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

• Published in: Xi 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
• Language: Chinese
• Published: China 01.01.2013
• Subjects: 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
• ISSN: 1007-8738

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