Bioinformatic prediction and validation of cellular-mesenchymal epithelial transition(c-Met)as a target for chimeric antigen receptor T(CAR-T)cell therapy in the treatment of colorectal cancer
Bioinformatic prediction and validation of cellular-mesenchymal epithelial transition(c-Met)as a target for chimeric antigen receptor T(CAR-T)cell therapy in the treatment of colorectal cancer
Objective To explore the potential of the cell surface receptor c-Met as an effective target for chimeric antigen receptor T-cell(CAR-T)therapy in colorectal cancer.Methods The bioinformatics was used to analyze the specific expression of c-Met in colorectal adenocarcinoma(COAD)and its clinical significance.c-Met protein expression was detected by immunohistochemistry in tumor tissues obtained from colorectal cancer patients.Flow cytometry was utilized to assess the expression of c-Met in the HCT116 human colorectal cancer cell line.Additionally,primary T cells isolated from human peripheral blood mononuclear cells(PBMCs)were transduced with a lentivirus to generate second-generation CAR-T cells targeting c-Met,followed by an observation of the inhibitory effects of these c-Met-targeted CAR-T cells on HCT116 cells.Results Immunohistochemistry and bioinformatics data both demonstrated that c-Met was over-expressed in COAD,with patients exhibiting relatively lower expression showing better prognosis.In normal colonic tissue,c-Met was either expressed at low levels or not expressed.Flow cytometry revealed high expression of c-Met in HCT116 cells as well.The c-Met-targeted CAR-T cells were capable of specifically recognizing and targeting antigen-expressing tumor cells.CAR-T cells proliferated specifically under antigenic stimulation,exerting cytotoxic effects on cancer cells and releasing cytokines interleukin 2(IL-2)and interferon-gamma(IFN-γ),thereby demonstrating the biological functions.Conclusion c-Met may be a promising therapeutic target in COAD;c-Met-targeted CAR-T cells demonstrate inhibitory effects on colorectal cancer cells in vitro.
维普
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