诱饵受体3及肿瘤坏死因子配体相关分子1A在CD_(4)和CD_(8)双阴性T细胞治疗裸鼠胰腺癌种植瘤中的作用

Role of decoy receptor 3 and its tumor necrosis factor ligand-related molecular 1A in the treatment of pancreatic cancer implantation with CD_(4)and CD_(8)double negative T cells in nude mice

目的:探讨诱饵受体3(DcR3)及其配体肿瘤坏死因子配体相关分子1A(TL1A)在CD_(4)和CD_(8)双阴性T细胞(DNT细胞)治疗裸鼠胰腺癌种植瘤中的作用。方法:采用抗体吸附法体外培养健康志愿者外周血来源的DNT细胞。建立裸鼠胰腺癌种植瘤模型,随机分为DNT细胞治疗组(尾静脉注射1×10^(8)/ml DNT细胞悬液)、吉西他滨治疗组(尾静脉注射50 mg/kg吉西他滨)和对照组(不做任何处理),50 d后测量各组裸鼠种植瘤体积和重量。采用免疫蛋白印迹法和qRT-PCR法检测各组裸鼠种植瘤组织DcR3、TL1A蛋白及mRNA的表达,采用TUNEL法检测各组裸鼠种植瘤的细胞凋亡率。结果:DNT细胞治疗组、吉西他滨治疗组、对照组种植瘤体积分别为(670.28±124.54)、(604.60±179.16)、(1738.80±391.39)mm^(3),瘤体重量分别为(225.60±8.12)、(222.69±8.73)、(265.07±10.76)mg,DNT细胞治疗组、吉西他滨治疗组种植瘤体积和重量显著低于对照组,差异均有统计学意义(P值均<0.001)。DNT细胞治疗组、吉西他滨治疗组DcR3蛋白和mRNA的表达量(0.56±0.02、3.74±0.19,0.57±0.03、3.40±0.39)显著高于对照组(0.39±0.04、0.92±0.05),而TL1A蛋白和mRNA的表达量(0.41±0.03、0.83±0.11,0.40±0.05、0.79±0.08)显著低于对照组(0.81±0.05、1.70±0.36),差异均有统计学意义(P值均<0.001)。DNT细胞治疗组种植瘤的细胞凋亡率为(53.2±11.2)%,吉西他滨治疗组为(56.2±8.6)%,均显著高于对照组的(10.3±3.2)%,差异均有统计学意义(P值均<0.001)。结论:DNT细胞对裸鼠胰腺癌种植瘤生长具有明显的抑制效果,DcR3-TL1A可能参与了DNT细胞的抑癌作用机制。

Objective To explore the role of decoy receptor 3(DcR3)and its ligand tumor necrosis factor ligand-related molecule 1A(TL1A)in the treatment of pancreatic cancer implanted tumors in nude mice with CD4 and CD8 double negative T cells(DNT cells).Methods DNT cells derived from peripheral blood of healthy volunteers were cultured in vitro by antibody adsorption method.A nude mouse model of pancreatic cancer implantation was established and randomly divided into DNT cell treatment group(tail vein injection of 1×10^(8)/ml DNT cell suspension),gemcitabine treatment group(tail vein injection of 50 mg/kg gemcitabine)and control group(no treatment).The tumor volume and weight in each group were measured after 50 days.Western blotting and qRT-PCR were used to detect the protein and mRNA expression of DcR3 and TL1A in nude mice implanted tumor tissues in each group,and TUNEL method was used to detect the apoptosis rate of nude mice implanted tumors in each group.Results The tumor volume of the DNT cell treatment group,gemcitabine treatment group,and control group was(670.28±124.54),(604.60±179.16),(1738.80±391.39)mm^(3),and the tumor weight was(225.60±8.12),(222.69±8.73),(265.07±10.76)mg,and the volume and weight of implanted tumors in the DNT cell treatment group and gemcitabine treatment group were significantly lower than those in the control group,and the differences were statistically significant(all P values<0.001).The expression levels of DcR3 protein and mRNA in the DNT cell treatment group and gemcitabine treatment group(0.56±0.02,3.74±0.19;0.57±0.03,3.40±0.39)were significantly higher than those in the control group(0.39±0.04,0.92±0.05),while the expression levels of TL1A protein and mRNA(0.41±0.03,0.83±0.11;0.40±0.05,0.79±0.08)were significantly lower than those of the control group(0.81±0.05,1.70±0.36),and the differences were statistically significant(all P values<0.001).The apoptotic rate of implanted tumors in the DNT cell treatment group was(53.2±11.2)%,and that in the gemcitabine treatment group was(56.2±8.6)%,which were significantly higher than the control group(10.3±3.2)%,and the differences were statistically significant(all P values<0.001).Conclusions DNT cells had a significant inhibitory effect on the growth of pancreatic cancer implanted tumors in nude mice.DcR3-TL1A may be involved in the anti-tumor mechanism of DNT cells.