[Adeno-associated virus mediated T-bet gene transfer into SGC-7901 cell to regulate IFN-gamma production].
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.
Qiu
Gufeng
G
Department of Immunology, School of Medical Technology, Jiangsu University, Zhenjiang 212013, China.
Wang
Suoying
S
Wang
Shengjun
S
Shao
Qixiang
Q
Ma
Jie
J
Yang
Ming
M
Xu
Xiaopeng
X
Mao
Chaoming
C
Su
Zhaoliang
Z
Huang
Xinxiang
X
Xu
Huaxi
H
D045744:Cell Line, Tumor; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D013274:Stomach Neoplasms / Q000235:genetics* / Q000378:metabolism; D020825:T-Box Domain Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D000098815:T-bet Transcription Factor
D011994:Recombinant Proteins; D020825:T-Box Domain Proteins; D000098815:T-bet Transcription Factor; D049452:Green Fluorescent Proteins; D007371:Interferon-gamma