ABSTRACT
<p><b>OBJECTIVE</b>To investigate the inhibitory activities of norcantharidin (NCTD), a demethylated analogue of cantharidin, on Hep3B cells (a human hepatoma cell line) with deficiency of p53.</p><p><b>METHODS</b>The survival rate of the Hep3B cells after treating with NCTD was measured by MTT assay. Cell cycle of treated cells was analyzed by flow cytometry, and DNA fragmentation was observed by electrophoresis. The influence of inhibitors for various caspases and anti-death receptors antibodies on the NCTD-induced apoptosis in the cells was determined.</p><p><b>RESULTS</b>NCTD treatment resulted in growth inhibition of Hep3B cells in a dose- and time-dependent manner. Cell cycle analysis of the cells after treatment with NCTD for 48 h shows that NCTD induced G(2)M phase arrest occurs at low concentration ([Symbol: see text] 25 μmol/L) but G(0)G(1) phase arrest at high concentration (50 μmol/L). The addition of both caspase-3 and caspase-10 inhibitors completely inhibited DNA fragmentation. Addition of anti-TRAIL/DR5 antibody significantly inhibited DNA fragmentation.</p><p><b>CONCLUSION</b>NCTD may inhibit the proliferation of Hep3B cells by arresting cell cycle at G(2)M or G(0)G(1) phase, and induce cells apoptosis via TRAIL/DR5 signal transduction through activation of caspase-3 and caspase-10 by a p53-independent pathway.</p>
Subject(s)
Humans , Antibodies, Neoplasm , Pharmacology , Antibodies, Neutralizing , Pharmacology , Apoptosis , Bridged Bicyclo Compounds, Heterocyclic , Pharmacology , Carcinoma, Hepatocellular , Pathology , Caspase 10 , Metabolism , Caspase 3 , Metabolism , Caspase Inhibitors , Pharmacology , Cell Cycle Checkpoints , Cell Line, Tumor , Cell Proliferation , DNA Fragmentation , Immunohistochemistry , Liver Neoplasms , Pathology , Receptors, TNF-Related Apoptosis-Inducing Ligand , Metabolism , Signal Transduction , TNF-Related Apoptosis-Inducing Ligand , Metabolism , Tumor Suppressor Protein p53 , MetabolismABSTRACT
The mitogen-activated protein kinase (MAPK) phosphatase-1 (MKP-1) belongs to the MAPK cascades which are central to cell proliferation and apoptosis. The carcinogenic role of MKP-1 has been reported in many types of cancer but it has rarely been investigated in breast cancer. The present study was designed to evaluate the MKP-1 mRNA expression and its possible regulation by methylation of MKP-1 promoter in the model of several breast cancer cell lines and tissues as well as controls. Our data demonstrate MKP-1 mRNA expression significantly decreased in five breast cancer cell lines compared to breast controls (P < 0.01). Using the methylation-specific PCR (MSP) analysis, the unmethylated reaction (U) is dominant in both normal cell lines and benign breast tumors (100% vs. 86.2%), whereas the methylated reaction (M) is dominant in both breast cancer cell lines and invasive breast tumors (100% vs. 57.2%). In terms of methylation ratio (M/M+U), methylation level in MKP-1 promoter is significantly higher in the invasive breast tumor tissues (n = 152) than in benign breast tumor tissues (n = 29) (P < 0.0001). Assessing the methylation ratio of the promoter of MKP-1 gene to diagnose the breast malignancy (invasive vs. benign), the area under the receiver-operating characteristic (ROC) curve was 0.809 (95% CI: 0.711-0.906, P < 0.001). The best performance for this prediction has a sensitivity of 76.32% and a specificity of 82.76% at the cutoff value of 0.38. Taken together, we firstly demonstrated that the promoter methylation of MKP-1 gene is a potential breast cancer biomarker for breast malignancy.
Subject(s)
Female , Humans , Breast Neoplasms/diagnosis , Cell Line, Tumor , DNA Methylation/genetics , Dual Specificity Phosphatase 1/genetics , Gene Expression Regulation, Neoplastic , Promoter Regions, Genetic , ROC Curve , Sensitivity and Specificity , Biomarkers, TumorABSTRACT
<p><b>OBJECTIVE</b>To study the value of fine-needle aspiration biopsy (FNAB) in diagnosing primary liver cancer (PLC) and its major complications.</p><p><b>METHODS</b>From June 1, 1985 to May 31, 2005, 2528 patients who were presented with suspected PLC underwent ultrasound-guided FNAB in the Cancer Hospital of Fudan University. The results were retrospectively reviewed.</p><p><b>RESULTS</b>Of those 2528 cases, there was malignancy in 2061 patients (81.53%), of which 1704 were diagnosed as primary liver neoplasms; 41 were diagnosed as metastatic carcinoma, and 316 were not further classified as primary or metastatic. No malignancy was found in 431 cases (17.05%). In 36 cases (1.42%), suspicious malignancy or anaplasia was suggested. Follow-up results showed that all the 2061 positive cases were verified to be malignant and there were no false positive cases. 163 of the 431 negative cases were verified to be malignant in the follow-ups, of which 136 cases were PLC; 28 of the 36 suspicious malignancy or anaplasia were proven to be malignant (all were PLC). The sensitivity, specificity, positive predictive value, negative predictive value and overall accuracy of our FNAB for diagnosing liver malignancy were 91.52%, 100.00%, 100.00%, 59.10% and 92.44%, respectively, and 81.01% cases were diagnosed by FNAB in all the 2096 cases with PLC. Cytological examinations of the smears obtained by FNAB correctly distinguished primary and secondary malignancy in 77.49% of the patients. After FNAB, 11 patients (0.44%) had intraperitoneal hemorrhages and 5 cases (0.20%) had needle tract implantation metastases.</p><p><b>CONCLUSIONS</b>FNAB is important and effective for determining the malignancy potential of liver tumors, especially for PLC. Complications related to FNA were rather rare, therefore, this technique may be easily applied to clinical practice.</p>