1.Adeno-associated virus mediated T-bet gene transfer into SGC-7901 cell to regulate IFN-gamma production.
Gufeng QIU ; Suoying WANG ; Shengjun WANG ; Qixiang SHAO ; Jie MA ; Ming YANG ; Xiaopeng XU ; Chaoming MAO ; Zhaoliang SU ; Xinxiang HUANG ; Huaxi XU
Journal of Biomedical Engineering 2009;26(3):606-619
In order to investigate the effect of T-bet on malignant cells, we selected SGC-7901, a kind of human gastric carcinoma cell line, and used gene clone technique and adeno-associated virus (AAV) packing technology, thus obtaining a recombinant rAAV-eGFP-T-bet and T-bet gene-transfected SGC-7901 cells. Then the function of T-bet gene-infected SGC-7901 cells was researched by detecting the levels of IFN-gamma and T-bet production. The results showed: (1) It was verified that rAAV-T-bet's packing was completed; (2) After SGC-7901 cells was transfected by rAAV-eGFP-T-bet, a green fluorescence was found in about 30%-40% SGC-7901s, and the gene of 1670 bp (T-bet) and 388 bp (IFN-gamma) were generated from SGC-7901s cells; (3) The proteins of IFN-gamma and T-bet secreted by SGC-7901 cells were also detected. These reveal that SGC-7901 cell is efficiently infected by rAAV encoding T-bet, which can induce transfected cells to secret IFN-gamma. It may be useful in the researches on cancer immune therapy of transfecting T-bet gene.
Cell Line, Tumor
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Dependovirus
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genetics
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metabolism
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Green Fluorescent Proteins
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biosynthesis
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Humans
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Interferon-gamma
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biosynthesis
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Recombinant Proteins
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biosynthesis
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genetics
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Stomach Neoplasms
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genetics
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metabolism
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T-Box Domain Proteins
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biosynthesis
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genetics
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Transfection