Immune Killing Effect of Silencing TIPE2 Expression on T Lymphocytes Against Lung Adenocarcinoma Cells LA795

Objective To silence the expression of tumor necrosis factor-α-induced protein-8-like 2 (TIPE2) gene of T-lymphocytes by specific small interfering RNA (siRNA) and investigate the cytotoxicity activity of transfected T lymphocytes against lung adenocarcinoma cells LA795 in vitro.

Methods Spleen T lymphocytes of mice were separated by magnetic bead, and we screened and validated an effective siRNA to silence TIPE2 gene of T lymphocytes. After 48h transfection, the secretion of IFN-γ was measured by enzyme-linked immuno-sorbent assay (ELISA). At the end, 72h after transfection, we compared the cytotoxicity activity changes against LA795 cells among transfected T lymphocytes(experimental group), negative control and simplex T lymphocytes(blank group) in vitro.

Results We screened a siRNA sequences to effectively inhibit the expression of TIPE2 gene, the transfection rate was 79.63%, and the inhibition rate for TIPE2 protein expression reached 79%. After 48h transfection, the secretion level of IFN-γ in experimental group, negative control and blank group respectively were (678.96±26.91), (401.69±13.67) and (387.03±15.14)pg/ml (P<0.001). After 72h transfection, compared with negative control and blank group, the transfected T lymphocyte showed more efficient killing ability against LA795 cells in different level of effect/target(20:1, 40:1, 80:1). The killing rate of experimental group reached (47.91±3.25)% in effect/target of 80:1.

Conclusion Silencing TIPE2 could significantly promote the secretion level of IFN-γ of mice T lymphocyte, enhance the cytotoxicity activity of T lymphocyte against LA795 lung adenocarcinoma cells in vitro.