Screening of Highly Expressed CPEΔN Lung Cancer H1299 Cells

• Main Authors: Jing SUN, Guirong ZHANG, Hongyue WANG, Hui SHEN
• Format: Article
• Language: zho
• Published: Chinese Anti-Cancer Association; Chinese Antituberculosis Association 2015-06-01
• Series: Chinese Journal of Lung Cancer
• Subjects: Lung neoplsms; N-terminal truncated carboxypeptidase E; Lentiviral vector
• Online Access: http://dx.doi.org/10.3779/j.issn.1009-3419.2015.06.03

Background and objective The N-terminal truncated carboxypeptidase E (CPEΔN) protein is a novel biomarker of tumor metastasis. This study screened the H1299 cell line with a highly expressed CPEΔN gene for in vivo imaging experiment. Methods Human CPEΔN gene was cloned into the luciferase lentiviral vector. H1299 cells transduced with CPEΔN or control lentiviral vectors were selected with 2 µg/mL puromycin. The expression of CPEΔN was identified through Western blot analysis, and luciferase activity was measured using luciferase reporters. Results The human CPEΔN lentiviral expression vector was successfully constructed. The transfection rate of H1299 cells by the lentivirus achieved 80%, with an infection multiplicity of 20. The H1299 cell line with high CPEΔN (H1299-CPEΔN) expression was established, with an increase in CPEΔN expression by four times compared with the control lentivirus-transfected H1299 cell line (H1299-control). As H1299-CPEΔN and H1299-control can effectively decompose luciferase substrates, they can be applied in in vivo imaging. Conclusion H1299-CPEΔN and H1299-control can be used in in vivo imaging experiment for further research on molecular mechanisms and signal transduction to elucidate the role of CPEΔN in lung cancer metastasis.