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 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 98%, respectively. Pulmonary metastasis models were successfully established and the conditions of the lung metastasis could be detected easily using in vivo imaging system. The pulmonary metastasis models with stably expressed hTERT for in vivo imaging system in mice has been successfully established.

Citation

Yi Xiao, Jiangping Gao, Kun Gao, Jinqi Yan, Liang Zhang, Yu Wang, Yuanji Xu, Xiaoxiong Wang, Jiyun Yu. Establishment of a pulmonary metastasis model of melanoma with stably expressed hTERT/Luc]. Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology. 2013 Jan;29(1):89-92

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PMID: 23294724

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