食管癌细胞株TE-1经PD-1/PD-L1信号通路抑制共培养T细胞的活化、增殖并促进其凋亡

Oesophageal cancer cell line TE-1 inhibited the activation,proliferation and apoptosis of co-cultured T cells through PD-1/PD-L1 signaling pathway

目的 基于程序性细胞死亡受体蛋白-1/程序性细胞死亡配体蛋白-1(PD-1/PD-L1)信号通路研究食管癌细胞株TE-1抑制共培养T细胞的活化、增殖并促进其凋亡的影响。方法 通过体外细胞转染构建TE-1/PD-L1细胞,TE-1/mock细胞,检测转染后的PD-L1表达判断转染效果。设置4组实验对象:CD8^(+)T细胞+TE-1/PD-L1细胞+IgG抗体、CD8^(+)T细胞+TE-1/mock细胞+IgG抗体、CD8^(+)T细胞+TE-1/PD-L1细胞+PD-L1抗体、CD8^(+)T细胞+TE-1/PD-L1细胞+PD-1抗体。以酶联免疫吸附(ELISA)法比较4组间CD8^(+)T细胞γ-干扰素(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素-10(IL-10)水平;以细胞计数法-8(CCK-8)比较4组间细胞增殖情况;以胱天蛋白酶-3(caspase-3)活性检测试剂盒检测4组间CD8^(+)T细胞的凋亡情况。结果 TE-1组、TE-1/mock组、TE-1/PD-L1组PD-L1的水平分别为4.72±0.68、5.04±0.54和12.21±1.35,表明转染后TE-1/PD-L1细胞的PD-L1表达水平显著升高。CD8^(+)T细胞+TE-1/PD-L1细胞+IgG抗体组、CD8^(+)T细胞+TE-1/mock细胞+IgG抗体组、CD8^(+)T细胞+TE-1/PD-L1细胞+PD-L1抗体组和CD8^(+)T细胞+TE-1/PD-L1细胞+PD-1抗体组上清的IFN-γ水平分别为(107.80±4.97)、(154.00±10.02)、(153.00±9.62)和(145.00±17.28)pg·mL^(-1),TNF-α水平分别为(18.80±2.59)、(36.60±4.51)、(37.00±4.90)和(36.60±3.21)pg·mL^(-1),IL-10水平分别为(47.20±16.99)、(115.00±6.60)、(109.60±5.94)和(116.20±8.79)pg·mL^(-1)。CD8^(+)T细胞+TE-1/PD-L1细胞+IgG抗体组的上述指标与CD8^(+)T细胞+TE-1/mock细胞+IgG抗体组、CD8^(+)T细胞+TE-1/PD-L1细胞+PD-L1抗体组、CD8^(+)T细胞+TE-1/PD-L1细胞+PD-1抗体组比较,差异均有统计学意义(均P<0.05)。结论 食管癌细胞株TE-1通过PD-1/PD-L1信号通路来抑制共培养T细胞的活化、增殖并促进其凋亡。

Objective To explore the effect of esophageal cancer cell line TE-1 on inhibiting the activation,proliferation,and promoting apoptosis of co-cultured T cells based on the PD-1/PD-L1 signaling pathway.Methods Construct TE-1/PD-L1 cells and TE-1/lock cells through in vitro cell transfection,and detect the expression of PD-L1 after transfection to determine the transfection effect.Four groups of experimental subjects were set up:CD8^(+)T cells+TE-1/PD-L1 cells+IgG antibody group,CD8^(+)T cells+TE-1/mock cells+IgG antibody group,CD8^(+)T cells+TE-1/PD-L1 cells+PD-L1 antibody group,and CD8^(+)T cells+TE-1/PD-L1 cells+PD-1 antibody group.The expression levels of γ-interferon(IFN-γ),tumor necrosis factor-α(TNF-α) and interleukin-10(IL-10) secreted by CD8^(+)T cells were compared by enzyme-linked immunosorbent assay(ELISA) among the 4groups;the proliferation of CD8^(+)T cells among the 4 groups was compared by cell counting kit-8(CCK-8);the apoptosis of CD8^(+)T cells was detected by caspase-3 activity detection kit among the 4 groups.Results The levels of PD-L1 in TE-1 group,TE-1/mock group and TE-1/PD-L1 group were 4.72±0.68,5.04±0.54 and12.21±1.35,respectively,indicating a significant increase in PD-L1 expression levels in TE-1/PD-L1 cells after transfection.The expression levels of IFN-γ in the supernatant of CD8^(+)T cells+TE-1/PD-L1 cells+IgG antibody group,CD8^(+)T cells+TE-1/mock cells + IgG antibody group,CD8^(+)T cells+TE-1/PD-L1 cells+PD-L1 antibody group and CD8^(+)T cells+TE-1/PD-L1 cells+PD-1 antibody group were(107.80±4.97),(154.00±10.02),(153.00±9.62) and(145.00±17.28) pg·mL^(-1);the expression levels of TNF-α were(18.80±2.59),(36.60±4.51),(37.00±4.90) and(36.60±3.21) pg·mL^(-1);the expression levels of IL-10were(47.20±16.99),(115.00±6.60),(109.60±5.94) and(116.20±8.79) pg·mL^(-1),respectively.The above indicators of the CD8^(+)T cells+TE-1/PD-L1 cells+IgG antibody group were statistically significant differences from CD8^(+)T cells+TE-1/mock cells+IgG antibody group,CD8^(+)T cells+TE-1/PD-L1 cells+PD-L1 antibody group,and CD8^(+)T cells+TE-1/PD-L1 cells+PD-1 antibody group(all P<0.05).Conclusion The esophageal cancer cell line TE-1 inhibits the activation,proliferation,and promotes apoptosis of co cultured T cells through the PD-1/PD-L1 signaling pathway.