Silencing Itch gene expression by ultrasound-targeted microbubble destruction to enhance the immune killing efficiency of T lymphocyte against lung neoplasms cells LA795 / 基础医学与临床

ABSTRACT

Objective To silence the expression of Itch gene of T-lymphocytes by ultrasound-targeted microbubble destruction (UTMD) and to investigate the cytotoxicity of transfected T lymphocytes against LA795 lung neoplasms cells in vitro. Methods T lymphocyte were isolated by magnetic bead,and an targeted shRNA to silence Itch gene of T lymphocytes was established.48 hours after transfection by UTMD,the transfection efficiency was detected and analyzed by fluorescence microscopy and flow cytometry;the expression of Itch protein was measured with Western blot;72 hours after transfection,the secretion of IL-2 and IFN-γ in the cell supernatant was measured by enzyme-linked immunosorbent assay (ELISA). The cytotoxicity activity changes against LA795 lung neoplasms cells was compared between transfected T lymphocytes and negative control or simplex T lymphocytes in vitro. Results The transfection rate to silence Itch gene of T lymphocytes by UTMD was 52.3%±3.8%. At 48 h after transfection,the Itch gene expression can be effectively suppressed by UTMD. Seventy-two hours after transfection,the secretion level of cytokines including IL-2 and IFN-γ,was significantly increased in the group of targeted shRNA to silence Itch gene of T lymphocytes by UTMD(P<0.05) and transfected T lymphocyte also showed more efficient killing ability against LA795 lung neoplasms cells than negative control or blank group at E : T of 10 : 1,20 : 1 and 40 : 1 (P<0.05). Conclusions Silencing the expression of Itch by UTMD can significantly promote immune activity of T lymphocyte,enhance the cytotoxicity activity of T lymphocyte against LA795 lung neoplasms cells in vitro.