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THE INFLUENCE OF HUMAN SINGLE CHAIN INTELEUKIN-12 GENE TRANSDUCTION ON THE BIOLOGICAL BEHAVIOR OF HEPATOMA 7721 CELLS 被引量:1
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作者 金莉 来保长 +2 位作者 耿宜萍 王一理 司履生 《Chinese Medical Sciences Journal》 CAS CSCD 2001年第3期147-152,共6页
Objective. To investigate the anti- tumor effects of human single chain interleukin- 12 (hscIL- 12). Method. pcDNA/ hscIL- 12 recombinant was transfected into human hepatic carcinoma cells (7721 cells) by lipofectin m... Objective. To investigate the anti- tumor effects of human single chain interleukin- 12 (hscIL- 12). Method. pcDNA/ hscIL- 12 recombinant was transfected into human hepatic carcinoma cells (7721 cells) by lipofectin method. The 7721/hscIL- 12 cells which secrete hscIL- 12 stably, were obtained via G418 selection, and in vitro the influence of hscIL- 12 gene transduction on the growth of tumor cells was evaluated by cell cycle analysis. In vivo, genetically engineered 7721 cells (7721/hscIL- 12, 7721/pcDNA) and parental cells were implanted into BALB/c nude mice,respectively. 7721/pcDNA and 7721/hscIL- 12 groups were divided into two sub- groups on day 8: one was administered with hPBL twice, 6 days at interval; the other was given equal volume of PBS. Mice were sacrificed on day 26, and spleens and tumors were taken out for histologic assay. Results. hscIL- 12 produced stably by 7721/hscIL- 12 cells had bioactivity, and it was proved by Western blot, immunocytochemistry, and in situ hybridization. In vitro, compared with 7721 and 7721/pcDNA, the 7721/hscIL- 12 grew much more slowly. FACS assay showed apparent G1 arrest of 7721/hscIL- 12 cells. In animal experiment, on day 8 after inoculation, the tumors of 7721 and 7721/pcDNA group were up to 5~ 7mm,while those of 7721/hscIL- 12 group were 2~ 4mm.When treated with hPBL, the tumor of 7721/hscIL- 12 group disappeared completely. Histologically, the tumors from 7721/hscIL- 12 without hPBL treatment had numerous lymphocyte infiltration, the tumor cells displayed depression looking, atrophy, focal necrosis and apoptosis , whereas the tumors of 7721 and 7721/pcDNA groups grew thrivingly. Conclusion. hscIL- 12 transduced 7721 cells could induced significant antitumor immune response which resulted in tumor regression totally when the hPBL was inoculated, and also hscIL- 12 has certain effects on mice immune system. These findings suggest that hscIL- 12 and hscIL- 12 gene therapy might have promising prospects in clinical application. 展开更多
关键词 hscIL- 12 antitumor activity gene transduction IMMUNITY
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Upregulation of drug sensitivity of multidrug resistant SGC7901/VCR human gastric cancer cells by bax gene transduction 被引量:10
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作者 赵燕秋 肖冰 +2 位作者 陈宝军 乔泰东 樊代明 《Chinese Medical Journal》 SCIE CAS CSCD 2000年第11期17-20,共4页
To investigate the role of bax in a vincristine (VCR) induced multidrug resistant (MDR) human gastric cancer cell line, SGC7901/VCR, in which the Bax protein expression level was significantly lower compared with th... To investigate the role of bax in a vincristine (VCR) induced multidrug resistant (MDR) human gastric cancer cell line, SGC7901/VCR, in which the Bax protein expression level was significantly lower compared with that in parent cells Methods A bax eukaryotic expression vector was constructed and transfected into SGC7901/VCR cells by lipofectamine, and resistant clones were selected by G418 Western blotting detected Bax expression in transfectants Tetrazolium blue (MTT) assay evaluated the differences in drug sensitivity and cell cycle changes of transfectants were analyzed using flowcytometry (FCM) Results The bax eukaryotic expression vector was constructed and transfected into SGC7901/VCR cells Through G418 selection, resistant clones were obtained Western blotting demonstrated that the expression of Bax protein was markedly increased in bax transduced cells These cells were more sensitive to adriamycin (ADR) and VCR than mock vector transducted cells Moreover, bax transfection enhanced ADR induced apoptosis and VCR induced G 2/M phase arrest of SGC7901/VCR cells Conclusion Bax was involved in the MDR of SGC7901/VCR cells 展开更多
关键词 bax · gastric cancer · multidrug resistance · gene transduction · apoptoH
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ANTITUMOR EFFECTS OF HUMAN IL-15 GENE MODIFIED LUNG CANCER CELL LINE 被引量:2
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作者 沈永泉 崔莲仙 +2 位作者 何维 薛莉 巴德年 《Chinese Journal of Cancer Research》 SCIE CAS CSCD 1997年第4期8-12,共5页
Human IL15 cDNA fragment, which contains all codons encoding the human IL15 mature protein and signal peptide was transducted into the human lung squmouse cancer cells(PG cells) and murine lung adenocarcinoma cells(LA... Human IL15 cDNA fragment, which contains all codons encoding the human IL15 mature protein and signal peptide was transducted into the human lung squmouse cancer cells(PG cells) and murine lung adenocarcinoma cells(LA795 cell lines). Two IL15 highly expressed cell clones PG1 and LA795A were used to inoculate the nude mice and the T739 syngeneic mice respectively. PG1 cell express higher level of class ⅠMHC molecule on their surface than PG cells. It was shown that the modified LA795A tumor cells grew slowly in T739 mice and induced high levels of CTL/NK/LAK activity in vivo as well, compared with the case of inoculation with LA795 or LA795neo. No significant difference in the tumor growth was observed in groups of the nude mice inoculated by PG1, PG and PGneo cells respectively, except the gene modified cells could not show the lung metastasis of tumors. The supernatants derived from the LA795A cell culture could promote CTL/NK/LAK activity from the whole splenocytes and the CD4/CD8deleted splenic cells in vitro. The results indicated that the IL15 gene transfected tumor cells play important roles in the process of antitumor or antitumor metastasis. 展开更多
关键词 INTERLEUKIN-15 gene therapy TUMOR gene transduction CYTOTOXICITY
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Caspase 3 siRNA decreases apoptosis in cultured neuronal cells
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作者 Chunting Ye Yaoxiong Huang +1 位作者 Xiaohong Yang Honghui Chen 《Neural Regeneration Research》 SCIE CAS CSCD 2009年第9期672-676,共5页
BACKGROUND: Lentiviral vectors, a type of retroviral vector, are able to infect cells at all phases of cell cycle. They are able to express exogenous target genes in vivo over long periods of time with limited immuno... BACKGROUND: Lentiviral vectors, a type of retroviral vector, are able to infect cells at all phases of cell cycle. They are able to express exogenous target genes in vivo over long periods of time with limited immunological reaction. OBJECTIVE: To inhibit neuronal apoptosis by blocking the apoptotic cascade reaction, gene silencing of Caspase 3, and transfection of Caspase 3 short hairpin ribonucleic acid (shRNA) into Neuro 2a cells using a lentiviral vector.DESIGN: TiME-AND SETTING: An observational, genetic engineering cellular biology experiment was performed in Guangzhou Red Cross Hospital and Guangzhou Institute of Traumatic Surgery between March 2007 and June 2008. MATERIALS: PLL3.7, PCMV-VSV-G, and PH'8.9∧PR plasmids were provided by the CBR Institute for Biomedical Research, Harvard Medical School, USA. Staurosporine was purchased from Sigma, USA.METHODS: Caspase 3 siRNA was synthesized and cloned into the PLL3.7 plasmid. The Caspase 3 shRNA-PLL3.7 Ientivirus was generated in 293T cells using a calcium phosphate transfection kit. After the lentivirus was transfected into Neuro 2a cells, apoptosis was induced in both the transfected and untransfected cells by staurosporine. Cell apoptosis was assessed by flow cvtometrv.MAIN OUTCOME MEASURES: Caspase 3 mRNA expression was measured by RT-PCR and Caspase protein expression was assessed by Western blot. Cellular apoptosis was determined by flow cytometry using Annevin V-PE/Taad-Cy7. RESULTS: The transfection rate of caspase 3 shRNA was 〉 98% using the lentiviral vector, RT-PCR and Western blot results demonstrated that significantly reduced Caspase 3 mRNA and protein expression in the transfected Neuro 2a. The control group exhibited 38.7% Annexin V/7aad-positive cells, which suggested apoptotic anaphase, while only 5.0% cells in the Caspase 3 gene silencing group entered apoptotic anaphase. CONCKUSION: Caspase 3 shRNA inhibited Caspase 3 expression in Neuro 2a ceils and decreased drug-induced apoptosis of Neuro 2a cells. 展开更多
关键词 caspase 3 SHRNA gene transduction APOPTOSIS neuronal cell
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OVER-EXPRESSION OF A TRUNCATED TYPE I TGF-P RECEPTOR IN NORMAL DERMAL FIBROBLASTS DECREASES TGF-β1 AUTOPRODUCTION
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作者 刘伟 曹谊林 +2 位作者 蔡泽浩 商庆新 钱云良 《Journal of Shanghai Second Medical University(Foreign Language Edition)》 2003年第1期21-24,共4页
Objective To determine over-expression of a truncated type ⅡTGF-β receptor in down-regulating TGF-β1 auto production in normal dermal fibroblasts. Methods In vitro cultured dermal fibroblasts were treated with rhT... Objective To determine over-expression of a truncated type ⅡTGF-β receptor in down-regulating TGF-β1 auto production in normal dermal fibroblasts. Methods In vitro cultured dermal fibroblasts were treated with rhTGF-β1 (5ng/ml) or recombinant adenovirus containing α truncated type Ⅱ TGF-β receptor gene (50 pfu/cell). Their effects on regulating gene expression of TGF-β1 were observed with Northern Blot. Results rh TGF-β1 up-regulated the gene expression of TGF-β1, (34 %-150%) and type Ⅰ pro-collagen( 13 %- 190%). Overexpression of a truncated receptor Ⅱ decreased the gene expression of TGF-β1 (53%-66%). Conclusion Over-expression of the truncated TGF-β receptor Ⅱdown-regulated TGF-β1 autoproduction via blocking signal transduction of TGF-β. This study may provide a new strategy for scar gene therapy. 展开更多
关键词 TGF-β1 auto production signal transduction gene therapy
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Phage-mediated horizontal gene transfer and its implications for the human gut microbiome
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作者 Tatiana Borodovich Andrey N.Shkoporov +1 位作者 R.Paul Ross Colin Hill 《Gastroenterology Report》 SCIE EI 2022年第1期78-89,共12页
Horizontal gene transfer(HGT)in the microbiome has profound consequences for human health and disease.The spread of antibiotic resistance genes,virulence,and pathogenicity determinants predominantly occurs by way of H... Horizontal gene transfer(HGT)in the microbiome has profound consequences for human health and disease.The spread of antibiotic resistance genes,virulence,and pathogenicity determinants predominantly occurs by way of HGT.Evidence exists of extensive horizontal transfer in the human gut microbiome.Phage transduction is a type of HGT event in which a bacteriophage transfers non-viral DNA from one bacterial host cell to another.The abundance of tailed bacteriophages in the human gut suggests that transduction could act as a significant mode of HGT in the gut microbiome.Here we review in detail the known mechanisms of phage-mediated HGT,namely specialized and generalized transduction,lateral transduction,gene-transfer agents,and molecular piracy,as well as methods used to detect phage-mediated HGT,and discuss its potential implications for the human gut microbiome. 展开更多
关键词 gut phageome horizontal gene transfer gene transduction phage-mediated gene transfer
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