The Construction Of IL-33 Lentiviral Vector And The Research On Its Overexpression In CT26 Cells

Objectives: To explore the role of IL-33 in physiopathologic mechanism of colorectal cancer by constructing interleukin 33(IL-33)lentiviral vector and packing it into lentiviral particles to transfect murine colon cancer cell line CT26.Methods: The mIL-33 gene cloned pLV-CMV lentiviral vector containing anti-puromycin,which was used to construct lentiviral vector pLV-CMV-IL-33 with mIL-33.Cotransfected 293 T cells were used to pack virus to collect virus supernatant and transfect CT26 cells after transfection.After 72 h of initial transfection,puromycin was used to screen out stable transfectant.After screening for 2 weeks,RT-PCR,Western blot and ELISA were applied to verify mRNA and protein expressions after disturbance of IL-33 up-regulation.CT26 cells with stable IL-33 up-regulated expression(CT26-IL-33)and the control group(CT26-NC)were constructed.Results: Recombinant lentiviral vector pLV-CMV-IL-33 was constructed successfully and was packed into lentiviral particles.After transfecting CT26 cells,stable high expression of target gene was acquired.Conclusions: Lentiviral vector pLV-CMV-mIL-33 containing mIL-33 gene is constructed successfully.Lentiviral system can mediate target gene to express stably in murine colon cancer cell line CT26 effectively.Virus is packed successfully and infecting CT26 cells effectively also lay a foundation on further exploring the role of IL-33 in cancer physiopathologic mechanism.