Zhengzhou, China
Zhengzhou, China

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Qi Q.,Tianjin Medical University | Qi Q.,Tumor Hospital of Henan | Zhou Q.,Tianjin Medical University | Li Y.,Tumor Hospital of Henan | And 3 more authors.
Chinese Journal of Lung Cancer | Year: 2013

Background and objective Up to know, no any study on using human bone marrow mesenchymal stem cells (hBMSCs) as cells carrier of tumor suppressor gene (IL-24) was reported. The aim of this study is to study the efficiency of transduction of hBMSCs by constructing the lentiviral vector in co-expressing enhanced green fluorescent protein (EGFP) gene and human IL-24 gene, and to lay a foundation for gene therapy of tumor in the future. Methods The lentivector which contain IL-24 and EGFP constructed by recombinant DNA technology were co-transfected to 293FT cells with ViraPowerTM Lentiviral Packaging Mix. The recombinant lentivirus infected with hBMSCs were selected and purified by puromycin. Expression of IL-24 mRNA and IL-24 protein levels were detected by real-time quantitative PCR (qPCR) and ELISA. Results The recombinant lentiviral vector of co-expressing IL-24 gene and EGFP gene were successfully constructed by multisite Gateway technology, virus can be packaged, purified and concentrated successfully, and the virus titer was 7.25×107 PFU/mL. The efficiency of recombinant lentivirus to transduce hBMSCs can reach 100% after selection. The result of qPCR showed that the level of IL-24 mRNA expression in transduced group was significantly higher than that in non-transduced group (P<0.05); ELISA detection confirmed that IL-24 protein expression of transduced group was positive in supernatant and the concentration of IL-24 protein is 40 μg/L, while the non-transduced group was negative. Conclusion Lentiviral vector carrying recombinant IL-24 gene can effectively transduce hBMSCs and express IL-24 protein.

Li Y.,Tumor Hospital of Henan | Jiang D.,Suzhou University | Zhao J.,Suzhou University | Ge J.-F.,Suzhou University | Zheng S.-Y.,Suzhou University
Proceedings - 2010 3rd International Conference on Biomedical Engineering and Informatics, BMEI 2010 | Year: 2010

Lack of surface Fas expression is a main route for apoptotic resistance which is considered an important mechanism of tumorigenesis and tumor progression. Fas and FasL expressions in 110 non-small cell lung carcinomas (NSCLCs) were investigated to evaluate their roles in pulmonary carcinogenesis and to examine the clinico-pathologic significance of Fas expression with its relationship with p53 and bcl-2 over-expressions. Immuno- histochemical analysis using tissue microarray demonstrated that a large proportion of NSCLC patients (60%) showed lack of membranous Fas expression. The Fas- negative cases revealed the significantly lower survival rate than Fas-positive ones. Also, the loss of Fas receptor expression was found more frequently in advanced stage and higher nodal status. FasL protein was increased in most NSCLCs (89%) compared to normal lungs. p53 and bcl-2 over-expressions showed no association with Fas expression. Conclusively, reduced membranous Fas expression as a mechanism of apoptotic resistance is considered to play an important part of the pulmonary carcinogenesis, which may predict poor survival and have a bad prognostic influence. Increased FasL expression is thought to be a basis for the immune evasion in NSCLCs. The rare bcl-2 overexpression suggests that this anti-apoptotic protein is unlikely to play a role in the apoptotic resistance of NSCLCs. ©2010 IEEE.

Li Y.,Tumor Hospital of Henan | Zheng S.-Y.,Suzhou University | Jiang D.,Suzhou University | Ge J.-F.,Suzhou University
Proceedings - 2010 3rd International Conference on Biomedical Engineering and Informatics, BMEI 2010 | Year: 2010

The aim of this study was to investigate the expression of Fas and Fas ligand (FasL) and to determine the signifficance of these molecules in lung cancer cell lines. Immunoblotting, RT- PCR and flow cytometric analyses were carried out to measure the expression of Fas and FasL and to examine their interactions and e.ects on cell growth and apoptosis. Fas and FasL were co-expressed in most of the cell lines but to varying degrees. Apoptosis induced by the agonistic anti-Fas antibody was significantly correlated with Fas expression (P=0.0075), whereas cisplatin-induced apoptosis was not. Upregulation of Fas and FasL expression by the administration of cisplatin was found in 7 of 11 (64%) and 9 of 11 (82%) cell lines,respectively. However, cisplatin-induced apoptosis was not suppressed by antagonistic anti-FasL antibody. Thus, our data indicated that Fas and FasL were co-expressed in lung cancer cell lines, and that Fas ligation induced by agonistic anti-Fas antibody is functional and induced apoptosis that was dependent on the levels of Fas expression. In contrast, Fas-FasL interactions appeared to be non-functional. Furthermore, our results suggest that cisplatin- induced apoptosis in lung cancer cells was independent of the Fas-FasL interaction. ©2010 IEEE.

Zheng S.-Y.,Soochow University of China | Li Y.,Tumor Hospital of Henan | Jiang D.,Soochow University of China | Zhao J.,Soochow University of China | Ge J.-F.,Soochow University of China
Molecular Medicine Reports | Year: 2012

The aim of the present study was to investigate the anticancer effect of quercetin (QC) in the human lung cancer cell line A-549 and further study the mechanism of apoptosis induction by QC. Low differentiation potential A-549 human lung cancer cells were treated with QC at different doses and for different times, and the growth inhibitory rates were detected by MTT assay. Apoptosis induced by QC in A-549 cells was observed by Annexin V/PI double staining and flow cytometric assay. The relative tumor growth ratio of the treated/control tumors (T/C) (%) was chosen to represent the tumor growth inhibition of A-549 cell nude mouse xenografts by QC. Apoptosis of the nude mouse xenografts was observed by Annexin V/PI double staining and flow cytometric assay and DNA fragmentation assay. To further determine the molecular mechanism of apoptosis induced by QC, changes in the expression of bcl-2 and bax genes were detected by RT-PCR. Following incubation with QC, the cell growth of the low differentiation potential A-549 human lung cancer cells was dramatically inhibited in a dose-dependent manner. After the cells were exposed to QC for 24, 48 and 72 h, the IC 50 value was 1.02±0.05, 1.41±0.20 and 1.14±0.19 μmol/l, respectively. Apoptosis in the A-549 cells induced by QC was noted. The apoptotic subpopulation of A-549 cells was approximately 12.96 and 24.58%, respectively, when cells were incubated with 1.2 μmol/l QC for 48 and 72 h. T/C (%) of A-549 nude mouse xenografts was 44.3, when the nude mice were treated with QC (8 mg/kg). Meanwhile, apoptosis induced by QC was observed in the A-549 nude mouse xenografts. Increased expression of the bax gene and decreased expression of the bcl-2 gene were noted using RT-PCR. Our results provide further evidence of the growth inhibition of the A-549 human lung adenocarcinoma cancer cell line by QC. This effect is associated with the induction of apoptosis in A-549 cells and the molecular mechanism may be related to the reduction in expression of the apoptosis-regulating gene bcl-2, and increase in expression of the apoptosis-regulating gene bax. These results were also confirmed in vivo.

Yao Z.,Tumor Hospital of heNan | Liu Y.,Tumor Hospital of heNan | Zhao Y.,Tumor Hospital of heNan | Yao S.,Tumor Hospital of heNan | And 4 more authors.
Chinese Journal of Clinical Oncology | Year: 2011

Objective: To discuss the clinical and pathological characteristics, diagnosis, treatment results, and prognosis of primary pulmonary lymphoma (PPL). Methods: The clinical characteristics, diagnostic approaches, pathologic subtypes, and treatment of 17 PPL cases treated at the Tumor Hospital of He'nan between June 1999 and December 2008 were retrospectively analyzed. The Kaplan-Meier method was used in the survival analysis, and the log-rank method was used in the statistical test. Results: Of the 17 patients, 7 were female and 10 were male, with a median age of 56 years (29-73 years). According to the WHO 2001 lymphoma classification system, all 17 cases of PPL were non-Hodgkin's lymphomas; 13 patients had mucosa-associated lymphoid tissue (MALT) lymphoma. PPL accounted for about 0.81% of malignant lymphomas and 4.74% of primary extranodal lymphomas during the same period at the Tumor Hospital of He'nan. The median follow-up time was 62.5 months (5.2-93 months). The overall survival rate of the 17 patients was 62.5% at 5 years; the 5-year overall survival was superior among patients with MALT lymphoma compared with those with the non-MALT lymphoma (the 5-year overall survival rates were 75% and 50%, respectively; P= 0.047). Conclusion: The incidence of PPL is very low. It is easily misdiagnosed because of its nonspecific clinical and imaging manifestations. Acquiring enough represen-tative tissue specimens for pathologic examination is the key to a definitive diagnosis. At present, there is no therapeutic consensus for these patients. Most of its pathologic subtypes are MALT lymphomas; hence, PPL generally has good prognosis.

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