Establishment of a pulmonary metastasis model of melanoma with stably expressed hTERT/Luc

To construct the eukaryotic expression vectors of human telomerase reverse transcriptase (hTERT) and luciferase (Luc) gene, and stably transfect B16 cell line with them, and inoculate mice via caudal vein to construct pulmonary metastasis models. The hTERT and Luc cDNA fragments were inserted into e...

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Published inXi bao yu fen zi mian yi xue za zhi Vol. 29; no. 1; p. 89
Main Authors Xiao, Yi, Gao, Jiangping, Gao, Kun, Yan, Jinqi, Zhang, Liang, Wang, Yu, Xu, Yuanji, Wang, Xiaoxiong, Yu, Jiyun
Format Journal Article
LanguageChinese
Published China 01.01.2013
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Abstract To construct the eukaryotic expression vectors of human telomerase reverse transcriptase (hTERT) and luciferase (Luc) gene, and stably transfect B16 cell line with them, and inoculate mice via caudal vein to construct pulmonary metastasis models. The hTERT and Luc cDNA fragments were inserted into eukaryotic expression vectors to obtain the recombinant plasmids pIRES-neo-hTERT and pIRES-hyg3-Luc. The plasmids were then transfected into B16 cells by Lipofectamine(TM); 2000. Afte the stably transfected cell line was screened by G418 and hygromycin B, the transcription and expression of the hTERT and Luc genes were identified by Western blotting and immunofluorescence staining. The stably transfected cells were inoculated via caudal vein into male C56BL/6 mice to construct pulmonary metastasis models. An in vivo imaging system was used to monitor the growth and metastasis of the tumors. A stably transfected cell line was established and the express rates of hTERT and Luc gene in the stable cell line were 84% and
AbstractList To construct the eukaryotic expression vectors of human telomerase reverse transcriptase (hTERT) and luciferase (Luc) gene, and stably transfect B16 cell line with them, and inoculate mice via caudal vein to construct pulmonary metastasis models. The hTERT and Luc cDNA fragments were inserted into eukaryotic expression vectors to obtain the recombinant plasmids pIRES-neo-hTERT and pIRES-hyg3-Luc. The plasmids were then transfected into B16 cells by Lipofectamine(TM); 2000. Afte the stably transfected cell line was screened by G418 and hygromycin B, the transcription and expression of the hTERT and Luc genes were identified by Western blotting and immunofluorescence staining. The stably transfected cells were inoculated via caudal vein into male C56BL/6 mice to construct pulmonary metastasis models. An in vivo imaging system was used to monitor the growth and metastasis of the tumors. A stably transfected cell line was established and the express rates of hTERT and Luc gene in the stable cell line were 84% and
Author Zhang, Liang
Wang, Xiaoxiong
Gao, Jiangping
Yan, Jinqi
Wang, Yu
Yu, Jiyun
Xu, Yuanji
Gao, Kun
Xiao, Yi
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Snippet To construct the eukaryotic expression vectors of human telomerase reverse transcriptase (hTERT) and luciferase (Luc) gene, and stably transfect B16 cell line...
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StartPage 89
SubjectTerms Animals
Cell Line, Tumor
Disease Models, Animal
Gene Expression
Humans
Luciferases - genetics
Lung Neoplasms - genetics
Lung Neoplasms - secondary
Male
Melanoma, Experimental - genetics
Melanoma, Experimental - pathology
Mice
Neoplasm Metastasis
Plasmids - genetics
Telomerase - genetics
Title Establishment of a pulmonary metastasis model of melanoma with stably expressed hTERT/Luc
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Volume 29
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