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Objective To explore the changes of proteomic spectra from plasma of patients with lung cancer or benign lung diseases and health controls in order to establish a primary diagnosis model of lung cancer. Methods The proteomic spectra from plasma of 108 patients with lung cancer, 40 patients with benign lung diseases and 22 healthy individuals were analysed by surface-enhanced laser desorption/ionization time of flight mass spectrometry ( SELDI-TOF-MS). The best decision tree model was established by cluster analysis and principal component analysis. Then the model was blindly validated by the protein of 21 patients with lung benign diseases and 47 patients with stage I lung cancer. Results Twenty-three significantly differentially expressed protein peaks were successfully detected (P <0.001). Blinded validation suggested that the accuracy for diagnosing lung cancer was 72. 06%, the sensitivity and specificity were 72. 34% and 71.43%, respectively, and the positive predictive value and negative predictive value were 85. 0% and 78. 95%, respectively. Conclusion SELDI-TOF-MS protein chip technology provides a new tool for the early diagnosis of lung cancer.
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Objective To explore the effects of inducible co-stimulator (ICOS) gene on the cytotoxic activity of cytokine-induced killer (CIK) cells against cholangiocarcinoma cells. Methods CIK-ICOS cells were obtained by stable transfecting ICOS genes into CIK cells through the adenovirus vector whereas untransfected and EGFP-transfected CIK cells were treated as controls. The proliferation and apoptosis of different CIK cells, as well as their cytotoxicity against cholangiocarcinoma cells in the three groups were detected. The expressions of IFN-T, IL-2 and TNF-α in the supernatant of different CIK cells were measured by ELISA. SCID mice with cholangiocarcinoma were randomly divided into CIK group, CIK-EGFP group, CIK-ICOS group and normal saline group. The cytotoxic activity of CIK-ICOS cells against cholangiocarcinoma cells in vivo was observed. Results CIK-ICOS cells displayed better proliferation than CIK cells and CIK-EGFP cells. At day 20 and 23 of culture, the apoptosis rate of CIK-ICOS cells was 0.69% and 0.89%, respectively, while that of the CIK cells was 2.90% and 4.92%. The cytotoxic effect of CIK-ICOS cells at different E: T ratio against cholangiocarcinoma cells was significantly stronger than that of CIK cells and CIK-EGFP cells (F=13.37, 6.46, 25.51, P<0.05). The concentration of IFN-γ in CIK-ICOS cultured supernatant was (49.50±4.73)μg/L, which was significantly higher than that in the cultured supernatant of CIK cells [(30.53±3.73)μg/L] and CIK-EGFP cells [(30.12±2.64)μg/L](F=38.89, P<0.05). The growth of cholangiocarcinoma was significantly slower in CIK-ICOS group than that in CIK group and CIK-EGFP group, whereas the necrosis area of tumor was larger and the CIK cells in CIK-ICOS group was more than those in the other two groups. Conclusions CIK cells had the function of killing cholangiocarcinoma cells in vitro and in vivo. After ICOS genes were transfected into CIK cells, the survival time of CIK cells in vitro was prolonged and the proliferation of CIK cells was enhanced, as well as the secretion of IFN-γ was increased so that the cytotoxicity of CIK cells against cholangiocarcinoma cells in vitro and in vivo was enhanced.
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BACKGROUND: Vascular endothelial growth factor (VEGF) is able to effectively treat the ischemic heart disease, but in vivo VEGF cannot be maintained effective concentration. OBJECTIVE: To detect the expression of VEGF mRNA and protein in human bone marrow mensenchymal stem cells transferred by VEGF-165 gene. DESIGN, TIME AND SETTING: The empirical study was conducted from March 2006 to April 2007 at Shanghai Chest Hospital. MATERIALS: Human bone marrow mesenchymal stem cell line and VEGF were offered by Basic Laboratory, Thoracic Tumor Institute, Shanghai Chest Hospital. METHODS: hVEGF165 gene was reconstructed in pcPGK-vector and transferred into human bone marrow mensenchymal stem cells (BMSCs) by liposome-mediated method, clone screening by G418. MAIN OUTCOME MEASURES: The mRNA and protein of VEGF gene in transferred cells was detected by reverse transcription-polymerase chain reaction (RT-PCR), Real time PCR, Western Blot, enzyme linked immunosorbent assay (ELISA) in 3 stem cells of pcPGK-VEGF165-IRES-GFP and pcPGK- IRES-GFP, respectively. RESULTS: pcPGK-hVEGF165 vector was reconstructed and transferred into hMSCs successfully. The expression of hVEGF165 in the transfected hMSCs was demonstrated with RT-PCR and Real time PCR. Western Blot and ELISA demonstrated that the expression of hVEGF165 in the transfected hMSCs and VEGF protein in supernatant were significantly more than untransfected hMSCs. CONCLUSION: hVEGF165 can be successfully transfected into BMSCs by using liposome mediated gene transfer. Stably expressed VEGF165 cell line can be obtained..
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Background and purpose:Protein kinase C (PKC) is a potentially important target for cancer therapy due to its potential role in carcinogenesis. Abnormal expression and increasing activity of PKC-?are present in non-small cell lung cancer (NSCLC).The aim of this study is to investigate PKC-? protein expressions in lung cancer tissues by tissue microarray and their significance. Methods:Using tissue microarray, PKC-? protein expression in lung cancer patients was examined by immunohistochemistry. 182 samples of umor tissues and 18 samples of adjacent tissues were taken;of the 182 samples, 160 were NSCLC tissues and 22 were SCLC;102 samples from stage I cases and 58 from stageⅡ-Ⅲ cases.Results:Expression of PKC-? protein was observed in 48.3% (88/182) of cancerous tissues and 16.7% (3/18) of adjacent tissues (P
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<p><b>OBJECTIVE</b>To evaluate the effect of vascular endothelia1 growth factor (VEGF) on the hematogenous metastasis of non-small cell lung cancer (NSCLC).</p><p><b>METHODS</b>The identification of lung cancer cells in the peripheral blood were carried out by cytological, immunohistocytologica1 and immunofluorecent stains respectively, following isolation of cytokeratin-expressing cells with magnetic activated cell sorting. The quantification of cancer cells in the blood was performed according to the established flow cytometric assay. The plasma VEGF was measured by commercially available ELISA kit.</p><p><b>RESULTS</b>The lung cancer cells in the blood, showing a remarkable nuclear polymorphism, expressed the epithelial marker cytokeratin and telomerase reverse transcriptase (hTERT). These cells were stained positive by an NSCLC-specific monoclonal antibody S5Al0-2, but negative by antibodies against CD34 and CD45 antigens. Using the flow cytometric assay, 44 cases (28.6%) of l54 NSCLC patients were found to have cancer cells in their blood, with the incidence of positive cases correlated with the stage of disease. The plasma VEGF level was significantly increased in NSCLC patients in comparison with healthy individuals and patients with benign pulmonary diseases. This level was correlated with the stages of disease in patients with adenocarcinoma. In patiens with cancer cells in their blood, a higher level of plasma VEGF was related with an increased number of cancer cells.</p><p><b>CONCLUSION</b>The plasma VEGF level is increased in NSCLC patients with approximate1y one fourth to have cancer cells in the peripheral blood. In these patients, increased VEGF level promotes hematogenous tumor metastasis, as indicated by a much higher number of cancer cells in the blood.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Carcinoma, Non-Small-Cell Lung , Blood , Pathology , DNA-Binding Proteins , Endothelial Growth Factors , Blood , Enzyme-Linked Immunosorbent Assay , Immunohistochemistry , Keratins , Lung Neoplasms , Blood , Pathology , Lymphokines , Blood , Neoplasm Metastasis , Neoplastic Cells, Circulating , Chemistry , Pathology , Telomerase , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
Purpose:To study the expression of telomerase,p53 and p16 genes in non small cell lung cancer(NSCLC). Methods:The expression of telomerase, p53 and p16 genes were examined in 40 NSCLC cases by SP and TRAP methods, respectively. Results:The positive expression rates of telomerase,p53 and p16 genes in NSCLC were 92.5%, 37.5% and 66.7%, respectively. The positive expression rate of p53 gene correlated closely with the T and pathological staging of NSCLC ( P 0.05). Telomerase activity was correlated with the expression rate of p53 gene and pathological stages but negatively correlated with the ages of NSCLC patients( P
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Purpose:To detect the telomerase activity of lung cancer cells in peripheral blood by TRAP.Methods:The mononuclear cell fraction in peripheral blood was isolated by Ficoll Hypaque gradient centrifugation. Then the telomerase activity of cancer cells in peripheral blood of 49 pre chemotherapy patients was determined by PCR TRAP. Results:In the 49 cases, 65.3%(32/49) cases showed significantly increased telomerase activity, among these cases the telomerase activity was especially high in small cell lung cancer and advanced non small cell lung cancer patients, and telomerase activity in the primary chemotherapy patients was higher than in the recurrent chemotherapy patients. Conclusions:Telomerase activity may be a tumor marker of cancer cells in peripheral blood and can evaluate chemotherapy response in lung cancer.
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Objective To investigate the correlation between the expression of vascular endothelial growth factor-C(VEGF-C) and lymph node metastasis in gastric cancer. Methods The expression of VEGF-C was detected in 30 pairs of fresh primary gastric cancers and their adjacent normal tissues from the same patients by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. Automated image analyzer was used to quantify VEGF-C expression, which was reflected by positive index (PI). Results VEGF-C was highly expressed in gastric cancer cells,whilst its expression was undetectable in adjacent normal tissues. VEGF-C expression was higher in lymph node metastasis group(PI=1.345?0.079) and lymphatic invasive(PI=1.315?0.037) group than that in no-metastasis group(PI=1.156?0.045) and no-invasive group (PI=1.154?0.043)(P