Antiproliferative activity of recombinant human interferon-2 expressed in stably transformed BmN cells

摘要

This study aimed at the generation of a stable transformed silkworm BmN cell line which can continuously express human interferon-λ2 (IFN-λ2) gene, and investigated the antiproliferative activity of this recombinant human IFN-λ2. Silkworm BmN cells were transfected with the recombinant vector pIZT/V5-His harboring the human IFN-λ2 gene. After the BmN cells were transfected with the pIZT/V5-His- hIFN-λ2 vector, the stably transformed BmN cells expressing hIFN-λ2 gene were selected using Zeocin. Following two months of screening, the transformed BmN cell line was obtained.?Stable?transformed BmN cell line?can be maintained at a lower Zeocin concentration. The representing?26 kDa?protein band of?IFN-λ2 was detected by SDS-PAGE and Western blotting.?The expression level of hIFN-λ2, determined by ELISA, was about 8.142 ng?in?4?′?105?cells.?The antiproliferative activity of hIFN-λ2 was determined by MTT assay.?The 50% inhibitory concentrations (IC50) of the recombinant hIFN-λ2?on A549 (lung cancer cells), HL60 (acute promyelocytic leukemia cells), BEL-7402 (liver?cancer cells)?and M231 cells (breast cancer cells) were approximately 3.21, 2.84, 6.29 and 9.32 ng/ml, respectively. In summary, Human?IFN-λ2 can be stably expressed in the transformed BmN cell line, and the expressed recombinant hIFN-λ2 demonstrated antiproliferative activity to tumor cells?in vitro.