ALL). Current methods of assessing GC resistance are time consuming and have limited reproducibility; in this study, we sought to define a new method of evaluating GC sensitivity and resistance in vitro. METHODS: Based on the mechanisms of GC resistance, we hypothesized that the dual-luciferase report (DLR) assay could reflect the transcription effects of GC downstream of the GC-glucocorticoid receptor signaling pathway, thereby allowing the evaluation of reactions to GC. Sixty-two patients with differential GC response were included in this study. The prednisone induction test was used to divide the children with ALL into two groups: GC sensitive (GCS) and GC resistant (GCR). DLR assay was later conducted on those patients to evaluate its value for diagnosis of the GC reactivity. Receiver operating characteristic curves were used to identify the optimal assay cutoff for identifying response to GC. RESULTS: Using the DLR assay analysis, we found that GCR subjects showed significantly lower reporter/control ratios for luciferase, as compared with GCS subjects. The optimal cutoff value for GC response was 0.67, with sensitivity of 77.1% and specificity of 93.3%. The DLR assay results were consistent with prednisone induction test results. Further, the DLR assay was simpler, more sensitive, and less time-consuming than the prednisone induction test. CONCLUSIONS: Our study showed that the DLR assay is relatively fast, simple, and sensitive. Accordingly, it could be useful for detecting GC response in children with ALL.

Cumulative methylation alternations of gene promoters and protein markers for diagnosis of epithelial ovarian cancer.

DNA methylation plays an important role in carcinogenesis and cancer development. In this study, we examined gene methylation in epithelial ovarian cancer (EOC) using cationic conjugated polymer (CCP)-based fluorescence resonance energy transfer (FRET) to evaluate the application of cumulative methylation alternations of genes associated with cancer antigen 125 for early cancer diagnosis. The methylation status of 3 genes (Ras association domain family 1 isoform A, RASSF1A; opioid-binding protein/cell adhesion molecule, OPCML; homeobox A9, HOXA9) were examined and compared in 35 EOC samples and 11 normal ovarian tissue samples using CCP-based FRET. Gene methylation levels were clustered into 3 sections and assigned a value; values for the 3 genes were summed. Although methylation of the OPCML gene was significantly associated with stage, histological types, grade, and ascites and that of RASSF1A and HOXA9 was not, the sum for the 3 genes was significantly associated with stage and ascites. The sum showed higher sensitivity (85.7%) and specificity (100%) for discriminating EOC from normal ovarian tissues than did the methylation status of RASSF1A, OPCML, and HOXA9 (48.6, 77.1, 77.1, and 100, 88.1, 100%, respectively). Combining cancer antigen 125 levels with the sum increased the sensitivity to 94.3%. The detection and analysis of a panel of genes' methylation status with the CCP-based FRET technique may be useful for diagnosis and screening of EOC; the associated cancer antigen 125 can be used to increase diagnostic sensitivity.

Expression of p53 upregulated modulator of apoptosis (PUMA) and C-myb in gallbladder adenocarcinoma and their pathological significance.

PURPOSE: An increasing number of studies have shown that PUMA and C-myb signaling pathways are involved in various human cancers including colon carcinomas. However, few studies have examined gallbladder cancer specimens, and little is known about the clinical and pathological significance signaling changes may have in gallbladder adenocarcinoma. This study has investigated the expression of PUMA and C-myb in benign and malignant lesions of gallbladder and its pathological significance. METHODS: Tissue specimens from 108 gallbladder adenocarcinoma patients, 46 adjacent tissues, 15 cases of adenomatous polyps, and 35 surgical specimens from chronic cholecystitis patients were routinely paraffin embedded and sectioned. PUMA and C-myb expressions were detected with EnVision immunohistochemistry. RESULTS: Positive rates of PUMA and C-myb are significantly higher in gallbladder adenocarcinoma tissues than that in the other three (P < 0.01). Gallbladder epithelial cells in PUMA and/or C-myb positive benign cases manifest moderate to severe atypical dysplasia. Positive rates of PUMA and C-myb in well-differentiated tumors with maximum diameter of <2 cm and with no lymph node metastasis and invasion of the surrounding tissues are significantly lower than that in those poorly differentiated cases with maximum diameter of ≥ 2 cm, lymph node metastasis, and invasion of the surrounding tissues (P < 0.05 or P < 0.01). The postoperative survival of patients whose tumor specimens are positive for PUMA and C-myb is significantly shorter than that of those who are negative for both markers (P < 0.05 or P < 0.01). CONCLUSIONS: Our results have demonstrated that PUMA and C-myb positive gallbladder tumors progress rapidly, are prone to metastasis, possess strong invasive ability, and have poor prognosis.

Hematopoietic stem cell transplantation for autoimmune diseases in developing countries: current status and future prospectives.

In this paper we present preliminary results of hematopoietic stem cell transplantation for autoimmune diseases in Brazil and China. Chinese experience transplanting lupus is significant and the Brazilian experience with several autoimmune diseases is growing. We discuss peculiar conditions in developing countries which could affect the results, and future prospectives for the organization of phase III randomized trials in those countries.