Abstract
Objective The purpose of this study is construct the lentivirus vector and over expressing cell line of human serine/threonine kinase 1 (LKB1) with green fluorescent protein (GFP) tag, and study the normal expression and location of LKB1 protein on osteosarcoma cell U2OS cell line. Methods Human cDNA containing the full-length of LKB1 was used as template for PCR to construct GFP-LKB1 lentivirus vector. The location and expression of LKB1 in mammalian cells were detected by immunofluorescence and Western blotting assay. Results The lentivirus plasmid and over expressing cell line of LKB1 was constructed successfully, and fluorescence localization test data analysis showed that the human LKB1 protein localizes both in cytoplasm and nucleus. Western blotting test show that LKB1 protein can over expression in Eukaryotic cells.CCK-8 detection showed that after 48 hours of transfection, when control group (1.131±0.003) versus LKB1 group (0.739±0.051) that LKB1 group’s activity decreased, and the two groups had a distinct statistical significance. Conclusion In this study, we successfully constructed a cell line overexpressing GFP-LKB1 and LKB1 overexpression significantly inhibited the activity of tumor cells, which provide a foundation for further study on the function and interaction partners of LKB1 protein. Key words: Osteosarcoma; Serine/threonine kinase 1
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
