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[Construction of eukaryotic vector for human interleukin-15 cDNA and its expression in lung carcinoma cell lines]
1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
Objective:
Human interleukin(hIL)-15 expression of lung cancer cell lines transfected by rhIL-15 cDNA in vitro was observed.
Methods:
The recombinant plasmid pL-IL-15-SN was established by inserting IL-15 cDNA into vector pLXSN at sites of EcoR I and BamH I. The pL-IL-15-SN was transfected into human lung squmosuse carcinoma (PG) cell line and murine lung adenocarcinoma (LA795) cell line, respectively. Positive clones were obtained by the selection in G418 conditioned culture. Bioactivity on rIL-15 was detected by dependant-proliferation of CTLL-2 cells in vitro.
Results:
Three positive PG and four positive LA795 cell clones were obtained respectively. The results of the assays for hIL-15 bioactivity showed that the expression levels of rhIL-15 ranged from 142 to 201 or from 138 to 178U/(ml.10(6)) for positive PG cells or LA795 cells, respectively.
Conclusions:
Human and murine lung carcinoma cells transfected with IL-15 cDNA can express hIL-15 with bioactivity.